Effect of pullulan nanoparticle surface charges on HSA complexation and drug release behavior of HSA-bound nanoparticles.
Nanoparticle (NP) compositions such as hydrophobicity and surface charge are vital to determine the presence and amount of human serum albumin (HSA) binding. The HSA binding influences drug release, biocompatibility, biodistribution, and intercellular trafficking of nanoparticles (NPs). Here, we prepared 2 kinds of nanomaterials to investigate HSA binding and evaluated drug release of HSA-bound NPs. Polysaccharides (pullulan) carboxyethylated to provide ionic derivatives were then conjugated to cholesterol groups to obtain cholesterol-modified carboxyethyl pullulan (CHCP). Cholesterol-modified pullulan (CHP) conjugate was synthesized with a similar degree of substitution of cholesterol moiety to CHCP. CHCP formed self-aggregated NPs in aqueous solution with a spherical structure and zeta potential of -19.9 ± 0.23 mV, in contrast to -1.21 ± 0.12 mV of CHP NPs. NPs could quench albumin fluorescence intensity with maximum emission intensity gradually decreasing up to a plateau at 9 to 12 h. Binding constants were 1.12 × 10(5) M(-1) and 0.70 × 10(5) M(-1) to CHP and CHCP, respectively, as determined by Stern-Volmer analysis. The complexation between HSA and NPs was a gradual process driven by hydrophobic force and inhibited by NP surface charge and shell-core structure. HSA conformation was altered by NPs with reduction of α-helical content, depending on interaction time and particle surface charges. These NPs could represent a sustained release carrier for mitoxantrone in vitro, and the bound HSA assisted in enhancing sustained drug release.


Mismodeled purines: implicit alternates and hidden Hoogsteens.
Hoogsteen base pairs are seen in DNA crystal structures, but only rarely. This study tests whether Hoogsteens or other syn purines are either under-modeled or over-modeled, which are known problems for rare conformations. Candidate purines needing a syn/anti 180° flip were identified by diagnostic patterns of difference electron-density peaks. Manual inspection narrowed 105 flip candidates to 20 convincing cases, all at ≤2.7 Å resolution. Rebuilding and refinement confirmed that 14 of these were authentic purine flips. Seven examples are modeled as Watson-Crick base pairs but should be Hoogsteens (commonest at duplex termini), and three had the opposite issue. Syn/anti flips were also needed for some single-stranded purines. Five of the 20 convincing cases arose from an unmodeled alternate duplex running in the opposite direction. These are in semi-palindromic DNA sequences bound by a homodimeric protein and show flipped-purine-like difference peaks at residues where the palindrome is imperfect. This study documents types of incorrect modeling which are worth avoiding. However, the primary conclusions are that such mistakes are infrequent, the bias towards fitting anti purines is very slight, and the occurrence rate of Hoogsteen base pairs in DNA crystal structures remains unchanged from earlier estimates at ∼0.3%.


Deep Neural Network Regression for Automated Retinal Layer Segmentation in Optical Coherence Tomography Images.
Segmenting the retinal layers in optical coherence tomography (OCT) images helps to quantify the layer information in early diagnosis of retinal diseases, which are the main cause of permanent blindness. Thus, the segmentation process plays a critical role in preventing vision impairment. However, because there is a lack of practical automated techniques, expert ophthalmologists still have to manually segment the retinal layers. In this study, we propose an automated segmentation method for OCT images based on a feature-learning regression network without human bias. The proposed deep neural network regression takes the intensity, gradient, and adaptive normalized intensity score (ANIS) of an image segment as features for learning, and then predicts the corresponding retinal boundary pixel. Reformulating the segmentation as a regression problem obviates the need for a huge dataset and reduces the complexity significantly, as shown in the analysis of computational complexity given here. In addition, assisted by ANIS, the method operates robustly on OCT images containing intensity variances, low-contrast regions, speckle noise, and blood vessels, yet remains accurate and time-efficient. In evaluation of the method conducted using 114 images, the processing time was approximately 10.596 s per image for identifying eight boundaries, and the training phase for each boundary line took only 30 s. Further, the Dice similarity coefficient used for assessing accuracy gave a computed value of approximately 0.966. The absolute pixel distance of manual and automatic segmentation using the proposed scheme was 0.612, which is less than a one-pixel difference, on average.


A Season-Long Examination of Team Structure and Its Implications for Subgroups in Individual Sport.
The authors explored how sport structure predisposed a team to subgroup formation and influenced athlete interactions and team functioning. A season-long qualitative case study was undertaken with a nationally ranked Canadian track and field team. Semistructured interviews were conducted with coaches (n = 4) and athletes (n = 11) from different event groups (e.g., sprinters, jumpers) at the beginning and at the end of the season. The results highlighted constraints that directly impacted athlete interactions and predisposed the group to subgroup formation (e.g., sport/event type, facility/schedule limitations, team size/change over time). The constraints led to structural divides that impacted interactions but could be overcome through team building, engaging with leaders, and prioritizing communication. These findings underline how structure imposed by the design of sports impacts teammate interactions and how practitioners, coaches, and athletes can manage groups when facing such constraints. The authors describe theoretical and practical implications while also proposing potential future directions.


Factors Associated With Loss to Follow-up During the First Year After Total Knee Arthroplasty.
Despite increased pressure to capture patient-reported outcome measures for at least 1 year following total joint arthroplasty (TJA), follow-up rates during the first year after TJA are typically lower than desired and may result in biased findings if data are not missing at random. We conducted a retrospective review of medical records of primary total knee arthroplasty patients treated by a single surgeon at an urban academic private hospital. Main measures were demographics (sex, age, race, and insurance), body mass index, travel distance to clinic, and the Knee Injury and Osteoarthritis Outcome Score (KOOS). Multivariable regression analyses were performed to identify patient characteristics associated with attendance at follow-up visits and predictors of attendance at 6-month follow-up. Among the 205 study patients, follow-up visit attendance declined from a high of 95.7% at day 14 to lows of 69.2% at 6 months and 64.4% at 1 year. Attendance at the previously scheduled follow-up visit was a statistically significant predictor of attendance at 3-month (P=.0015), 6-month (P=.0002), and 1-year (P<.0001) follow-up visits, and travel distance was significantly associated with attending the 1-year follow-up visit (P=.042). Patients with the most favorable KOOS Symptom, Pain, and Function in daily living subscale scores at 3-month follow-up were significantly less likely to attend the 6-month follow-up visit than patients with the least favorable KOOS scores. Prospective studies are needed to identify the full range of factors that may contribute to high rates of loss to follow-up after TJA, which should be of concern to researchers, clinicians, and hospitals. [Orthopedics. 2023;46(2):93-97.].


A2B adenosine receptor blockade enhances macrophage-mediated bacterial phagocytosis and improves polymicrobial sepsis survival in mice.
Antimicrobial treatment strategies must improve to reduce the high mortality rates in septic patients. In noninfectious models of acute inflammation, activation of A2B adenosine receptors (A2BR) in extracellular adenosine-rich microenvironments causes immunosuppression. We examined A2BR in antibacterial responses in the cecal ligation and puncture (CLP) model of sepsis. Antagonism of A2BR significantly increased survival, enhanced bacterial phagocytosis, and decreased IL-6 and MIP-2 (a CXC chemokine) levels after CLP in outbred (ICR/CD-1) mice. During the CLP-induced septic response in A2BR knockout mice, hemodynamic parameters were improved compared with wild-type mice in addition to better survival and decreased plasma IL-6 levels. A2BR deficiency resulted in a dramatic 4-log reduction in peritoneal bacteria. The mechanism of these improvements was due to enhanced macrophage phagocytic activity without augmenting neutrophil phagocytosis of bacteria. Following ex vivo LPS stimulation, septic macrophages from A2BR knockout mice had increased IL-6 and TNF-α secretion compared with wild-type mice. A therapeutic intervention with A2BR blockade was studied by using a plasma biomarker to direct therapy to those mice predicted to die. Pharmacological blockade of A2BR even 32 h after the onset of sepsis increased survival by 65% in those mice predicted to die. Thus, even the late treatment with an A2BR antagonist significantly improved survival of mice (ICR/CD-1) that were otherwise determined to die according to plasma IL-6 levels. Our findings of enhanced bacterial clearance and host survival suggest that antagonism of A2BRs offers a therapeutic target to improve macrophage function in a late treatment protocol that improves sepsis survival.


The application of online transcranial random noise stimulation and perceptual learning in the improvement of visual functions in mild myopia.
It has recently been demonstrated how perceptual learning, that is an improvement in a sensory/perceptual task upon practice, can be boosted by concurrent high-frequency transcranial random noise stimulation (tRNS). It has also been shown that perceptual learning can generalize and produce an improvement of visual functions in participants with mild refractive defects. By using three different groups of participants (single-blind study), we tested the efficacy of a short training (8 sessions) using a single Gabor contrast-detection task with concurrent hf-tRNS in comparison with the same training with sham stimulation or hf-tRNS with no concurrent training, in improving visual acuity (VA) and contrast sensitivity (CS) of individuals with uncorrected mild myopia. A short training with a contrast detection task is able to improve VA and CS only if coupled with hf-tRNS, whereas no effect on VA and marginal effects on CS are seen with the sole administration of hf-tRNS. Our results support the idea that, by boosting the rate of perceptual learning via the modulation of neuronal plasticity, hf-tRNS can be successfully used to reduce the duration of the perceptual training and/or to increase its efficacy in producing perceptual learning and generalization to improved VA and CS in individuals with uncorrected mild myopia.


Optical Detection of Intracellular Quantities Using Nanoscale Technologies.
Optical probes that can be used to measure certain quantities with subcellular resolution give us access to a new level of information at which physics, chemistry, life sciences, and medicine become strongly intertwined. The emergence of these new technologies is owed to great advances in the physical sciences. However, evaluating and improving these methods to new standards requires a joint effort with life sciences and clinical practice. In this Account, we give an overview of the probes that have been developed for measuring a few highly relevant parameters at the subcellular scale: temperature, pH, oxygen, free radicals, inorganic ions, genetic material, and biomarkers. Luminescent probes are available in many varieties, which can be used for measuring temperature, pH, and oxygen. Since they are influenced by virtually any metabolic process in the healthy or diseased cell, these quantities are extremely useful to understand intracellular processes. Probes for them can roughly be divided into molecular dyes with a parameter dependent fluorescence or phosphorescence and nanoparticle platforms. Nanoparticle probes can provide enhanced photostability, measurement quality, and potential for multiple functionalities. Embedding into coatings can improve biocompatibility or prevent nonspecific interactions between the probe and the cellular environment. These qualities need to be matched however with good uptake properties, colloidal properties and eventually intracellular targeting to optimize their practical applicability. Inorganic ions constitute a broad class of compounds or elements, some of which play specific roles in signaling, while others are toxic. Their detection is often difficult due to the cross-talk with similar ions, as well as other parameters. The detection of free radicals, DNA, and biomarkers at extremely low levels has significant potential for biomedical applications. Their presence is linked more directly to physiological and clinical manifestations. Since existing methods for free radical detection are generally poor in sensitivity and spatiotemporal resolution, new reliable methods that are generally applicable can contribute greatly to advancing this topic in biology. Optical methods that detect DNA or RNA and protein biomarkers exist for intracellular applications, but are mostly relevant for the development of rapid point-of-care sample testing. To elucidate the inner workings of cells, focused multidisciplinary research is required to define the validity and limitations of a nanoparticle probe, in both physical and biological terms. Multifunctional platforms and those that are easily made compatible with conventional research equipment have an edge over other techniques in growing the body of research evidencing their versatility.


Cardiovascular Reactivity During Marital Conflict in Laboratory and Naturalistic Settings: Differential Associations with Relationship and Individual Functioning Across Contexts.
Cardiovascular reactivity during spousal conflict is considered to be one of the main pathways for relationship distress to impact physical, mental, and relationship health. However, the magnitude of association between cardiovascular reactivity during laboratory marital conflict and relationship functioning is small and inconsistent given the scope of its importance in theoretical models of intimate relationships. This study tests the possibility that cardiovascular data collected in laboratory settings downwardly bias the magnitude of these associations when compared to measures obtained in naturalistic settings. Ambulatory cardiovascular reactivity data were collected from 20 couples during two relationship conflicts in a research laboratory, two planned relationship conflicts at couples' homes, and two spontaneous relationship conflicts during couples' daily lives. Associations between self-report measures of relationship functioning, individual functioning, and cardiovascular reactivity across settings are tested using multilevel models. Cardiovascular reactivity was significantly larger during planned and spontaneous relationship conflicts in naturalistic settings than during planned relationship conflicts in the laboratory. Similarly, associations with relationship and individual functioning variables were statistically significantly larger for cardiovascular data collected in naturalistic settings than the same data collected in the laboratory. Our findings suggest that cardiovascular reactivity during spousal conflict in naturalistic settings is statistically significantly different from that elicited in laboratory settings both in magnitude and in the pattern of associations with a wide range of inter- and intrapersonal variables. These differences in findings across laboratory and naturalistic physiological responses highlight the value of testing physiological phenomena across interaction contexts in romantic relationships.


Introducing a drift and diffusion framework for childhood growth research.
Background: Growth trajectories are highly variable between children, making epidemiological analyses challenging both to the identification of malnutrition interventions at the population level and also risk assessment at individual level. We introduce stochastic differential equation (SDE) models into child growth research. SDEs describe flexible dynamic processes comprising: drift - gradual smooth changes - such as physiology or gut microbiome, and diffusion - sudden perturbations, such as illness or infection. Methods: We present a case study applying SDE models to child growth trajectory data from the Haydom, Tanzania and Venda, South Africa sites within the MAL-ED cohort. These data comprise n=460 children aged 0-24 months. A comparison with classical curve fitting (linear mixed models) is also presented. Results: The SDE models offered a wide range of new flexible shapes and parameterizations compared to classical additive models, with performance as good or better than standard approaches. The predictions from the SDE models suggest distinct longitudinal clusters that form distinct 'streams' hidden by the large between-child variability. Conclusions: Using SDE models to predict future growth trajectories revealed new insights in the observed data, where trajectories appear to cluster together in bands, which may have a future risk assessment application. SDEs offer an attractive approach for child growth modelling and potentially offer new insights.


The comprehensive transcriptional analysis in Caenorhabditis elegans by integrating ChIP-seq and gene expression data.
The fundamental step of learning transcriptional regulation mechanism is to identify the target genes regulated by transcription factors (TFs). Despite numerous target genes identified by chromatin immunoprecipitation followed by high-throughput sequencing technology (ChIP-seq) assays, it is not possible to infer function from binding alone in vivo. This is equally true in one of the best model systems, the nematode Caenorhabditis elegans (C. elegans), where regulation often occurs through diverse TF binding features of transcriptional networks identified in modENCODE. Here, we integrated ten ChIP-seq datasets with genome-wide expression data derived from tiling arrays, involved in six TFs (HLH-1, ELT-3, PQM-1, SKN-1, CEH-14 and LIN-11) with tissue-specific and four TFs (CEH-30, LIN-13, LIN-15B and MEP-1) with broad expression patterns. In common, TF bindings within 3 kb upstream of or within its target gene for these ten studies showed significantly elevated level of expression as opposed to that of non-target controls, indicated that these sites may be more likely to be functional through up-regulating its target genes. Intriguingly, expression of the target genes out of 5 kb upstream of their transcription start site also showed high levels, which was consistent with the results of following network component analysis. Our study has identified similar transcriptional regulation mechanisms of tissue-specific or broad expression TFs in C. elegans using ChIP-seq and gene expression data. It may also provide a novel insight into the mechanism of transcriptional regulation not only for simple organisms but also for more complex species.


Scalable Electronic Ratchet with Over 10% Rectification Efficiency.
Electronic ratchets use a periodic potential with broken inversion symmetry to rectify undirected (electromagnetic, EM) forces and can in principle be a complement to conventional diode-based designs. Unfortunately, ratchet devices reported to date have low or undetermined power conversion efficiencies, hampering applicability. Combining experiments and numerical modeling, field-effect transistor-based ratchets are investigated in which the driving signal is coupled into the accumulation layer via interdigitated finger electrodes that are capacitively coupled to the field effect transistor channel region. The output current-voltage curves of these ratchets can have a fill factor >> 0.25 which is highly favorable for the power output. Experimentally, a maximum power conversion efficiency well over 10% at 5 MHz, which is the highest reported value for an electronic ratchet, is determined. Device simulations indicate this number can be increased further by increasing the device asymmetry. A scaling analysis shows that the frequency range of optimal performance can be scaled to the THz regime, and possibly beyond, while adhering to technologically realistic parameters. Concomitantly, the power output density increases from ≈4 W m-2 to ≈1 MW m-2. Hence, this type of ratchet device can rectify high-frequency EM fields at reasonable efficiencies, potentially paving the way for actual use as energy harvester.


Discovery of an unusual biosynthetic origin for circular proteins in legumes.
Cyclotides are plant-derived proteins that have a unique cyclic cystine knot topology and are remarkably stable. Their natural function is host defense, but they have a diverse range of pharmaceutically important activities, including uterotonic activity and anti-HIV activity, and have also attracted recent interest as templates in drug design. Here we report an unusual biosynthetic origin of a precursor protein of a cyclotide from the butterfly pea, Clitoria ternatea, a representative member of the Fabaceae plant family. Unlike all previously reported cyclotides, the domain corresponding to the mature cyclotide from this Fabaceae plant is embedded within an albumin precursor protein. We confirmed the expression and correct processing of the cyclotide encoded by the Cter M precursor gene transcript following extraction from C. ternatea leaf and sequencing by tandem mass spectrometry. The sequence was verified by direct chemical synthesis and the peptide was found to adopt a classic knotted cyclotide fold as determined by NMR spectroscopy. Seven additional cyclotide sequences were also identified from C. ternatea leaf and flower, five of which were unique. Cter M displayed insecticidal activity against the cotton budworm Helicoverpa armigera and bound to phospholipid membranes, suggesting its activity is modulated by membrane disruption. The Fabaceae is the third largest family of flowering plants and many Fabaceous plants are of huge significance for human nutrition. Knowledge of Fabaceae cyclotide gene transcripts should enable the production of modified cyclotides in crop plants for a variety of agricultural or pharmaceutical applications, including plant-produced designer peptide drugs.


Preventative effect of Zingiber officinale on insulin resistance in a high-fat high-carbohydrate diet-fed rat model and its mechanism of action.
Insulin resistance is a core component of metabolic syndrome and usually precedes the development of type 2 diabetes mellitus. We have examined the preventative effect of an ethanol extract of ginger (Zingiber officinale, Zingiberaceae) on insulin resistance in a high-fat high-carbohydrate (HFHC) diet-fed rat model of metabolic syndrome. The HFHC control rats displayed severe insulin resistance, whilst rats treated with ginger extract (200 mg/kg) during HFHC diet feeding showed a significant improvement of insulin sensitivity using the homeostatic model assessment of insulin resistance (HOMA-IR) after 10 weeks (p < 0.01). An in vitro mechanistic study showed that (S)-[6]-gingerol, the major pungent phenolic principle in ginger, dose-dependently (from 50 to 150 μM) increased AMPK α-subunit phosphorylation in L6 skeletal muscle cells. This was accompanied by a time-dependent marked increment of PGC-1α mRNA expression and mitochondrial content in L6 skeletal muscle cells. These results suggest that the protection from HFHC diet-induced insulin resistance by ginger is likely associated with the increased capacity of energy metabolism by its major active component (S)-[6]-gingerol.


The effects of complex stapes motion on the response of the cochlea.
The piston-like motion of the stapes footplate is the only effective stimulus to the cochlea, and rocking-like stapes motions have no effect on hearing. Studies of the vibration of the stapes in response to acoustic stimulation of the normal ear have revealed a complex movement pattern of its footplate. At low frequencies, the vibrations are predominantly piston-like, but they become increasingly rocking-like at middle and high frequencies. These complex vibrations can be decomposed into a translational, piston-like displacement and 2 rotational movements around the long and short axes of the stapes. The rotational components produce no net volume displacement of the cochlear fluid at some distance from the footplate. Therefore, according to the classic theory of hearing, the rotational motion is not transformed into cochlear activity and a hearing sensation. It was the goal of this study to test this hypothesis experimentally in guinea pigs. A piezoelectric 3-axis device was used to vibrate the stapes in various desired directions while simultaneously monitoring the actual motion of the stapes by a 3-dimensional laser Doppler interferometer and the cochlear activity by recording the compound action potential. The collected data of the presented study cannot be explained by the current theory of hearing. The qualitative results provide supportive evidence that complex movements of the stapes footplate may lead to cochlear activity. Further experiments are necessary to confirm and quantify these effects.


Antioxidant treatment improves neonatal survival and prevents impaired cardiac function at adulthood following neonatal glucocorticoid therapy.
Glucocorticoids are widely used to treat chronic lung disease in premature infants but their longer-term adverse effects on the cardiovascular system raise concerns. We reported that neonatal dexamethasone treatment in rats induced in the short term molecular indices of cardiac oxidative stress and cardiovascular tissue remodelling at weaning, and that neonatal combined antioxidant and dexamethasone treatment was protective at this time. In this study, we investigated whether such effects of neonatal dexamethasone have adverse consequences for NO bioavailability and cardiovascular function at adulthood, and whether neonatal combined antioxidant and dexamethasone treatment is protective in the adult. Newborn rat pups received daily i.p. injections of a human-relevant tapering dose of dexamethasone (D; n = 8; 0.5, 0.3, 0.1 μg g(-1)) or D with vitamins C and E (DCE; n = 8; 200 and 100 mg kg(-1), respectively) on postnatal days 1-3 (P1-3); vitamins were continued from P4 to P6. Controls received equal volumes of vehicle from P1 to P6 (C; n = 8). A fourth group received vitamins alone (CCE; n = 8). At P100, plasma NO metabolites (NOx) was measured and isolated hearts were assessed under both Working and Langendorff preparations. Relative to controls, neonatal dexamethasone therapy increased mortality by 18% (P < 0.05). Surviving D pups at adulthood had lower plasma NOx concentrations (10.6 ± 0.8 vs. 28.0 ± 1.5 μM), an increased relative left ventricular (LV) mass (70 ± 2 vs. 63 ± 1%), enhanced LV end-diastolic pressure (14 ± 2 vs. 8 ± 1 mmHg) and these hearts failed to adapt output with increased preload (cardiac output: 2.9 ± 2.0 vs. 10.6 ± 1.2 ml min(-1)) or afterload (cardiac output: -5.3 ± 2.0 vs.1.4 ± 1.2 ml min(-1)); all P < 0.05. Combined neonatal dexamethasone with antioxidant vitamins improved postnatal survival, restored plasma NOx and protected against cardiac dysfunction at adulthood. In conclusion, neonatal dexamethasone therapy promotes cardiac dysfunction at adulthood. Combined neonatal treatment with antioxidant vitamins is an effective intervention.


Increasing Awareness of the Importance of Physical Activity and Healthy Nutrition: Results From a Mixed-Methods Evaluation of a Workplace Program.
To evaluate the impact of an online workplace program that promotes physical activity and health, while focusing on performance measures relating to physical activity, nutrition, and overall health. The large sample size (more than 18,000 participants) allowed the use of text mining and machine-learning methods to determine what descriptions of the program identify successful outcomes, and hierarchical linear models to determine the most beneficial program modules and features. The program increased overall health and awareness of levels of physical activity and nutrition, especially for people who scored low on these measures initially. Interestingly, although physical activity is the most popular program module, the daily step-tracking process was associated with smaller improvements in overall health. This study finds that the Virgin Pulse Global Challenge is an effective workplace intervention for improving overall health and awareness of physical activity and nutrition. Effectiveness relates to the holistic approach adopted rather than to individual modules in isolation. Future evaluations of workplace health and exercise programs should explore a variety of outcome measures within the rich context provided by open-ended participant experience feedback. In addition, a control group and a follow-up study are required.


(4S,5S)-2,2-Dimethyl-1,3-dioxolane-4,5-dicarbo-nitrile.
The title compound, C7H8N2O2, formed by dehydration of the corresponding dicarboxamide, crystallizes as rectangular prisms. The mol-ecules have a C 2 axis of symmetry through the C atom bearing the methyl groups and the mid-point of the ring C-C bond, and the 1,3-dioxolane ring adopts the extreme twist conformation of the two possible with this symmetry. This brings the two nitrile groups nearest to a linear arrangement when the mol-ecule is viewed along the ring C-C bond. The correct absolute configuration of the mol-ecule was defined by that of the original starting material, (2R,3R)-tartaric acid. The packing is largely controlled by a number of C-H⋯N interactions.


Isolated or non-isolated duodenal obstruction: perinatal outcome following prenatal or postnatal diagnosis.
To determine whether the pre- or postnatal diagnosis of either isolated or non-isolated duodenal obstruction (DO) is associated with different outcomes. A single-center retrospective analysis was carried out of 91 cases diagnosed with a DO between January 1991 and June 2003. Data on the diagnosis, treatment and outcomes of the cases were gathered, and differences between the groups were analyzed. Twenty-eight cases of DO were diagnosed before and 63 after birth. Of 15 presumed isolated cases in the prenatally diagnosed group, four revealed associated or chromosomal anomalies after birth. The types of obstruction present were significantly different between the prenatally (n = 11) and postnatally (n = 27) detected subsets of isolated DO. The prenatally detected subset displayed a lower median gestational age at delivery, lower median birth weight and a higher prematurity rate (8/11 vs. 8/27). The diagnosis of DO occurred significantly later in the postnatally detected subset than the postnatal confirmation of the diagnosis in the prenatally detected cases. In the non-isolated cases of DO, no difference was found in the type of chromosomal or associated anomaly or the type of obstruction between the prenatally detected (n = 17) and postnatally detected subsets (n = 36). Trisomy 21 was present in 7/17 (41%) vs. 22/36 (61%) cases, respectively. Two terminations and three intrauterine deaths occurred in the prenatal non-isolated subset. The liveborn infants from the prenatally detected non-isolated subset (n = 12) showed a significantly higher prematurity rate (9/12 vs.14/36), lower median birth weight and earlier confirmation of diagnosis after delivery. After surgery, outcome was similar between both subsets of isolated and non-isolated DO. All the infants with an isolated DO survived. Neonatal death occurred in three prenatally and five postnatally diagnosed cases with non-isolated DO. The outcome of prenatally and postnatally diagnosed DO is not essentially different despite more prematurity and a lower birth weight in the former. Of the prenatally detected cases of DO assumed to be isolated, 25% revealed additional chromosomal or associated anomalies after delivery, which influenced outcome.


The Shwachman-Bodian-Diamond syndrome protein family is involved in RNA metabolism.
A combination of structural, biochemical, and genetic studies in model organisms was used to infer a cellular role for the human protein (SBDS) responsible for Shwachman-Bodian-Diamond syndrome. The crystal structure of the SBDS homologue in Archaeoglobus fulgidus, AF0491, revealed a three domain protein. The N-terminal domain, which harbors the majority of disease-linked mutations, has a novel three-dimensional fold. The central domain has the common winged helix-turn-helix motif, and the C-terminal domain shares structural homology with known RNA-binding domains. Proteomic analysis of the SBDS sequence homologue in Saccharomyces cerevisiae, YLR022C, revealed an association with over 20 proteins involved in ribosome biosynthesis. NMR structural genomics revealed another yeast protein, YHR087W, to be a structural homologue of the AF0491 N-terminal domain. Sequence analysis confirmed them as distant sequence homologues, therefore related by divergent evolution. Synthetic genetic array analysis of YHR087W revealed genetic interactions with proteins involved in RNA and rRNA processing including Mdm20/Nat3, Nsr1, and Npl3. Our observations, taken together with previous reports, support the conclusion that SBDS and its homologues play a role in RNA metabolism.


Inhibition of peripheral TNF can block the malaise associated with CNS inflammatory diseases.
Circulating cytokine levels are elevated in many neuropathologies and may be a cause of the associated malaise and depression. Using a rat model, we demonstrate that sickness behaviors generated by microinjection of IL-1beta into the anterior hypothalamus are adopted by naive recipient animals following plasma transfer. We further show that neutralizing peripheral TNF by etanercept (a p75 TNF receptor/Fc fusion protein) prior to the IL-1beta microinjection inhibits certain IL-1beta-mediated sickness behaviors, such as the depression of open-field activity and reduced glucose consumption. IL-1beta-induced central lesions induce peripheral TNF as part of the acute-phase response, and this appears to be the principal target of the etanercept. Thus behavioral changes induced by CNS lesions may result from peripheral expression of cytokines that can be targeted with drugs which do not need to cross the blood-brain barrier to be efficacious.


Analyzing large Alzheimer's disease cognitive datasets: Considerations and challenges.
Recent data-sharing initiatives of clinical and preclinical Alzheimer's disease (AD) have led to a growing number of non-clinical researchers analyzing these datasets using modern data-driven computational methods. Cognitive tests are key components of such datasets, representing the principal clinical tool to establish phenotypes and monitor symptomatic progression. Despite the potential of computational analyses in complementing the clinical understanding of AD, the characteristics and multifactorial nature of cognitive tests are often unfamiliar to computational researchers and other non-specialist audiences. This perspective paper outlines core features, idiosyncrasies, and applications of cognitive test data. We report tests commonly featured in data-sharing initiatives, highlight key considerations in their selection and analysis, and provide suggestions to avoid risks of misinterpretation. Ultimately, the greater transparency of cognitive measures will maximize insights offered in AD, particularly regarding understanding the extent and basis of AD phenotypic heterogeneity.


Evaluation of primary HPV-DNA testing in relation to visual inspection methods for cervical cancer screening in rural China: an epidemiologic and cost-effectiveness modelling study.
A new lower-cost rapid-throughput human papillomavirus (HPV) test (careHPV, Qiagen, Gaithersburg, USA) has been shown to have high sensitivity for the detection of high grade cervical intraepithelial neoplasia. We assessed the outcomes and cost-effectiveness of careHPV screening in rural China, compared to visual inspection with acetic acid, when used alone (VIA) or in combination with Lugol's iodine (VIA/VILI). Using data on sexual behaviour, test accuracy, diagnostic practices and costs from studies performed in rural China, we estimated the cost-effectiveness ratio (CER) and associated lifetime outcomes for once-lifetime and twice-lifetime screening strategies, and for routine screening at 5-yearly, 10-yearly and IARC-recommended intervals. The optimal age range for once-lifetime screening was also assessed. For all strategies, the relative ordering of test technologies in reducing cervical cancer incidence and mortality was VIA (least effective); VIA/VILI; firstname.lastname@example.com pg/ml and email@example.com pg/ml (most effective). For once-lifetime strategies, maximum effectiveness was achieved if screening occurred between 35-50 years. Assuming a participation rate of ~70%, once-lifetime screening at age 35 years would reduce cancer mortality by 8% (for VIA) to 12% (for email@example.com) over the long term, with a CER of US$557 (for VIA) to $959 (for firstname.lastname@example.com) per life year saved (LYS) compared to no intervention; referenced to a 2008 GDP per capita in Shanxi Province of $2,975. Correspondingly, regular screening with an age-standardised participation rate of 62% (which has been shown to be achievable in this setting) would reduce cervical cancer mortality by 19-28% (for 10-yearly screening) to 43-54% (using IARC-recommended intervals), with corresponding CERs ranging from $665 (for 10-yearly VIA) to $2,269 (for IARC-recommended intervals using firstname.lastname@example.com) per LYS. This modelled analysis suggests that primary careHPV screening compares favourably to visual inspection screening methodologies in rural China, particularly if used as part of a regular screening program.


Differential effects of IL-21 and IL-15 on perforin expression, lysosomal degranulation, and proliferation in CD8 T cells of patients with human immunodeficiency virus-1 (HIV).
An urgent need exists to devise strategies to augment antiviral immune responses in patients with HIV who are virologically well controlled and immunologically stable on highly active antiretroviral therapy (HAART). The objective of this study was to compare the immunomodulatory effects of the cytokines interleukin (IL)-21 with IL-15 on CD8 T cells in patients with HIV RNA of less than 50 copies/mL and CD4 counts greater than 200 cells/mm.(3) Patient CD8 T cells displayed skewed maturation and decreased perforin expression compared with healthy controls. Culture of freshly isolated patient peripheral-blood mononuclear cells (PBMCs) for 5 hours to 5 days with IL-21 resulted in up-regulation of perforin in CD8 T cells, including memory and effector subsets and virus-specific T cells. IL-21 did not induce T-cell activation or proliferation, nor did it augment T-cell receptor (TCR)-induced degranulation. Treatment of patient PBMCs with IL-15 resulted in induction of perforin in association with lymphocyte proliferation and augmentation of TCR-induced degranulation. Patient CD8 T cells were more responsive to cytokine effects than the cells of healthy volunteers. We conclude that CD8 T cells of patients with HIV can be modulated by IL-21 to increase perforin expression without undergoing overt cellular activation. IL-21 could potentially be useful for its perforin-enhancing properties in anti-HIV immunotherapy.


Bindings of NO, CO, and O2 to multifunctional globin type dehaloperoxidase follow the 'sliding scale rule'.
Dehaloperoxidase-hemoglobin (DHP), a multifunctional globin protein, not only functions as an oxygen carrier as typical globins such as myoglobin and hemoglobin, but also as a peroxidase, a mono- and dioxygenase, peroxygenase, and an oxidase. Kinetics of DHP binding to NO, CO, and O2 were characterized for wild-type DHP A and B and the H55D and H55V DHP A mutants using stopped-flow methods. All three gaseous ligands bind to DHP significantly more weakly than sperm whale myoglobin (SWMb). Both CO and NO bind to DHP in a one-step process to form a stable six-coordinate complex. Multiple-step NO binding is not observed in DHP, which is similar to observations in SWMb, but in contrast with many heme sensor proteins. The weak affinity of DHP for O2 is mainly due to a fast O2 dissociation rate, in accordance with a longer εN-Fe distance between the heme iron and distal histidine in DHP than that in Mb, and an open-distal pocket that permits ligand escape. Binding affinities in DHP show the same 3-4 orders separation between the pairs NO/CO and CO/O2, consistent with the 'sliding scale rule' hypothesis. Strong gaseous ligand discrimination by DHP is very different from that observed in typical peroxidases, which show poor gaseous ligand selectivity, correlating with a neutral proximal imidazole ligand rather than an imidazolate. The present study provides useful insights into the rationale for DHP to function both as mono-oxygenase and oxidase, and is the first example of a globin peroxidase shown to follow the 'sliding scale rule' hypothesis in gaseous ligand discrimination.


Expression and evolution of functionally distinct haemoglobin genes in plants.
Haemoglobin genes have been found in a number of plant species, but the number of genes known has been too small to allow effective evolutionary inferences. We present nine new non-symbiotic haemoglobin sequences from a range of plants, including class 1 haemoglobins from cotton, Citrus and tomato, class 2 haemoglobins from cotton, tomato, sugar beet and canola and two haemoglobins from the non-vascular plants, Marchantia polymorpha (a liverwort) and Physcomitrella patens (a moss). Our molecular phylogenetic analysis of all currently known non-symbiotic haemoglobin genes and a selection of symbiotic haemoglobins have confirmed the existence of two distinct classes of haemoglobin genes in the dicots. It is likely that all dicots have both class 1 and class 2 non-symbiotic haemoglobin genes whereas in monocots we have detected only class 1 genes. The symbiotic haemoglobins from legumes and Casuarina are related to the class 2 non-symbiotic haemoglobins, whilst the symbiotic haemoglobin from Parasponia groups with the class 1 non-symbiotic genes. Probably, there have been two independent recruitments of symbiotic haemoglobins. Although the functions of the two non-symbiotic haemoglobins remain unknown, their patterns of expression within plants suggest different functions. We examined the expression in transgenic plants of the two non-symbiotic haemoglobins from Arabidopsis using promoter fusions to a GUS reporter gene. The Arabidopsis GLB1 and GLB2 genes are likely to be functionally distinct. The class 2 haemoglobin gene (GLB2) is expressed in the roots, leaves and inflorescence and can be induced in young plants by cytokinin treatment in contrast to the class 1 gene (GLB1) which is active in germinating seedlings and can be induced by hypoxia and increased sucrose supply, but not by cytokinin treatment.


On the computational complexity of binary and analog symmetric hopfield nets
We investigate the computational properties of finite binary- and analog-state discrete-time symmetric Hopfield nets. For binary networks, we obtain a simulation of convergent asymmetric networks by symmetric networks with only a linear increase in network size and computation time. Then we analyze the convergence time of Hopfield nets in terms of the length of their bit representations. Here we construct an analog symmetric network whose convergence time exceeds the convergence time of any binary Hopfield net with the same representation length. Further, we prove that the MIN ENERGY problem for analog Hopfield nets is NP-hard and provide a polynomial time approximation algorithm for this problem in the case of binary nets. Finally, we show that symmetric analog nets with an external clock are computationally Turing universal.


Consistency of benefit from an early invasive strategy after fibrinolysis: a patient-level meta-analysis.
Randomised controlled trials have demonstrated improved outcomes with an early invasive strategy compared with routine care after fibrinolysis among patients with ST-elevation myocardial infarction. However, it remains uncertain whether specific patient subsets derive differential benefit from an early invasive strategy. Using patient-level data from seven randomised trials, we studied the relationship between treatment assignment (early invasive vs standard care) and adverse cardiovascular events. The outcomes assessed were death/reinfarction at 30 days and at 1 year, as well as death/reinfarction/recurrent ischaemia, major bleeding and stroke at 30 days. The analyses were conducted in strata (age, sex, diabetes, prior infarction, Killip class, anterior infarction and time from symptom onset to fibrinolysis) to assess for an interaction between the stratifying variable and treatment assigned. There were 101 deaths and 115 recurrent infarctions at 30 days in 3010 patients. There were no strata where an invasive strategy conferred a differential treatment effect. With the exception of a marginally significant interaction between Killip class and treatment for death/reinfarction at 30 days and 1 year (p values for interaction 0.044 and 0.038, respectively), no interactions between the stratifying variables and treatment assignment were observed. Benefit from an early invasive strategy after fibrinolysis for ST-elevation myocardial infarction is similar across patient subgroups stratified by these clinical characteristics. Therefore, prediction of risk and benefit from an early invasive strategy after fibrinolysis for ST-elevation myocardial infarction is best achieved by global risk evaluation rather than specific patient characteristics.


Subspecialization in the human posterior medial cortex.
The posterior medial cortex (PMC) is particularly poorly understood. Its neural activity changes have been related to highly disparate mental processes. We therefore investigated PMC properties with a data-driven exploratory approach. First, we subdivided the PMC by whole-brain coactivation profiles. Second, functional connectivity of the ensuing PMC regions was compared by task-constrained meta-analytic coactivation mapping (MACM) and task-unconstrained resting-state correlations (RSFC). Third, PMC regions were functionally described by forward/reverse functional inference. A precuneal cluster was mostly connected to the intraparietal sulcus, frontal eye fields, and right temporo-parietal junction; associated with attention and motor tasks. A ventral posterior cingulate cortex (PCC) cluster was mostly connected to the ventromedial prefrontal cortex and middle left inferior parietal cortex (IPC); associated with facial appraisal and language tasks. A dorsal PCC cluster was mostly connected to the dorsomedial prefrontal cortex, anterior/posterior IPC, posterior midcingulate cortex, and left dorsolateral prefrontal cortex; associated with delay discounting. A cluster in the retrosplenial cortex was mostly connected to the anterior thalamus and hippocampus. Furthermore, all PMC clusters were congruently coupled with the default mode network according to task-unconstrained but not task-constrained connectivity. We thus identified distinct regions in the PMC and characterized their neural networks and functional implications.


Temporal regulation of murine cytomegalovirus transcription and mapping of viral RNA synthesized at immediate early times after infection.
The replication of murine cytomegalovirus strain Smith in murine embryonic fibroblasts was investigated at immediate early, early, and late times after infection. Cloned subgenomic HindIII fragments of murine cytomegalovirus DNA served to define the regions of transcription. At immediate early times viral RNA classes ranging in size from 5.1 to 1.05 kilobases (kb) were transcribed mainly from the fragments HindIII-K and -L, whereas low levels of transcription were detected from the two termini HindIII-E and HindIII-N. A characteristic pattern of proteins could be translated from immediate early RNA in vitro. At early and late times after infection transcription from all HindIII fragments occurred, but different patterns of transcripts and proteins could be identified. Inhibitors of DNA synthesis induced differences in the late transcription pattern, located in the HindIII-F fragment. The coding region for abundant immediate early transcription could be located at between 0.769 and 0.817 map units. A plasmic clone containing the main part (0.769 to 0.815 map units) of this region was constructed. This region coded for six polyadenylated immediate early RNA species of 5.1, 2.75, 2.0, 1.75, 1.65, and 1.05 kb in size. Only the 1.75-kb RNA originated entirely from the HindIII-L fragment. The 5.1- and 2.75-kb RNA species were encoded by both the HindIII-L and HindIII-K fragments, and the 2.0-, 1.65-, and 1.05-kb RNA species were entirely transcribed within HindIII-K.


Comparison of evolutionary algorithms in gene regulatory network model inference.
The evolution of high throughput technologies that measure gene expression levels has created a data base for inferring GRNs (a process also known as reverse engineering of GRNs). However, the nature of these data has made this process very difficult. At the moment, several methods of discovering qualitative causal relationships between genes with high accuracy from microarray data exist, but large scale quantitative analysis on real biological datasets cannot be performed, to date, as existing approaches are not suitable for real microarray data which are noisy and insufficient. This paper performs an analysis of several existing evolutionary algorithms for quantitative gene regulatory network modelling. The aim is to present the techniques used and offer a comprehensive comparison of approaches, under a common framework. Algorithms are applied to both synthetic and real gene expression data from DNA microarrays, and ability to reproduce biological behaviour, scalability and robustness to noise are assessed and compared. Presented is a comparison framework for assessment of evolutionary algorithms, used to infer gene regulatory networks. Promising methods are identified and a platform for development of appropriate model formalisms is established.


Sympathetic neurons are a powerful driver of myocyte function in cardiovascular disease.
Many therapeutic interventions in disease states of heightened cardiac sympathetic activity are targeted to the myocytes. However, emerging clinical data highlights a dominant role in disease progression by the neurons themselves. Here we describe a novel experimental model of the peripheral neuro-cardiac axis to study the neuron's ability to drive a myocyte cAMP phenotype. We employed a co-culture of neonatal ventricular myocytes and sympathetic stellate neurons from normal (WKY) and pro-hypertensive (SHR) rats that are sympathetically hyper-responsive and measured nicotine evoked cAMP responses in the myocytes using a fourth generation FRET cAMP sensor. We demonstrated the dominant role of neurons in driving the myocyte ß-adrenergic phenotype, where SHR cultures elicited heightened myocyte cAMP responses during neural activation. Moreover, cross-culturing healthy neurons onto diseased myocytes rescued the diseased cAMP response of the myocyte. Conversely, healthy myocytes developed a diseased cAMP response if diseased neurons were introduced. Our results provide evidence for a dominant role played by the neuron in driving the adrenergic phenotype seen in cardiovascular disease. We also highlight the potential of using healthy neurons to turn down the gain of neurotransmission, akin to a smart pre-synaptic ß-blocker.


The planning, implementation and publication of a complex intervention trial for chronic fatigue syndrome: the PACE trial.
The PACE trial was a four-arm trial of specialist medical care, compared with specialist medical care with a supplementary therapy: adaptive pacing therapy, cognitive-behavioural therapy or graded exercise therapy, for patients with chronic fatigue syndrome. The trial found that both cognitive-behavioural and graded exercise therapies were more effective than either of the other two treatments in reducing fatigue and improving physical disability. This paper describes the design, conduct and main results of the trial, along with a description of the challenges that had to be overcome in order to produce clear answers to the clinically important questions the trial posed.


Selecting informative food items for compiling food-frequency questionnaires: comparison of procedures.
The authors automated the selection of foods in a computer system that compiles and processes tailored FFQ. For the selection of food items, several methods are available. The aim of the present study was to compare food lists made by MOM2, which identifies food items with highest between-person variance in intake of the nutrients of interest without taking other items into account, with food lists made by forward regression. The name MOM2 refers to the variance, which is the second moment of the nutrient intake distribution. Food items were selected for the nutrients of interest from 2 d of recorded intake in 3524 adults aged 25-65 years. Food lists by 80 % MOM2 were compared to those by 80 % explained variance for regression on differences between the number and type of food items, and were evaluated on (1) the percentage of explained variance and (2) percentage contribution to population intake computed for the selected items on the food list. MOM2 selected the same food items for Ca, a few more for fat and vitamin C, and a few less for carbohydrates and dietary fibre than forward regression. Food lists by MOM2 based on 80 % of variance in intake covered 75-87 % of explained variance for different nutrients by regression and contributed 53-75 % to total population intake. Concluding, for developing food lists of FFQ, it appears sufficient to select food items based on the contribution to variance in nutrient intake without taking covariance into account.


The Impact of Obesity on Colorectal Surgery: A Survey of Canadian Surgeons.
Over 1.7 billion adults worldwide are considered overweight or obese, with the prevalence of obesity in Canada increasing rapidly. Obesity has been shown to affect surgical outcomes such as local recurrence of cancer and wound infections following colorectal surgery. The objective of this study was to determine the perception/attitudes of Canadian surgeons toward the impact of obesity on the practice of colorectal surgery. A twenty-question survey was administered to Canadian surgeons through mail and email solicited via the Canadian Association of General Surgeons over a period of 2010-2011. The questions focused on surgeon demographics, experience with laparoscopic colon resections and their perception of the impact of obesity toward surgical proficiency and complications. One hundred seventy-seven Canadian surgeons completed the survey. There was a wide range of experience among surgeons in terms of years of practice and number of colon resections performed per year. The majority (72.9%) reported having primary general surgical training. A majority of surgeons (57.7%) identified obesity as a risk factor for colorectal surgery. Furthermore, a majority agreed that obesity is a risk factor for wound infection (97.2%), stomal retraction (90.4%) and stomal herniation (82.5%). While obesity was not considered a contraindication to laparoscopic colon surgery, it was considered to increase operative time (98.3%), cardiovascular (80.2%) and respiratory (95.4%) complications. The majority of surgeons across Canada believe obesity is a risk factor for post-operative complications following laparoscopic colorectal surgery. However, the majority did not consider obesity a contraindication for laparoscopic colon resection. Surgical and peri-operative colorectal protocols may need to be re-assessed to identify methods to manage the obese patient more effectively.


Clinical experience of patients referred to a multidisciplinary cardio-oncology clinic: an observational cohort study.
Cardiovascular disease is the 2nd leading cause of long-term morbidity and mortality in cancer survivors. Cardio-oncology clinics (cocs) have emerged to address the issue; however, there is a paucity of data about the demographics and clinical outcomes of patients seen in the coc setting. Cancer patients referred to The Ottawa Hospital coc were included in this retrospective observational study. Data collected were patient demographics, cancer type and stage, reason for referral, cardiac risk factors, cardiac assessments and treatment, and clinical outcomes. Between 2008 and 2015, 779 patients (516 women, 66%; 263 men, 34%) were referred to the coc. Median age of the patients at cancer diagnosis was 60 years (range: 18-90 years). The most frequent reasons for referral were decreased left ventricular ejection fraction (33%), pre-chemotherapy assessment (14%), and arrhythmia (14%). Treatment with cardiac medication was given in 322 patients (41%), 181 (56%) of whom received more than 2 cardiac medications, with 57 (18%) receiving an angiotensin-converting enzyme inhibitor (acei), 46 (14%) receiving an acei and a beta-blocker, and 38 (12%) receiving a beta-blocker. Of 163 breast cancer patients, 129 (79%) were able to complete targeted therapy with coc co-management. Most of the 779 patients (n = 643, 83%) were alive at the time of the last data collection. This cohort study is one of the largest to report characteristics and clinical outcomes of patients referred to a coc. Collaboration between oncologists and cardiologists resulted in completion of cancer therapy in most patients. Ongoing analysis of referral patterns, management plans, and patient outcomes will help to guide the cardiac care of oncology patients, ultimately optimizing cancer and cardiac outcomes alike.


Hyperscanning of Interactive Juggling: Expertise Influence on Source Level Functional Connectivity.
Hyperscanning studies, wherein brain activity is recorded from multiple participants simultaneously, offer an opportunity to investigate interpersonal dynamics during interactive tasks at the neurophysiological level. In this study, we employed a dyadic juggling paradigm and electroencephalography (EEG) hyperscanning to evaluate functional connectivity between EEG sources within and between jugglers' brains during individual and interactive juggling. We applied graph theoretical measures to identify significant differences in functional connectivity between the individual and interactive juggling conditions. Connectivity was measured in multiple juggler pairs with various skill levels where dyads were either skill-level matched or skill-level unmatched. We observed that global efficiency was reduced during paired juggling for less skilled jugglers and increased for more skilled jugglers. When jugglers were skill-level matched, additional reductions were found in the mean clustering coefficient and small-world topology during interactive juggling. A significant difference in hemispheric brain lateralization was detected between skill-level matched and skill-level unmatched jugglers during interactive juggling: matched jugglers had an increased right hemisphere lateralization while unmatched jugglers had an increased left hemisphere lateralization. These results reveal multiple differences in functional brain networks during individual and interactive juggling and suggest that similarities and disparities in individual skills can impact inter-brain dynamics in the performance and learning of motor tasks.


Low-dose N-acetylcysteine protects rats against endotoxin-mediated oxidative stress, but high-dose increases mortality.
We evaluated the effect of the antioxidant N-acetylcysteine (NAC) on oxidative stress, lung damage, and mortality induced by an endotoxin (lipopolysaccharide, or LPS) in the rat. Continuous intravenous infusion of 275 mg NAC/kg in 48 h, starting 24 h before LPS challenge, decreased hydrogen peroxide (H2O2) concentrations in whole blood (p < 0.01). This decrease was accompanied by fewer histologic abnormalities of the lung and decreased mortality (p < 0.025), compared with rats receiving LPS alone. N-Acetylserine, which has no sulfhydryl group, did not protect rats against LPS toxicity. Improved survival was not associated with an increase in pulmonary reduced glutathione, nor with inhibition of serum tumor necrosis factor (TNF) activity. In vitro, TNF production and DNA binding of nuclear factor kappa B (NF-kappaB) in human Mono Mac 6 cells was only inhibited at concentrations of NAC above 20 mM. High-dose NAC treatment (550 and 950 mg/kg in 48 h) decreased lung GSH (p < 0.05) and resulted in a significantly smaller number of surviving animals when compared with the low-dose NAC group (p < 0.025). In vitro, NAC increased hydroxyl radical generation in a system with Fe(III)-citrate and H2O2 by reducing ferric iron to its catalytic, active Fe2+ form. We conclude that low-dose NAC protects against LPS toxicity by scavenging H2O2, while higher doses may have the opposite effect.


Artificial neural networks: current status in cardiovascular medicine.
Artificial neural networks are a form of artificial computer intelligence that have been the subject of renewed research interest in the last 10 years. Although they have been used extensively for problems in engineering, they have only recently been applied to medical problems, particularly in the fields of radiology, urology, laboratory medicine and cardiology. An artificial neural network is a distributed network of computing elements that is modeled after a biologic neural system and may be implemented as a computer software program. It is capable of identifying relations in input data that are not easily apparent with current common analytic techniques. The functioning artificial neural network's knowledge is built on learning and experience from previous input data. On the basis of this prior knowledge, the artificial neural network can predict relations found in newly presented data sets. In cardiology, artificial neural networks have been successfully applied to problems in the diagnosis and treatment of coronary artery disease and myocardial infarction, in electrocardiographic interpretation and detection of arrhythmias and in image analysis in cardiac radiography and sonography. This report focuses on the current status of artificial neural network technology in cardiovascular medical research.


Prevalence of ultrasound-determined cystic endometrial hyperplasia and the relationship with age in dogs.
To investigate the potential relationship between age and diagnosis of cystic endometrial hyperplasia (CEH) in the bitches, 348 ultrasound examinations from 240 bitches (Labradors, Golden Retrievers, German Shepherds, Flat Coated Retrievers, or crosses of those breeds aged between 1.6 and 7.2 years at examination) were examined. A subpopulation of 32 bitches that had completed their breeding careers at 6 years or more of age was also identified. Of all, 18.3% of the bitches were diagnosed with CEH; these cases were newly diagnosed when bitches were between 2.5 years and 7.3 years of age. The proportion of ultrasound examinations in which CEH was identified increased from 6.8% of examinations on 2-year-old breeding bitches to 60.0% of examinations on 6-year-old bitches. Logistic regression identified a positive correlation between mean age at the examination and presence of CEH (χ(2) = 30.74, degrees of freedom = 1, P < 0.001). For 32 bitches that had completed their breeding career, the prevalence of CEH was 56.3%, age at the diagnosis ranged from 3.8 to 7.3 years, and the proportion of bitches affected with CEH increased from 6.3% at 3 years of age to 56.3% at 7 years of age. These data support the contention that the prevalence of CEH increases with age.


Model Synthesis: A General Procedural Modeling Algorithm.
We present a method for procedurally modeling general complex 3D shapes. Our approach can automatically generate complex models of buildings, man-made structures, or urban data sets in a few minutes based on user-defined inputs. The algorithm attempts to generate complex 3D models that resemble a user-defined input model and satisfy various dimensional, geometric, and algebraic constraints to control the shape. These constraints are used to capture the intent of the user and generate shapes that look more natural. We also describe efficient techniques to handle complex shapes and highlight its performance on many different types of models. We compare model synthesis algorithms with other procedural modeling techniques, discuss the advantages of different approaches, and describe as close connection between model synthesis and context-sensitive grammars.


Prolyl 3-hydroxylase 1 deficiency causes a recessive metabolic bone disorder resembling lethal/severe osteogenesis imperfecta.
A recessive form of severe osteogenesis imperfecta that is not caused by mutations in type I collagen has long been suspected. Mutations in human CRTAP (cartilage-associated protein) causing recessive bone disease have been reported. CRTAP forms a complex with cyclophilin B and prolyl 3-hydroxylase 1, which is encoded by LEPRE1 and hydroxylates one residue in type I collagen, alpha1(I)Pro986. We present the first five cases of a new recessive bone disorder resulting from null LEPRE1 alleles; its phenotype overlaps with lethal/severe osteogenesis imperfecta but has distinctive features. Furthermore, a mutant allele from West Africa, also found in African Americans, occurs in four of five cases. All proband LEPRE1 mutations led to premature termination codons and minimal mRNA and protein. Proband collagen had minimal 3-hydroxylation of alpha1(I)Pro986 but excess lysyl hydroxylation and glycosylation along the collagen helix. Proband collagen secretion was moderately delayed, but total collagen secretion was increased. Prolyl 3-hydroxylase 1 is therefore crucial for bone development and collagen helix formation.


Correlations between perfectionism and coping strategies in response to researcher-selected vignettes or participant-selected events.
Participants, taking classes that met general learning requirements, were 133 men and 240 women from two private, Catholic, liberal arts college campuses. The sample consisted of 131 first-year students, 91 sophomores, 70 juniors, and 81 seniors. The sample was 95.2% Euro-American, consistent with the populations of these campuses, and 98.1% were between 18 and 22 years old. There were five different survey packets. One packet asked participants to report how they coped with a self-selected situation from their own past (Standard Condition). The other four packets asked participants how they would cope with one of four vignettes developed by the researchers. Two vignettes described common social situations, and two described common academic situations a college student would find stressful. The packets were randomly distributed among participants. Several significant correlations between perfectionism and coping strategies were found for the 4 vignettes, but none were found under the Standard Condition. Were there a self-serving bias in participants' reports of past situations in which they remember coping successfully, correlations would be diminished between coping and perfectionism in the Standard Condition.


Intermolecular versus intramolecular interactions of Dictyostelium myosin: possible regulation by heavy chain phosphorylation.
Dictyostelium myosin has been examined under conditions that reveal intramolecular and intermolecular interactions that may be important in the process of assembly and its regulation. Rotary shadowed myosin molecules exhibit primarily two configurations under these conditions: straight parallel dimers and folded monomers. All of the monomers bend in a specific region of the 1860-A-long tail that is 1200 A from the head-tail junction. Molecules in parallel dimers are staggered by 140 A, which is a periodicity in the packing of myosin molecules originally observed in native thick filaments of muscle. The most common region for interaction in the dimers is a segment of the tail about 200-A-long, extending from 900 to 1100 A from the head-tail junction. Parallel dimers form tetramers by way of antiparallel interactions in their tail regions with overlaps in multiples of 140 A. The folded configuration of the myosin molecules is promoted by phosphorylation of the heavy chain by Dictyostelium myosin heavy chain kinase. It appears that the bent monomers are excluded from filaments formed upon addition of salt while the dimeric molecules assemble. These results may provide the structural basis for primary steps in myosin filament assembly and its regulation by heavy chain phosphorylation.


Perceptual decisions regarding object manipulation are selectively impaired in apraxia or when tDCS is applied over the left IPL.
This study evaluated whether apraxia can be understood as due to impaired motor representations or motor imagery necessary for appropriate object-use, imitation, and pantomime. The causal role of the left inferior parietal lobe (IPL), which is heavily implicated in apraxia, is also evaluated. These processes are appraised in light of the proposed ventro-dorsal sub-stream of the classic two visual pathway model, where perceptual information from the ventral stream and the dorsal action stream are integrated and essential for object manipulation. Using a task assessing object-use perception, stroke patients with apraxia demonstrated a selective deficit during perceptual decisions reliant on the integration of visible and known object properties to select the appropriate grasp for object-use. This deficit increased with apraxia severity. A dissociation was evident in these patients showing intact non-motoric perceptual decisions regarding the functional semantic relationship between two objects in the absence of the actor (e.g. how a hammer hits a nail). Converging evidence was found using a modified version of the same task in a neuromodulation study that directly targeted the left IPL in healthy participants using transcranial direct current stimulation (tDCS). Application of inhibitory stimulation over the left IPL reduced performance during perceptual decisions regarding object manipulation whilst performance was unaffected during functional semantic decisions. Excitatory stimulation of the left IPL did not affect performance in either task. Combined, these results suggest that the left inferior parietal lobe is critical for motor imagery, and that apraxia may be caused by an inability to use internal motor representations of object manipulation. These results are discussed in terms of motoric and non-motoric perceptual processes and the proposal of an additional ventro-dorsal sub-stream within the dorsal and ventral visual pathways model.


Disentangling child and family influences on maternal expressed emotion toward children with attention-deficit/hyperactivity disorder.
We used multi-level modelling of sibling-pair data to disentangle the influence of proband-specific and more general family influences on maternal expressed emotion (MEE) toward children and adolescents with attention-deficit/hyperactivity disorder (ADHD). MEE was measured using the Five Minute Speech Sample (FMSS) for 60 sibling pairs (aged 5 through 17 years) each comprising one proband with ADHD and one child without ADHD. Questionnaire measures were used to assess child and adolescent conduct and emotional problems and maternal depression and ADHD. Multi-level models partitioned the effects of five MEE components (initial statement [IS], relationship [REL], warmth [WAR], critical comments [CC], and positive comments [PC]) into proband-specific and general family effects. Significant proband-specific effects were confirmed for all MEE components, with higher levels of MEE expressed toward probands with ADHD than siblings without ADHD. For REL, PC, and CC, this effect was explained by comorbid child conduct problems rather than ADHD. Only low WAR was associated with child ADHD itself. Furthermore, only low WAR was related to variations in more general family characteristics, especially levels of maternal depression. MEE toward children with ADHD was influenced by proband-specific factors. For most components, these were driven by comorbid symptoms of conduct problems rather than ADHD itself. WAR was different; it was influenced by both child-specific and more general characteristics of the family. Further studies utilising a longitudinal design are required to establish the direction of causation and extend our understanding of the relationship between EE components and ADHD.


Patient and Clinician Assessments of Symptomatology Changes on Older Adults Following a Psycho-educational Program for Depression and Anxiety.
Important attention has been given to the assessment of patients' perspectives on treatment, especially as outcomes have been typically evaluated by clinicians. This study examined the association between patient and clinician ratings on perceived improvement and symptomatology changes for an older adult population participating in an ongoing psycho-educational program. Pre-post measures including depression (GDS), anxiety (BAI) and general well-being (GWBS) were collected in a sample of 34 older adults (age = 71.32 ± 6.46 years). Post-testing data included perceived improvement rated by patients, and clinician assessment of depressive symptoms (CS-GDS). Results indicate significant correlations between pre-post changes of the GDS and patients' PIQ (r = -0.37, n = 31, p < 0.05), but not on symptomatic changes of the BAI (r = 0.012, n = 32, p > 0.05) or the GWBS (r = 0.12, n = 31, p > 0.05). Relationships between patients' PIQ and post-ratings on GDS (r = -0.74, n = 33, p < 0.05) and CS-GDS (r = -0.48, n = 32, p < 0.05) are also significant. Results imply that pre-post improvement in depressive symptoms is associated with a patient's perceived improvement and that clinician and patient ratings on depression symptoms post-treatment were both inversely correlated to patients' perceived improvement. Findings suggest that the PIQ is a good indicator to assess symptomatic change by patients and clinicians although they are possibly placing attention on different aspects of treatment outcome, as indicated by differences on sub-scales of the PIQ. Clinicians possibly place a strong focus on assessments of depression symptomatology. Future studies may integrate simultaneous assessments of instruments exploring aspects other than depression, especially those examining representations of illness in older adults.


Vector competence of Aedes albopictus populations for chikungunya virus is shaped by their demographic history.
The mosquito Aedes albopictus is one of the most dangerous invasive species. Its worldwide spread has created health concerns as it is a major vector of arboviruses of public health significance such as chikungunya (CHIKV). Dynamics of different genetic backgrounds and admixture events may have impacted competence for CHIKV in adventive populations. Using microsatellites, we infer the genetic structure of populations across the expansion areas that we then associate with their competence for different CHIKV genotypes. Here we show that the demographic history of Ae. albopictus populations is a consequence of rapid complex patterns of historical lineage diversification and divergence that influenced their competence for CHIKV. The history of adventive populations is associated with CHIKV genotypes in a genotype-by-genotype interaction that impacts their vector competence. Thus, knowledge of the demographic history and vector competence of invasive mosquitoes is pivotal for assessing the risk of arbovirus outbreaks in newly colonized areas.


Reciprocity of social influence.
Humans seek advice, via social interaction, to improve their decisions. While social interaction is often reciprocal, the role of reciprocity in social influence is unknown. Here, we tested the hypothesis that our influence on others affects how much we are influenced by them. Participants first made a visual perceptual estimate and then shared their estimate with an alleged partner. Then, in alternating trials, the participant either revised their decisions or observed how the partner revised theirs. We systematically manipulated the partner's susceptibility to influence from the participant. We show that participants reciprocated influence with their partner by gravitating toward the susceptible (but not insusceptible) partner's opinion. In further experiments, we showed that reciprocity is both a dynamic process and is abolished when people believed that they interacted with a computer. Reciprocal social influence is a signaling medium for human-to-human communication that goes beyond aggregation of evidence for decision improvement.


Optimizing the precision of a multichannel three-polarizer spectroscopic ellipsometer.
We developed a multichannel three-polarizer spectroscopic ellipsometer based on a data acquisition algorithm for achieving optimized precision. This algorithm measures unnormalized Fourier coefficients accurately and precisely. Offset angles for optical elements were obtained as wavelength-independent values using regression calibration. Derived subsets of data reduction functions were used to calculate sample parameters. Correlation coefficients of Fourier coefficients were used to calculate errors in the sample parameters. Mean standard deviations of the sample parameters for each data reduction method were compared to identify the best method. This approach could be used to identify suitable precision optimization methods for other rotating-element ellipsometers.


Oral versus intravenous rehydration therapy in severe gastroenteritis.
A controlled, randomised trial comparing the results of oral rehydration therapy with those of intravenous fluid treatment in 470 children with severe gastroenteritis was undertaken. The oral rehydration therapy was divided into two phases--a rehydration phase that used high sodium isotonic fluid at 40 ml/kg per hour and a maintenance phase using low sodium isotonic fluid (sodium 40, potassium 30, bicarbonate 25, chloride 45, and dextrose 130 mmol/l). The results indicate that oral rehydration treatment, used according to this protocol, is successful in treating severe diarrhoea and dehydration, and has considerable advantages over intravenous fluid therapy in reducing complications associated with the treatment of hypernatraemia, in promoting rapid correction of hypokalaemia and acidosis, in decreasing the duration of diarrhoea, and in promoting a greater weight gain at hospital discharge.


Effect of cytosine arabinoside on viral-specific protein synthesis in cells infected with herpes simplex virus.
The relationship between viral DNA and protein synthesis during herpes simplex virus type 1 (HSV-1) replication in HeLa cells was examined. Treatment of infected cells with cytosine arabinoside (ara-C), which inhibited the synthesis of HSV-1 DNA beyond the level of detection, markedly affected the types and amounts of viral proteins made in the infected cell. Although early HSV-1 proteins were synthesized normally, there was a rapid decline in total viral protein synthesis beginning 3 to 4 h after infection. This is the time that viral DNA synthesis would normally have been initiated. ara-C also prevented the normal shift from early to late viral protein synthesis. Finally, it was shown that the effect of ara-C on late protein synthesis was dependent upon the time after infection that the drug was added. These results suggest that inhibition of progeny viral DNA synthesis by ara-C prevents the "turning on" of late HSV-1 protein synthesis but allows early translation to be "switched off."


What are the respiratory effects of e-cigarettes?
Electronic cigarettes (e-cigarettes) are alternative, non-combustible tobacco products that generate an inhalable aerosol containing nicotine, flavors, propylene glycol, and vegetable glycerin. Vaping is now a multibillion dollar industry that appeals to current smokers, former smokers, and young people who have never smoked. E-cigarettes reached the market without either extensive preclinical toxicology testing or long term safety trials that would be required of conventional therapeutics or medical devices. Their effectiveness as a smoking cessation intervention, their impact at a population level, and whether they are less harmful than combustible tobacco products are highly controversial. Here, we review the evidence on the effects of e-cigarettes on respiratory health. Studies show measurable adverse biologic effects on organ and cellular health in humans, in animals, and in vitro. The effects of e-cigarettes have similarities to and important differences from those of cigarettes. Decades of chronic smoking are needed for development of lung diseases such as lung cancer or chronic obstructive pulmonary disease, so the population effects of e-cigarette use may not be apparent until the middle of this century. We conclude that current knowledge of these effects is insufficient to determine whether the respiratory health effects of e-cigarette are less than those of combustible tobacco products.


Postoperative Liver Failure Risk Score: Identifying Patients with Resectable Perihilar Cholangiocarcinoma Who Can Benefit from Portal Vein Embolization.
Major liver resection for perihilar cholangiocarcinoma (PHC) is associated with a 22% to 33% postoperative liver failure incidence. The aim of this study was analyze the predictive value of future liver remnant (FLR) volume for postoperative liver failure after resection for PHC and to develop a risk score to improve patient selection for portal vein embolization. A consecutive series of 217 patients underwent major liver resection for PHC between 1997 and 2014 at 2 Western centers; FLR volumes were calculated with CT volumetry; other variables included jaundice at presentation, immediate preoperative bilirubin, and preoperative cholangitis. The FLR volume was categorized as <30%, 30% to 45%, or >45%. A risk score for postoperative liver failure (grade B/C according to the International Study Group of Liver Surgery criteria) was developed using multivariable logistic regression with 5 predefined variables. Postoperative liver failure incidence was 24% and liver failure-related mortality was 12%. Risk factors for liver failure were FLR volume <30% (odds ratio 4.2; 95% CI 1.77 to 10.3) and FLR volume 30% to 45% (odds ratio 1.4; 95% CI 10.6 to 3.4). In addition, jaundice at presentation (odds ratio 3.1; 95% CI 1.1 to 9.0), immediate preoperative bilirubin >50 μmol/L (>2.9 mg/dL) (odds ratio 4.3; 95% CI 1.7 to 10.7), and preoperative cholangitis (odds ratio 3.4; 95% CI 1.6 to 7.4) were risk factors for liver failure. These variables were included in a risk score that showed good discrimination (area under the curve 0.79; 95% CI 0.72 to 0.86) and ranking patients in 3 risk sub-groups with predicted liver failure incidence of 4%, 14%, and 44%. The selection of patients for portal vein embolization using only liver volume is insufficient, considering the other predictors of liver failure in PHC patients. The proposed risk score can be used for selection of patients for portal vein embolization, for adequate patient counseling, and identification of other modifiable risk factors besides liver volume.


Development and Assessment of the SMAP Enhanced Passive Soil Moisture Product.
Launched in January 2015, the National Aeronautics and Space Administration (NASA) Soil Moisture Active Passive (SMAP) observatory was designed to provide frequent global mapping of high-resolution soil moisture and freeze-thaw state every two to three days using a radar and a radiometer operating at L-band frequencies. Despite a hardware mishap that rendered the radar inoperable shortly after launch, the radiometer continues to operate nominally, returning more than two years of science data that have helped to improve existing hydrological applications and foster new ones. Beginning in late 2016 the SMAP project launched a suite of new data products with the objective of recovering some high-resolution observation capability loss resulting from the radar malfunction. Among these new data products are the SMAP Enhanced Passive Soil Moisture Product that was released in December 2016, followed by the SMAP/Sentinel-1 Active-Passive Soil Moisture Product in April 2017. This article covers the development and assessment of the SMAP Level 2 Enhanced Passive Soil Moisture Product (L2_SM_P_E). The product distinguishes itself from the current SMAP Level 2 Passive Soil Moisture Product (L2_SM_P) in that the soil moisture retrieval is posted on a 9 km grid instead of a 36 km grid. This is made possible by first applying the Backus-Gilbert optimal interpolation technique to the antenna temperature (TA) data in the original SMAP Level 1B Brightness Temperature Product to take advantage of the overlapped radiometer footprints on orbit. The resulting interpolated TA data then go through various correction/calibration procedures to become the SMAP Level 1C Enhanced Brightness Temperature Product (LiC_TB_E). The LiC_TB_E product, posted on a 9 km grid, is then used as the primary input to the current operational SMAP baseline soil moisture retrieval algorithm to produce L2_SM_P_E as the final output. Images of the new product reveal enhanced visual features that are not apparent in the standard product. Based on in situ data from core validation sites and sparse networks representing different seasons and biomes all over the world, comparisons between L2_SM_P_E and in situ data were performed for the duration of April 1, 2015 - October 30, 2016. It was found that the performance of the enhanced 9 km L2_SM_P_E is equivalent to that of the standard 36 km L2_SM_P, attaining a retrieval uncertainty below 0.040 m3/m3 unbiased root-mean-square error (ubRMSE) and a correlation coefficient above 0.800. This assessment also affirmed that the Single Channel Algorithm using the V-polarized TB channel (SCA-V) delivered the best retrieval performance among the various algorithms implemented for L2_SM_P_E, a result similar to a previous assessment for L2_SM_P.


The couple as context: Latino gay male couples and HIV.
HIV-prevention efforts with gay men in relationships frequently omit primary partners. When they are considered, examinations of race/ethnicity are often overlooked despite higher infection rates among gay men of colour. Acknowledging both the need to contextualise the behaviours that may affect HIV risk for gay men of colour and the disproportionate impact of HIV on Latino gay men, the present study utilised semi-structured, qualitative interviews to explore relationship dynamics, sexual agreements and behaviours, safer sex choices and HIV risk among nine Latino gay male couples. All participants were HIV-negative and in concordant negative relationships. Additionally, nearly all reported closed agreements. Analysis revealed participants engaging in four behaviours that may actively reduce their HIV risk: approaching sexual agreements from a practical standpoint, maintaining a high literacy around HIV, having exposure to social support groups for Latino gay men and finding support in their relationship with another Latino gay man. Additional issues are raised where long-term HIV prevention is concerned. Intimate relationships are an important context for understanding both HIV risk and prevention among Latino gay men. Likewise, race/ethnicity provides an indispensable perspective on all research with gay couples. Future prevention efforts with gay men must strive to include both.


Crystal structure of lac repressor core tetramer and its implications for DNA looping.
The crystal structure of the tryptic core fragment of the lac repressor of Escherichia coli (LacR) complexed with the inducer isopropyl-beta-D-thiogalactoside was determined at 2.6 A resolution. The quaternary structure consists of two dyad-symmetric dimers that are nearly parallel to each other. This structure places all four DNA binding domains of intact LacR on the same side of the tetramer, and results in a deep, V-shaped cleft between the two dimers. Each monomer contributes a carboxyl-terminal helix to an antiparallel four-helix bundle that functions as a tetramerization domain. Some of the side chains whose mutation reduce DNA binding form clusters on a surface near the amino terminus. Placing the structure of the DNA binding domain complexed with operator previously determined by nuclear magnetic resonance onto this surface results in two operators being adjacent and nearly parallel to each other. Structural considerations suggest that the two dimers of LacR may flexibly alter their relative orientation in order to bind to the known varied spacings between two operators.


The Replicative Consequences of Papillomavirus E2 Protein Binding to the Origin Replication Factor ORC2.
The origin recognition complex (ORC) coordinates a series of events that lead to initiation of DNA strand duplication. As a nuclear double stranded DNA plasmid, the papillomavirus (PV) genome resembles a mini-chromosome in infected cells. To initiate its replication, the viral E2 protein binds to and recruits the E1 DNA helicase at the viral origin. PV genome replication program exhibits three stages: initial amplification from a single genome upon infection to a few copies per cell, a cell cycle linked maintenance phase, and a differentiation dependent late stage where the genome is amplified to thousands of copies. Involvement of ORC or other pre-replication complex (pre-RC) factors has not been described. We report that human PV (HPV) and bovine PV (BPV-1) E2 proteins bind to ORC2, however, ORC2 was not detected at the viral origin. Depletion of ORC2 enhanced PV replication in a transient replication model and in keratinocytes stably maintaining viral episomes, while there was no effect on copy number in a cell line with integrated HPV genomes. Consistent with this, occupancy of E1 and E2 at the viral origin increased following ORC2 silencing. These data imply that ORC2 is not necessary for activation of the PV origin by E1 and E2 but instead suppresses E2 replicative function. Furthermore, we observed that over-expression of HPV E2 decreased ORC2 occupation at two known mammalian origins of replication, suggesting that E2 restricts pre-ORC assembly that could otherwise compete for host replication complexes necessary for viral genome amplification. We infer that the ORC2 complex with E2 restricts viral replication in the maintenance phase of the viral replication program and that elevated levels of E2 that occur during the differentiation dependent amplification stage subvert ORC loading and hence DNA synthesis at cellular origins.


Transcriptional responses of Bacillus cereus towards challenges with the polysaccharide chitosan.
The antibacterial activity of the polysaccharide chitosan towards different bacterial species has been extensively documented. The response mechanisms of bacteria exposed to this biopolymer and the exact molecular mechanism of action, however, have hardly been investigated. This paper reports the transcriptome profiling using DNA microarrays of the type-strain of Bacillus cereus (ATCC 14579) exposed to subinhibitory concentrations of two water-soluble chitosan preparations with defined chemical characteristics (molecular weight and degree of acetylation (F(A))). The expression of 104 genes was significantly altered upon chitosan A (weight average molecular weight (M(w)) 36.0 kDa, F(A) = 0.01) exposure and 55 genes when treated with chitosan B (M(w) 28.4 kDa, F(A) = 0.16). Several of these genes are involved in ion transport, especially potassium influx (BC0753-BC0756). Upregulation of a potassium transporting system coincides with previous studies showing a permeabilizing effect on bacterial cells of this polymer with subsequent loss of potassium. Quantitative PCR confirmed the upregulation of the BC0753 gene encoding the K(+)-transporting ATPase subunit A. A markerless gene replacement method was used to construct a mutant strain deficient of genes encoding an ATP-driven K(+) transport system (Kdp) and the KdpD sensor protein. Growth of this mutant strain in potassium limiting conditions and under salt stress did not affect the growth pattern or growth yield compared to the wild-type strain. The necessity of the Kdp system for potassium acquisition in B. cereus is therefore questionable. Genes involved in the metabolism of arginine, proline and other cellular constituents, in addition to genes involved in the gluconeogenesis, were also significantly affected. BC2798 encoding a chitin binding protein was significantly downregulated due to chitosan exposure. This study provides insight into the response mechanisms of B. cereus to chitosan treatment and the significance of the Kdp system in potassium influx under challenging conditions.


Gastrointestinal Adverse Events of Cannabinoid 1 Receptor Inverse Agonists suggest their Potential Use in Irritable Bowel Syndrome with Constipation: A Systematic Review and Meta-Analysis.
Irritable bowel syndrome (IBS) is one of the most common functional gastrointestinal (GI) disorders characterized by pain and impaired bowel movements. Currently available drugs show limited efficacy. Cannabinoid 1 receptor (CB1) inverse agonists (CB1-RAN) cause diarrhea and may be candidates for the treatment of constipation-predominant IBS (IBS-C). We evaluated the effects of CB1-RAN in clinical trials for their potential use in IBS-C. Database search identified all clinical trials published up to May 2018 that reported rimonabant and taranabant treatment for at least one month and detailed the GI adverse events (AEs). Categorical outcomes (subgroups of AEs) were analyzed using the odds ratio (OR). Eighteen trials met the inclusion criteria. Rimonabant 20 mg produced significantly more overall AEs (OR=1.35, CI: 1.19-1.52, p<0.0001), psychiatric events (OR=1.79, CI: 1.46-2.21, p<0.001) and GI AEs (OR=2.05, CI: 1.65-2.55, p<0.001) compared to placebo. Taranabant at doses ranging from 0.5 to 8 mg produced significantly more overall AEs (OR=1.36, CI: 1.13-1.64, p<0.002), psychiatric AEs (1.82, CI: 1.54-2.16, p<0.001) and GI AEs (OR=1.75, CI: 1.29-2.37, p<0.001) compared to placebo. The approach to target CB1 in the gut for the treatment of IBS-C or chronic constipation seems a promising therapeutic option. Prospective clinical trials on the possible targeting of CB1 and the endocannabinoid system are warranted.


Enhancement of outcoupling efficiency of organic light-emitting diodes using a planarized moth-eye structure on glass substrate.
To improve optical properties of organic light-emitting diodes (OLEDs), we developed a planarized moth-eye structure by ultraviolet nanoimprint lithography (UV-NIL). The moth-eye pattern was fabricated on a glass substrate by UV-NIL using AMONIL polymer resin. It contains nanoscaled, cone-shaped, periodic pattern arrays with a 250 nm diameter and a 550 nm pitch. The glass substrate with the moth-eye structure exhibited a 1%-2% improvement in transmittance of visible wavelengths, but it requires a planarized layer with high-refractive-index materials. Photosensitive titanium precursor resin annealed by UV was used to improve the current efficiency by up to 26% versus conventional OLEDs.


New fluorescence techniques for high-throughput drug discovery.
The rapid increase of compound libraries as well as new targets emerging from the Human Genome Project require constant progress in pharmaceutical research. An important tool is High-Throughput Screening (HTS), which has evolved as an indispensable instrument in the pre-clinical target-to-IND (Investigational New Drug) discovery process. HTS requires machinery, which is able to test more than 100,000 potential drug candidates per day with respect to a specific biological activity. This calls for certain experimental demands especially with respect to sensitivity, speed, and statistical accuracy, which are fulfilled by using fluorescence technology instrumentation. In particular the recently developed family of fluorescence techniques, FIDA (Fluorescence Intensity Distribution Analysis), which is based on confocal single-molecule detection, has opened up a new field of HTS applications. This report describes the application of these new techniques as well as of common fluorescence techniques--such as confocal fluorescence lifetime and anisotropy--to HTS. It gives experimental examples and presents advantages and disadvantages of each method. In addition the most common artifacts (auto-fluorescence or quenching by the drug candidates) emerging from the fluorescence detection techniques are highlighted and correction methods for confocal fluorescence read-outs are presented, which are able to circumvent this deficiency.