Low-fat dietary pattern and risk of benign proliferative breast disease: a randomized, controlled dietary modification trial.
Modifiable factors, including diet, might alter breast cancer risk. We used the Women's Health Initiative Dietary Modification trial to test the effect of the intervention on risk of benign proliferative breast disease, a condition associated with increased risk of, and considered to be on the pathway to, invasive breast cancer. The Women's Health Initiative Dietary Modification trial was a randomized, controlled, primary prevention trial conducted in 40 U.S. clinical centers from 1993 to 2005. A total of 48,835 postmenopausal women, ages 50 to 79 years, without prior breast cancer, were enrolled. Participants were randomly assigned to the dietary modification intervention group or to the comparison group. The intervention was designed to reduce total dietary fat intake to 20% of total energy intake, and to increase fruit and vegetable intake to > or =5 servings/d and intake of grain products to > or =6 servings/d, but resulted in smaller, albeit significant, changes in practice. Participants had biennial mammograms and regular clinical breast exams. We identified women who reported breast biopsies free of cancer, obtained the histologic sections, and subjected them to standardized central review. During follow-up (average, 7.7 years), 570 incident cases of benign proliferative breast disease were ascertained in the intervention group and 793 in the comparison group. The hazard ratio for the association between dietary modification and benign proliferative breast disease was 1.09 (95% confidence interval, 0.98-1.23). Risk varied by levels of baseline total vitamin D intake but it varied little by levels of other baseline variables. These results suggest that a modest reduction in fat intake and increase in fruit, vegetable, and grain intake do not alter the risk of benign proliferative breast disease.


Aging and changes in complexity in the neurobehavioral system.
The issue of complexity is more and more present in numerous domains of biological research, including aging research. In the present paper, based on a selective review of literature, we propose both a conceptual and a methodological framework to address age-related changes in functional complexity of the neurobehavioral system, presumably resulting from modifications of the coupling between cognitive and sensorimotor processes. In particular, after reviewing pioneering and more recent studies on aging and complexity in the neuromusculoskeletal system, we explore the possibility that an age-induced increase in the coupling between cognitive and sensorimotor domains could be captured by a stronger covariation of high-order variables, common to both cognitive and sensorimotor functioning. Our main assumption is that these variables could behave as neurobehavioral markers of aging in the neuromusculoskeletal system. The present approach markedly differs from other traditional approaches, which focused on process-specific variable correlates of chronological age, domain-by-domain, and task-by-task. It provides a coherent conceptual framework, a terminology, and a method for studying age-related coupling of cognitive and sensorimotor processes with the use of complexity and nonlinear dynamical systems theories.


QTL mapping in white spruce: gene maps and genomic regions underlying adaptive traits across pedigrees, years and environments.
The genomic architecture of bud phenology and height growth remains poorly known in most forest trees. In non model species, QTL studies have shown limited application because most often QTL data could not be validated from one experiment to another. The aim of our study was to overcome this limitation by basing QTL detection on the construction of genetic maps highly-enriched in gene markers, and by assessing QTLs across pedigrees, years, and environments. Four saturated individual linkage maps representing two unrelated mapping populations of 260 and 500 clonally replicated progeny were assembled from 471 to 570 markers, including from 283 to 451 gene SNPs obtained using a multiplexed genotyping assay. Thence, a composite linkage map was assembled with 836 gene markers.For individual linkage maps, a total of 33 distinct quantitative trait loci (QTLs) were observed for bud flush, 52 for bud set, and 52 for height growth. For the composite map, the corresponding numbers of QTL clusters were 11, 13, and 10. About 20% of QTLs were replicated between the two mapping populations and nearly 50% revealed spatial and/or temporal stability. Three to four occurrences of overlapping QTLs between characters were noted, indicating regions with potential pleiotropic effects. Moreover, some of the genes involved in the QTLs were also underlined by recent genome scans or expression profile studies.Overall, the proportion of phenotypic variance explained by each QTL ranged from 3.0 to 16.4% for bud flush, from 2.7 to 22.2% for bud set, and from 2.5 to 10.5% for height growth. Up to 70% of the total character variance could be accounted for by QTLs for bud flush or bud set, and up to 59% for height growth. This study provides a basic understanding of the genomic architecture related to bud flush, bud set, and height growth in a conifer species, and a useful indicator to compare with Angiosperms. It will serve as a basic reference to functional and association genetic studies of adaptation and growth in Picea taxa. The putative QTNs identified will be tested for associations in natural populations, with potential applications in molecular breeding and gene conservation programs. QTLs mapping consistently across years and environments could also be the most important targets for breeding, because they represent genomic regions that may be least affected by G × E interactions.


A systematic review on the effectiveness of pharmacological interventions for chronic non-specific low-back pain.
The objective of this review was to determine the effectiveness of pharmacological interventions [i.e., non-steroid anti-inflammatory drugs (NSAIDs), muscle relaxants, antidepressants, and opioids] for non-specific chronic low-back pain (LBP). Existing Cochrane reviews for the four interventions were screened for studies fulfilling the inclusion criteria. Then, the literature searches were updated. Only randomized controlled trials on adults (≥18 years) with chronic (≥12 weeks) non-specific LBP and evaluation of at least one of the main clinically relevant outcome measures (pain, functional status, perceived recovery, or return to work) were included. The GRADE approach was used to determine the quality of evidence. A total of 17 randomized controlled trials was included: NSAIDs (n = 4), antidepressants (n = 5), and opioids (n = 8). No studies were found for muscle relaxants; 14 studies had a low risk of bias. The studies only reported effects on the short term (<3 months). The overall quality of the evidence was low. NSAIDs and opioids seem to lead to a somewhat higher relief in pain on the short term, as compared to placebo, in patients with non-specific chronic low back pain; opioids seem to have a small effect in improving function for a selection of patients who responded with an exacerbation of their symptoms after stopping their medication. However, both types of medication show more adverse effects than placebo. There seems to be no difference in effect between antidepressants and placebo in patients with non-specific chronic LBP.


Twin Conception in Sheep Leads to Impaired Insulin Sensitivity and Sexually Dimorphic Adipose Tissue and Skeletal Muscle Phenotypes in Adulthood.
Twins are often born small and early and have increased risk of obesity and diabetes later in life. Twin conception in sheep, regardless of whether the pregnancy continues as twins or is reduced to singleton in early gestation, alters offspring growth trajectory and body composition in young adulthood. We hypothesized that twin conception would result in insulin resistance in adulthood, with insulin-resistant adipose tissue and skeletal muscle phenotypes. At 3 years of age, body weight was not different among singletons, twins, and reductions; females weighed less than males. Singletons were leaner than reductions, with twins intermediate. Twins and reductions had decreased insulin sensitivity compared with singletons (singletons: mean [standard error of the mean]: 4.75 [0.4], twins: 3.34 [0.3], reductions: 3.67 [0.2] mg·I μU-1·kg-1·min-1, P < .01). There were no group differences in adipocyte size, adipose tissue, or circulating tumor necrosis factor α, monocyte chemoattractant protein 1, or interleukin 6 concentrations. In males, omental and subcutaneous adipose SLC2A4 was 1.5- to 2.0-fold greater in twins and reductions than in singletons ( P < .01) and SLC2A1 was greater in reductions than in singletons. Skeletal muscle IRS-1 was decreased in male twins but increased in female twins, compared to singletons ( P ≤ .01), with no effect on reductions in either sex. Skeletal muscle SLC2A4 was decreased in female twins and reductions but elevated in male twins and reductions compared to singletons ( P ≤ .01). We conclude that adult twin insulin resistance is not due to adipose tissue phenotype, but potentially phenotypic effects in skeletal muscle, and obesity is a result of twin conception per se with its origins in early gestation.


A combined digital and stereophotogrammetric technique for rehabilitation with immediate loading of complete-arch, implant-supported prostheses: A randomized controlled pilot clinical trial.
Traditional impressions for complete-arch restorations are complex and time-consuming, and they can be uncomfortable for the patient. New digital techniques such as stereophotogrammetry may mitigate this. The purpose of this randomized controlled pilot clinical trial was to compare the patient and dentist satisfaction and work times of traditional impressions (control group) and digital impressions with stereophotogrammetry in complete-arch, implant-supported prostheses. Success rates, implant survival, marginal bone loss around the dental implants, and prosthesis survival were also analyzed. This randomized controlled pilot clinical trial included 18 participants who received 131 dental implants. Implant impressions in the experimental group were made with stereophotogrammetry (8 participants with 66 implants), while traditional impressions were made in the control group (10 participants with 65 implants). Working times were measured in minutes starting from removal of the healing abutments to their replacement after the impression. Patient and dentist satisfaction was analyzed using a questionnaire with a visual analog scale, and implant success was assessed using the Buser success criteria. Prosthesis survival was defined as the presence of the prosthesis in the mouth, without screw loosening or fracture. The work times were 15.6 (experimental group) and 20.5 minutes (control group) (P<.001). The patient satisfaction scores were 8.8 in the experimental and 7.9 in the control group (P=.02). The dentist satisfaction scores were 9.1 in the experimental group and 8.5 in the control group (P=.03). The implant success rate was 100% in both groups. Marginal bone loss was 0.6 ±0.5 mm (experimental group) and 0.6 ±0.2 mm (control group) (P=.72). Digital impressions using stereophotogrammetry may be an alternative to traditional impressions. Patient and dentist satisfaction improved, and the work time was reduced in the experimental group. No statistically significant differences were found in terms of the implant success rate, implant survival, marginal bone loss, or prosthesis survival between the 2 groups.


Issues in paleopathology and possible strategies for dealing with them.
Human paleopathology has the potential of enriching what we can reconstruct about the lives of our recent and ancient ancestors. The development of remarkably sensitive analytical methods in chemistry and physics provides information that is still in the early stages of being used in research about human paleopathology. Research on mummy and skeletal tissues, both gross and histological, has vastly improved from the early studies in paleopathology. There are, however, nagging issues that have been identified as scientists attempt to conduct new research and interpret the research being done. These include the need for a greater emphasis on rigor in diagnosis of skeletal and soft tissue abnormalities; the establishment of additional research centers that recover and curate large, carefully documented archaeological samples of human remains; better integration between the medical fields of orthopaedic pathology and skeletal radiology with paleopathology; and improved design in the research conducted.


Using accelerometer feedback to identify walking destinations, activity overestimates, and stealth exercise in obese and nonobese individuals.
Accelerometer output feedback might enable assessment of recall biases for moderate bouts by obese and nonobese individuals; accelerometry might also help residents recall destinations for moderate-intensity walking bouts. Adult residents' 1-week accelerometer-measured physical activity and obesity status were measured before and after a new rail stop opened (n = 51 Time 1; n = 47 Time 2). Participants recalled the week's walking bouts, described them as brisk (moderate) or not, and reported a rail stop destination or not. At the end of the week, we provided accelerometry output to residents as a prompt. Recall of activity intensity was accurate for about 60% of bouts. Nonobese participants had more moderate bouts and more "stealth exercise" --moderate bouts recalled as not brisk--than did obese individuals. Obese participants had more overestimates--recalling light bouts as brisk walks--than did nonobese individuals. Compared with unprompted recall, accelerometry-prompted recalls allowed residents to describe where significantly more moderate bouts of activity occurred. Coupling accelerometry feedback with self-report improves research by measuring the duration, intensity, and destination of walking bouts. Recall errors and different patterns of errors by obese and nonobese individuals underscore the importance of validation by accelerometry.


Nintedanib inhibits fibroblast activation and ameliorates fibrosis in preclinical models of systemic sclerosis.
Nintedanib is a tyrosine kinase inhibitor that has recently been shown to slow disease progression in idiopathic pulmonary fibrosis in two replicate phase III clinical trials. The aim of this study was to analyse the antifibrotic effects of nintedanib in preclinical models of systemic sclerosis (SSc) and to provide a scientific background for clinical trials in SSc. The effects of nintedanib on migration, proliferation, myofibroblast differentiation and release of extracellular matrix of dermal fibroblasts were analysed by microtitre tetrazolium and scratch assays, stress fibre staining, qPCR and SirCol assays. The antifibrotic effects of nintedanib were evaluated in bleomycin-induced skin fibrosis, in a murine sclerodermatous chronic graft-versus-host disease model and in tight-skin-1 mice. Nintedanib dose-dependently reduced platelet-derived growth factor-induced and transforming growth factor-β-induced proliferation and migration as well as myofibroblast differentiation and collagen release of dermal fibroblasts from patients with and healthy individuals. Nintedanib also inhibited the endogenous activation of SSc fibroblasts. Nintedanib prevented bleomycin-induced skin fibrosis in a dose-dependent manner and was also effective in the treatment of established fibrosis. Moreover, treatment with nintedanib ameliorated fibrosis in the chronic graft-versus-host disease model and in tight-skin-1 mice in well-tolerated doses. We demonstrate that nintedanib effectively inhibits the endogenous as well as cytokine-induced activation of SSc fibroblasts and exerts potent antifibrotic effects in different complementary mouse models of SSc. These data have direct translational implications for clinical trials with nintedanib in SSc.


Formation of tissue factor-factor VIIa-factor Xa complex prevents apoptosis in human breast cancer cells.
Tissue factor (TF) is a transmembrane glycoprotein that initiates blood coagulation when complexed with factor VIIa (FVIIa). TF is constitutively expressed in a variety of tumor cells and has been shown to play a role in cellular signaling and tumor progression. In this study, we investigated the effect of TF-FVIIa mediated signaling on apoptosis in human breast cancer cells. Apoptosis was induced by prolonged serum starvation and studied using the Adr-MCF-7 cell line, which has high endogenous TF expression. Treatment of the cells with the combination of FVIIa (10 nM) and FX (150 nM), reduced apoptosis by nearly 50% compared with untreated, control cells using an ELISA that detects histone-DNA fragments. In contrast, FVIIa (10 nM) alone did not significantly prevent apoptosis. Pretreatment of the Adr-MCF-7 cells with hirudin, a specific thrombin inhibitor, did not inhibit the anti-apoptotic effect of the combination of FVIIa and FX, whereas this effect could be abrogated by inhibition of phosphorylation of either p44/42 mitogen-activated protein kinase (MAPK) or protein kinase B (PKB/Akt). In addition, treatment of the Adr-MCF-7 cells with the combination of FVIIa and FX led to a 30-50% increase in the level of the anti-apoptotic protein, survivin, compared with untreated cells using Western blot analysis. These results indicate that formation of TF-FVIIa-FXa complex prevents apoptosis in breast cancer cells by a thrombin-independent pathway. Moreover, the anti-apoptotic effect of this signaling pathway involves phosphorylation of both p44/42 MAPK and PKB/Akt and might be mediated in part by an increase in cell survivin levels.


Information theory-based automatic multimodal transfer function design.
In this paper, we present a new framework for multimodal volume visualization that combines several information-theoretic strategies to define both colors and opacities of the multimodal transfer function. To the best of our knowledge, this is the first fully automatic scheme to visualize multimodal data. To define the fused color, we set an information channel between two registered input datasets, and afterward, we compute the informativeness associated with the respective intensity bins. This informativeness is used to weight the color contribution from both initial 1-D transfer functions. To obtain the opacity, we apply an optimization process that minimizes the informational divergence between the visibility distribution captured by a set of viewpoints and a target distribution proposed by the user. This distribution is defined either from the dataset features, from manually set importances, or from both. Other problems related to the multimodal visualization, such as the computation of the fused gradient and the histogram binning, have also been solved using new information-theoretic strategies. The quality and performance of our approach are evaluated on different datasets.


Curcumin attenuates aluminium-induced functional neurotoxicity in rats.
Curcumin is a polyphenol extracted from the rhizome of Curcuma longa and well known as a multi-functional drug with antioxidative, anti-cancerous and anti-inflammatory activities. Curcumin's antiaging and neuroprotective potential is widely reported. In the present study, effect of curcumin treatment dose 30 mg kg(-1) day(-1) was investigated against aluminium neurotoxicity in young and old animals. Direct and indirect intakes of aluminium have been reported to be involved in the etiology of several neurodegenerative disorders like Alzheimer's and Parkinson's diseases. Long term Al was administered through drinking water at a dose of 50 mg/kg/day for 6 months in both young (4 months) and old (18 months) male Wistar rats. Result obtained demonstrates that curcumin treatment attenuates the Al-induced alterations at biochemical, behavioral and ultrastructural levels which was well reflected in the electrophysiological recordings. Our results indicate that curcumin's ability to bind redox active metals and cross the blood-brain barrier could be playing crucial role in preventing against Al-induced neurotoxicity.


Assessment of Operative Autonomy and Readiness for Independent Practice Among Pediatric Surgery Fellows.
The purpose of this study was to evaluate the implementation of the System for Improving and Measuring Procedural Learning (SIMPL) "app" in a pediatric surgery training program. SIMPL is a novel workplace-based operative performance assessment platform which allows case by case assessment of trainees using three scales (autonomy, performance, and case complexity) and takes seconds to complete. A pediatric surgery-specific procedural taxonomy for SIMPL was developed from the ACGME procedural library. SIMPL was piloted and implemented in a single pediatric surgery training program. A descriptive analysis of the operative assessments was undertaken. In the year following SIMPL implementation, 565 (39% of cases) assessments were completed by 21 faculty for 3 fellows for 148 unique procedures within a median of 8.5 h. Recorded feedback accompanied 61% of assessments. 2nd year fellows were more likely to be deemed autonomous (81% ratings) and practice ready (84%) vs. 1st year fellows (50%; p < 0.001 both), with improvements over time. Within pediatric surgery, this is the first implementation of a workplace-based operative performance assessment "app". With its ease of use, SIMPL drastically increased the volume of operative evaluations and diversified the mix of cases evaluated while SIMPL ratings demonstrated expected and graduated performance improvements over time. Education. N/A.


Revised Starling equation and the glycocalyx model of transvascular fluid exchange: an improved paradigm for prescribing intravenous fluid therapy.
I.V. fluid therapy does not result in the extracellular volume distribution expected from Starling's original model of semi-permeable capillaries subject to hydrostatic and oncotic pressure gradients within the extracellular fluid. Fluid therapy to support the circulation relies on applying a physiological paradigm that better explains clinical and research observations. The revised Starling equation based on recent research considers the contributions of the endothelial glycocalyx layer (EGL), the endothelial basement membrane, and the extracellular matrix. The characteristics of capillaries in various tissues are reviewed and some clinical corollaries considered. The oncotic pressure difference across the EGL opposes, but does not reverse, the filtration rate (the 'no absorption' rule) and is an important feature of the revised paradigm and highlights the limitations of attempting to prevent or treat oedema by transfusing colloids. Filtered fluid returns to the circulation as lymph. The EGL excludes larger molecules and occupies a substantial volume of the intravascular space and therefore requires a new interpretation of dilution studies of blood volume and the speculation that protection or restoration of the EGL might be an important therapeutic goal. An explanation for the phenomenon of context sensitivity of fluid volume kinetics is offered, and the proposal that crystalloid resuscitation from low capillary pressures is rational. Any potential advantage of plasma or plasma substitutes over crystalloids for volume expansion only manifests itself at higher capillary pressures.


Curcumin Reactivates Silenced Tumor Suppressor Gene RARβ by Reducing DNA Methylation.
Reactivation of tumor suppressor genes by nontoxic bioactive food component represents a promising strategy for cancer chemoprevention. Retinoic acid receptor β (RARβ), one member of the RAR receptor family, is considered as a tumor suppressor. Reduced expression of RARβ has been reported in lung cancer and other solid tumors. DNA hypermethylation of the promoter region of RARβ is a major mechanism for its silencing in tumors. Recently, curcumin has been considered as a potential DNA methyltransferase inhibitor. Herein, we demonstrated that curcumin significantly elevate RARβ expression at the mRNA and protein levels in tested cancer cells. Additionally, curcumin decreased RARβ promoter methylation in lung cancer A549 and H460 cells. Mechanistic study demonstrated that curcumin was able to downregulate the mRNA levels of DNMT3b. In a lung cancer xenograft node mice model, curcumin exhibited protective effect against weight loss because of tumor burden. Tumor growth was strongly repressed by curcumin treatment. As the results from in vitro, RARβ mRNA were increased and DNMT3b mRNA were decreased by curcumin treatment compared with the mice in control group. Altogether, this study reveals a novel molecular mechanism of curcumin as a chemo-preventive agent for lung cancer through reactivation of RARβ.


Meta-analysis on the effects of the physical environment, animal traits, feeder and feed characteristics on the feeding behaviour and performance of growing-finishing pigs.
A meta-analysis, using information from 45 experiments on growing-finishing pigs published in 39 manuscripts, was carried out to determine the simultaneous effects of the physical environment (space allowance, group size, flooring conditions, temperature, presence of enrichment), pig traits (initial body weight (BW) for each studied time interval, sex, genetics), feeder characteristics (water provision within the feeder, feeder design (individual/collective), feeder places/pig, presence of feeder protection) and feed characteristics (feed allowance (ad libitum/restricted), net energy content, crude protein (CP) content), as well as their potential interactions, on the feeding behaviour and performance of growing-finishing pigs. The detrimental effect of low temperature on performance was particularly evident for restricted-fed pigs (P < 0.05). At reduced feeder space allowance, a reduction in the percentage of time spent eating was predicted when increasing initial BW, whereas the opposite was predicted for larger feeder space allowances (P < 0.001). The reduction in visit duration to the feeder in higher BW groups became gradually more important with increasing feeder space allowance (P < 0.01), whereas the increase in the ingestion rate and average daily feed intake (ADFI) with increasing initial BW became smaller with increasing feeder space (P < 0.05). The model predicted a reduction in feed conversion ratio (FCR) with increasing group size (P < 0.05) and floor space allowance (P < 0.01) and on solid floors with or without bedding (P < 0.05). In comparison with other feeders, wet/dry feeders were associated with more frequent but shorter feeder visits (P < 0.05), higher ingestion rates (P < 0.001) and higher ADFI (P < 0.10). The use of protection within individual feeders increased the time spent feeding (P < 0.001), reduced the number of visits per day (P < 0.01), the ingestion rate (P < 0.001) and FCR (P < 0.01) in comparison with other feeder types. Sex modulated the effect of the number of feeder places/pig on FCR (P < 0.05), with a gradual reduction of FCR in entire males and females when increasing feeder space allowance. Genetics tended to modulate the effect of diets' CP content on FCR (P < 0.10). Overall, these results may contribute to the improvement of the welfare and performance of growing-finishing pigs by a better knowledge of the influence of the rearing environment and may help optimize the feeding strategies in current production systems.


Mice with a deletion of the major central myelin protein exhibit hypersensitivity to noxious thermal stimuli: involvement of central sensitization.
Null mutations in the gene encoding the major myelin protein of the central nervous system, proteolipid protein 1 (PLP1), cause an X-linked form of spastic paraplegia (SPG2) associated with axonal degeneration. While motor symptoms are the best known manifestations of this condition, its somatosensory disturbances have been described but poorly characterized. We carried out a longitudinal study in an animal model of SPG2 - mice carrying a deletion of the Plp1 gene (Plp-null mice). Plp-null mice exhibited severe early-onset thermal hyperalgesia, in the absence of thermal allodynia. We first performed an electrophysiological testing which showed an early decrease in peripheral and spinal conduction velocities in Plp null mice. Such as the abnormal sensitive behaviors, this slowing of nerve conduction was observed before the development of myelin abnormalities at the spinal level, from 3months of age, and without major morphological defects in the sciatic nerve. To understand the link between a decrease in nerve velocity and an increased response to thermal stimuli before the appearance of myelin abnormalities, we focused our attention on the dorsal horn of the spinal cord, the site of integration of somatosensory information. Immunohistochemical studies revealed an early-onset activation of astrocytes and microglia that worsened with age, associated later in age with perturbation of the expression of the sensory neuropeptides calcitonin-gene-related peptide and galanin. Taken together, these results represent complementary data supporting the hypothesis that Plp-null mice suffer from ganglionopathy associated with late onset central demyelination but with few peripheral nerve alterations, induced by the glial-cell-mediated sensitization of the spinal cord. The mechanism suggested here could underlie pain experiments in other leukodystrophies as well as in other non-genetic demyelinating diseases such as multiple sclerosis.


Incisive structure-spectroscopic correlation in oligothiophenes functionalized with (+/-) inductive/mesomeric fluorine groups: joint Raman and DFT study.
This paper presents a combined experimental Raman and density functional theory (DFT) study of a series of oligothiophenes with variable pi-core lengths and substituted at the alpha,omega- and beta,beta'-terminal positions with perfluorohexyl, perfluorohexylcarbonyl, perfluoroarene, and cyano groups. The study covers the neutral and the electrochemically oxidized samples. The spectra have been assigned with the help of B3LYP/6-31G** calculations and interpreted by the predictions of effective conjugation coordinate (ECC) theory. Direct relationships between the bond length alternation (BLA) parameters and the Raman wavenumbers of the two most important bands of the spectra have been outlined showing the collective character of their electronic and vibrational properties. It is found that the topology of the thienyl electronic structure is not uniform along the conjugated core with the external rings more aromatic, whereas for the innermost part the pi-electron delocalization is greater and extends with the increment of chain length. Perfluorohexyl substitution finely tunes the electronic properties via negative inductive effects. The beta,beta' isomers exhibit larger conformational distortion, which strongly modifies the mean conjugation length. Oxidation provokes electronic structure quinoidization spreading over the whole system but more marked in the central part of the pi-core. The Raman features associated to quinoidization in the oxidized species have been interpreted in relation to the oligomer core length and the substitution regiochemistry.


Obesity is associated with greater inflammation and monocyte activation among HIV-infected adults receiving antiretroviral therapy.
Among virally suppressed HIV-infected persons, we examined the relationship between obesity and alterations in key clinical markers of immune activation and inflammation. These markers have also been associated with excess HIV-related cardiovascular disease and mortality. We evaluated data from virally suppressed participants in the Study to Understand the Natural History of HIV/AIDS in the Era of Effective Therapy, including inflammatory biomarkers (interleukin-6 and highly sensitive C-reactive protein), monocyte biomarkers [soluble CD163 (sCD163), sCD14], and monocyte immunophenotypes. We assessed associations with these immunologic measures and obesity, via logistic regression preadjustment and postadjustment for demographic and clinical factors, homeostatic model assessment of insulin resistance, and leptin levels. Among 452 evaluable participants, median (interquartile range) age was 41 (36-48) years, CD4 cell count was 475 (308-697) cells/μl, and 21% were obese (BMI ≥ 30 kg/m). In univariable models, obesity, smoking, and lower CD4 cell count were associated with higher measures of inflammation and monocyte activation. After adjustment, obesity remained independently associated with elevated levels (highest vs. lower two tertiles) of interleukin-6 [odds ratio (OR) 1.96; P = 0.02], highly sensitive C-reactive protein (OR 2.79; P < 0.001) and sCD163 (OR 1.94; P = 0.02), and elevated frequency of CD14CD16 (OR 1.77; P = 0.03) and CD14dimCD16 (OR 1.97; P = 0.01). Adjusting for homeostatic model assessment of insulin resistance and leptin modestly affected associations for obesity with inflammation and monocyte activation. Obesity was prevalent and independently associated with greater monocyte activation and systemic inflammation. Research is needed to determine how adipose tissue excess is functionally related to persistent immunologic abnormalities among HIV-infected persons with viral suppression.


Dendritic pathology of granule cells in Alzheimer's disease is unrelated to neuritic plaques.
Neuritic plaques are the histologic hallmark of Alzheimer's disease (AD); however, the extent to which they are injurious to neurons is unclear. In order to investigate this problem, we intracellularly filled human dentate granule cells with Lucifer yellow in a lightly fixed slice preparation and studied the relationships between their dendrites and neuritic plaques. After counterstaining for plaques and drawing the filled granule cell dendrites, we found that there were significant differences in the morphology of dendrites in control and AD cases; granule cell dendrites from Alzheimer's cases were generally shorter, branched less profusely, and had fewer spines than those from age matched controls. Surprisingly, when dendrites traveled into plaques, they still bore spines and their morphology was distinct from that of the amyloid-stained dystrophic neurites surrounding them. Furthermore, within AD cases we found no significant differences between dendrites that were located directly beneath or passing through plaques and those that were located in plaque-free regions. We conclude that granule cell dendrites are not an integral component of plaques within their dendritic fields and that neuritic plaques have no direct effect on granule cells in the dentate gyrus.


Prostate cancer treatment is enhanced by genistein in vitro and in vivo in a syngeneic orthotopic tumor model.
Pretreatment with genistein, a bioactive component of soy isoflavones, potentiated cell killing induced by radiation in human PC-3 prostate cancer cells in vitro. Using an orthotopic xenograft in nude mice, we demonstrated that genistein combined with prostate tumor irradiation caused greater inhibition of primary tumor growth and increased control of spontaneous metastasis to para-aortic lymph nodes, increasing mouse survival. Paradoxically, treatment with genistein alone increased metastasis to lymph nodes. This observation is of concern in relation to soy-based clinical trials for cancer patients. To address whether this observation is because nude mice have an impaired immune system, these studies were repeated in orthotopic RM-9 prostate tumors in syngeneic C57BL/6 mice. The combination of genistein with radiation in this model also caused a greater inhibition of primary tumor growth and spontaneous metastasis to regional para-aortic lymph nodes, whereas treatment with genistein alone showed a trend to increased lymph node metastasis. Data from the syngeneic and xenograft models are comparable and indicate that the combination of genistein with radiotherapy is more effective and safer for prostate cancer treatment than genistein alone, which promotes metastatic spread to regional lymph nodes.


Lack of ADAM2, CALR3 and SAGE1 Cancer/Testis Antigen Expression in Lung and Breast Cancer.
Immunotherapy is emerging as a supplement to conventional cancer treatment, and identifying antigen targets for specific types of cancer is critical to optimizing therapeutic efficacy. Cancer/testis antigens are highly promising targets for immunotherapy due to their cancer-specific expression and antigenic properties, but the expression patterns of most of the more than 200 identified cancer/testis antigens in various cancers remain largely uncharacterized. In this study, we investigated the expression of the cancer/testis antigens ADAM2, CALR3 and SAGE1 in lung and breast cancer, the two most frequent human cancers, with the purpose of providing novel therapeutic targets for these diseases. We used a set of previously uncharacterized antibodies against the cancer/testis antigens ADAM2, CALR3 and SAGE1 to investigate their expression in a large panel of normal tissues as well as breast and lung cancers. Staining for the well-characterized MAGE-A proteins was included for comparison. Immunohistochemical staining confirmed previous mRNA analysis demonstrating that ADAM2, CALR3 and SAGE1 proteins are confined to testis in normal individuals. Negative tissues included plancenta, which express many other CT antigens, such as MAGE-A proteins. Surprisingly, we detected no ADAM2, CALR3 and SAGE1 in the 67 lung cancers (mainly non-small lung cancer) and 189 breast cancers, while MAGE-A proteins were present in 15% and 7-16% of these tumor types, respectively. Treatment with DNA methyltransferase inhibitors has been proposed as an attractive strategy to increase the expression of cancer/testis antigens in tumors before immunotargeting; however, neither ADAM2, CALR3 nor SAGE1 could be significantly induced in lung and breast cancer cell lines using this strategy. Our results suggest that ADAM2, CALR3 and SAGE1 cancer/testis antigens are not promising targets for immunotherapy of breast and lung cancer.


Analysis of the genetic basis of disease in the context of worldwide human relationships and migration.
Genetic diversity across different human populations can enhance understanding of the genetic basis of disease. We calculated the genetic risk of 102 diseases in 1,043 unrelated individuals across 51 populations of the Human Genome Diversity Panel. We found that genetic risk for type 2 diabetes and pancreatic cancer decreased as humans migrated toward East Asia. In addition, biliary liver cirrhosis, alopecia areata, bladder cancer, inflammatory bowel disease, membranous nephropathy, systemic lupus erythematosus, systemic sclerosis, ulcerative colitis, and vitiligo have undergone genetic risk differentiation. This analysis represents a large-scale attempt to characterize genetic risk differentiation in the context of migration. We anticipate that our findings will enable detailed analysis pertaining to the driving forces behind genetic risk differentiation.


Long-Term Maintenance of Therapeutic Gains Associated With Cognitive-Behavioral Therapy for Insomnia Delivered Alone or Combined With Zolpidem.
To document the long-term sleep outcomes at 12 and 24 months after patients with chronic insomnia were treated with cognitive-behavioral therapy (CBT), either singly or combined with zolpidem medication. Participants were 160 adults with chronic insomnia. They were first randomized for a six-week acute treatment phase involving CBT alone or CBT combined with nightly zolpidem, and randomized for a six-month extended treatment phase involving CBT, no additional treatment, CBT combined with zolpidem as needed, or CBT with zolpidem tapered. This paper reports results of the 12- and 24-month follow-ups on the main outcome measures derived from the Insomnia Severity Index and sleep diaries. Clinical improvements achieved 6 months following the end of treatment were well-maintained in all four conditions, with insomnia remission rates ranging from 48% to 74% at the 12-month follow-up, and from 44% to 63% at the 24-month follow-up. Participants receiving CBT with zolpidem taper in the extended treatment phase had significantly better results than those receiving CBT with continued zolpidem as needed. The magnitude of improvements on sleep diary parameters was similar between conditions, with a slight advantage for the CBT with zolpidem taper condition. The addition of extended CBT did not alter the long-term outcome over improvements obtained during the initial 6-week CBT. The results suggest that CBT for insomnia, when delivered alone or in combination with medication, produce durable sleep improvements up to two years after completion of treatment. These long-term results indicate that even if a combined CBT plus medication approach provide an added benefit immediately after treatment, extending CBT while tapering medication produce better sustained improvements compared to continued use of medication as needed.


Bone marrow oedema assessment by magnetic resonance imaging in rheumatoid arthritis wrist and metacarpophalangeal joints: the importance of field strength, coil type and image resolution.
The aim of this study was to examine the influence of different MRI unit field strengths, coil types and image resolutions on the OMERACT RA MRI scoring system (RAMRIS) of bone marrow oedema (BME) and image quality. Forty-one patients and 12 healthy controls participated in this cross-sectional study. Coronal short tau inversion recovery (STIR) and T1-weighted sequences were obtained at 0.23, 0.6, 1.5 and 3T using flex coils (Flex). Additional STIR sequences were obtained with phased array extremity coils (Extr) (at 0.6 and 1.5T) and higher resolution (at 1.5T). In otal, 338 STIR image sets were anonymized and scored according to RAMRIS and parameters of image quality were measured. The BME sum scores were similar overall when comparing the different MRI units, coil types and voxel sizes, yet significantly higher at the higher resolution of 1.5T Extr compared with 0.23T Flex (P = 0.004), 0.6T Flex (P = 0.03), 1.5T Flex (P = 0.05) and 3T Flex (P = 0.001). Mean differences were relatively minor (0-3.5). Intrareader reliability of BME scores was high [intraclass correlation coefficient ≥ 0.90 for all except 0.23T (0.81) and percentage exact agreement 81-88%]. The smallest detectable difference was better at 0.6, 1.5 and 3T (9-29% of maximum value) than at 0.23T (40%). Image quality was lowest at 0.23T. No major, consistent differences were found between BME scores using STIR sequences obtained at different field strengths, coil types and image resolutions, suggesting that these are equally suited for assessment of BME in RA. However, parameters of image quality and intrareader reliability (favouring 0.6, 1.5 and 3T) should be considered when selecting the MRI acquisition strategy.


Adapalene pretreatment increases follicular penetration of clindamycin: in vitro and in vivo studies.
Topical retinoids normalize desquamation, reduce comedogenesis and may enhance the penetration of other topicals providing more effective treatment of acne. We evaluated the effect of adapalene on skin penetration of clindamycin phosphate when it is applied concomitantly or after various time durations following adapalene application. The in vitro studies were carried out using excised rat skin, whereas the in vivo studies were conducted on healthy human volunteers. Radioactive clindamycin phosphate (1%) gel was applied to rat skin sections and to the hands of human volunteers concomitantly and after the pretreatment of the skin for 3, 5 and 10 min with 10 mg of adapalene (0.1%) gel. Quantification of clindamycin phosphate was performed by liquid scintillation. In vitro skin penetration and distribution of clindamycin phosphate was affected by the pretreatment time. Significantly higher skin concentration of clindamycin phosphate (15.5%) with largest proportion in viable skin layer (9.4% of applied dose) was found when clindamycin phosphate gel was applied after the pretreatment of the skin with adapalene gel for 5 min. Further increase in pretreatment time has no additive influence on the penetration of clindamycin phosphate. In vivo results were in corroboration with the in vitro results and demonstrate significantly higher concentration of clindamycin phosphate (19%) in the skin following pretreatment with adapalene gel for 5 min. Adapalene acts as a penetration enhancer and increases the penetration of topical clindamycin phosphate. Application of clindamycin phosphate gel after the pretreatment of skin with adapalene gel for 5 min may contribute significantly to the increased efficacy of therapy.


High RBM3 expression in prostate cancer independently predicts a reduced risk of biochemical recurrence and disease progression.
High expression of the RNA-binding protein RBM3 has previously been found to be associated with good prognosis in breast cancer, ovarian cancer, malignant melanoma and colorectal cancer. The aim of this study was to examine the prognostic impact of immunohistochemical RBM3 expression in prostate cancer. Immunohistochemical RBM3 expression was examined in a tissue microarray with malignant and benign prostatic specimens from 88 patients treated with radical prostatectomy for localized disease. While rarely expressed in benign prostate gland epithelium, RBM3 was found to be up-regulated in prostate intraepithelial neoplasia and present in various fractions and intensities in invasive prostate cancer. High nuclear RBM3 expression was significantly associated with a prolonged time to biochemical recurrence (BCR) (HR 0.56, 95% CI: 0.34-0.93, p = 0.024) and clinical progression (HR 0.09, 95% CI: 0.01-0.71, p = 0.021). These associations remained significant in multivariate analysis, adjusted for preoperative PSA level in blood, pathological Gleason score and presence or absence of extracapsular extension, seminal vesicle invasion and positive surgical margin (HR 0.41, 95% CI: 0.19-0.89, p = 0.024 for BCR and HR 0.06, 95% CI: 0.01-0.50, p = 0.009 for clinical progression). Our results demonstrate that high nuclear expression of RBM3 in prostate cancer is associated with a prolonged time to disease progression and, thus, a potential biomarker of favourable prognosis. The value of RBM3 for prognostication, treatment stratification and follow-up of prostate cancer patients should be further validated in larger studies.


High-resolution, dual-depth spectral-domain optical coherence tomography with interlaced detection for whole-eye imaging.
Dual-depth spectral-domain optical coherence tomography (SD-OCT) enables high-resolution in vivo whole-eye imaging. Two orthogonally polarized beams from a source are focused simultaneously on two axial positions of the anterior segment and the retina. For the detector arm, a 1×2 ultrafast optical switch sequentially delivers two spectral interference signals to a single spectrometer, which extends the in-air axial depth range up to 9.44 mm. An off-pivot complex conjugate removal technique doubles the depth range for all anterior segment imaging. The graphics-processing-unit-based parallel signal processing algorithm supports fast two- and three-dimensional image displays. The obtained high-resolution anterior and retinal images are measured biometrically. The dual-depth SD-OCT system has an axial resolution of ∼6.4 μm in air, and the sensitivity is 91.79 dB at 150 μm from the zero-delay line.


Diversity-generating retroelements.
Parasite adaptation to dynamic host characteristics is a recurrent theme in biology. Diversity-generating retroelements (DGRs) are a newly discovered family of genetic elements that function to diversify DNA sequences and the proteins they encode. The prototype DGR was identified in a temperate bacteriophage, BPP-1, on the basis of its ability to generate variability in a gene that specifies tropism for receptor molecules on host Bordetella species. Tropism switching is a template-dependent, reverse transcriptase mediated process that introduces nucleotide substitutions at defined locations within a target gene. This cassette-based mechanism is theoretically capable of generating trillions of different amino acid sequences in a distal tail fiber protein, providing a vast repertoire of potential ligand-receptor interactions. Variable residues are displayed in the context of a specialized C-type lectin fold, which has evolved a unique solution for balancing protein diversity against structural stability. Homologous DGRs have been identified in the chromosomes of diverse bacterial species. These unique genetic elements have the potential to confer powerful selective advantages to their hosts, and their ability to generate novel binding specificities and dynamic antimicrobial agents suggests numerous applications.


Novel aerobic benzene degrading microorganisms identified in three soils by stable isotope probing.
The remediation of benzene contaminated groundwater often involves biodegradation and although the mechanisms of aerobic benzene biodegradation in laboratory cultures have been well studied, less is known about the microorganisms responsible for benzene degradation in mixed culture samples or at contaminated sites. To address this knowledge gap, DNA based stable isotope probing (SIP) was utilized to identify active benzene degraders in microcosms constructed with soil from three sources (a contaminated site and two agricultural sites). For this, replicate microcosms were amended with either labeled (¹³C) or unlabeled benzene and the extracted DNA samples were ultracentrifuged, fractioned and subject to terminal restriction fragment length polymorphism (TRFLP). The dominant benzene degraders (responsible for ¹³C uptake) were determined by comparing relative abundance of TRFLP phylotypes in heavy fractions of labeled benzene (¹³C) amended samples to the controls (from unlabeled benzene amended samples). Two phylotypes (a Polaromonas sp. and an Acidobacterium) were the major benzene degraders in the microcosms constructed from the contaminated site soil, whereas one phylotype incorporated the majority of the benzene-derived ¹³C in each of the agricultural soils ("candidate" phylum TM7 and an unclassified Sphingomonadaceae).


Three-photon indirect sensitization in green upconversion luminescence of Er/Tm codoped NaY(WO(4))(2) crystal.
Upconversion spectra of Er/Tm codoped NaY(WO(4))(2) crystal excited by 974nm laser is investigated. Intensive 520nm/550nm and 800nm emissions are observed. Three-photon process at high pumping power is presented in the green upconversion. A model is proposed that the three-photon process is actualized by the energy indirect sensitization between two Er(3+) ions and one Tm(3+) ion, via excited state absorption and two cross relaxations. Two Er(3+) ions totally absorb three 974nm photons, emitting one green photon, and Tm(3+) ion plays the role of temporary energy storage in this process.


Management of Malignant Peritoneal Mesothelioma Using Cytoreductive Surgery and Perioperative Chemotherapy.
Malignant peritoneal mesothelioma is a rare disease, with approximately 800 new patients per year in the United States. Its natural history is defined by progression restricted to the peritoneal space. In the past, patients with this disease had a limited lifespan of approximately 1 year. Numerous single-institution studies as well as a systematic review have reported median survival of 3 to 5 years with a combination of cytoreductive surgery and hyperthermic perioperative chemotherapy. These markedly improved survival statistics were achieved in experienced centers with 1% mortality and 20% morbidity rates. Data have shown that knowledgeable patient selection is required to prevent patients unlikely to benefit from undergoing these interventions. The conclusion is that patients with peritoneal mesothelioma can experience long-term progression-free survival or significant palliation with cytoreductive surgery plus hyperthermic perioperative chemotherapy. This management plan should be considered the standard of care for properly selected patients with malignant peritoneal mesothelioma at experienced centers around the world.


Diffusion weighted MR imaging (DWI) and ADC values in endometrial carcinoma.
To determine the sensitivity of MR imaging utilizing diffusion weighted imaging (DWI) in detection of endometrial cancer and to compare the Apparent Diffusion Coefficient (ADC) values of malignant and normal endometrium. Cross-sectional analytical study. Radiology Department, Aga Khan University Hospital Karachi, from January 2007 to December 2009. Patients were defined as undergoing MRI for suspected endometrial malignancy, and the controls were female subjects who underwent MRI examination for indications other than endometrial malignancy. Studied variables included the signal characteristics of the endometrium and ADC values. The sensitivity of DWI for the detection of endometrial carcinoma was calculated using histopathology as the gold standard, and the ADC values of the endometrium in patients and controls were recorded. The mean ADC values were compared using two-sided t- test with significance at p < 0.05 at 95% confidence interval. Out of 52 patients, 10 had hyperintense, 40 had hypointense and 2 had isointense endometrium on T2 weighted imaging. On DWI, 42 patients had hyperintense and 10 patients had hypointense endometrium. In comparison, 40 controls had hypointense endometrium on DWI and 12 had hyperintense endometrium. The mean ADC value for abnormal endometrium was 0.730±0.215 x10-3 mm2/sec and of normal endometrium was 1.265±0.305 x10-3 mm2/sec (p < 0.001). The sensitivity for detection of endometrial carcinoma on DWI was 77.77%. False positive cases were found to be due to secretory and hyperplastic endometrium. False negative findings were found in a few cases of adenocarcinoma, endometroid carcinoma and clear cell sarcoma. In patients with suspected endometrial carcinoma, MRI examination utilizing DWI was a sensitive tool in detecting endometrial cancers with significantly lower ADC values for carcinoma in general. Physiological and histopathological variants may be responsible for a few false results.


A review of scientific research and recommendations regarding eggs.
For much of the past 40 years, the public has been warned away from eggs because of a concern over coronary heart disease risk. This concern is based on three observations: 1. eggs are a rich source of dietary cholesterol; 2. when fed experimentally, dietary cholesterol increases serum cholesterol and; 3. high serum cholesterol predicts the onset of coronary heart disease. However, data from free-living populations show that egg consumption is not associated with higher cholesterol levels. Furthermore, as a whole, the epidemiologic literature does not support the idea that egg consumption is a risk factor for coronary disease. Within the nutritional community there is a growing appreciation that health derives from an overall pattern of diet rather than from the avoidance of particular foods, and there has been a shift in the tone in recent dietary recommendations away from "avoidance" messages to ones that promote healthy eating patterns. The most recent American Heart Association guidelines no longer include a recommendation to limit egg consumption, but recommend the adoption of eating practices associated with good health. Based on the epidemiologic evidence, there is no reason to think that such a healthy eating pattern could not include eggs.


The left intraparietal sulcus modulates the selection of low salient stimuli.
Neuropsychological and functional imaging studies have suggested a general right hemisphere advantage for processing global visual information and a left hemisphere advantage for processing local information. In contrast, a recent transcranial magnetic stimulation study [Mevorach, C., Humphreys, G. W., & Shalev, L. Opposite biases in salience-based selection for the left and right posterior parietal cortex. Nature Neuroscience, 9, 740-742, 2006b] demonstrated that functional lateralization of selection in the parietal cortices on the basis of the relative salience of stimuli might provide an alternative explanation for previous results. In the present study, we applied a whole-brain analysis of the functional magnetic resonance signal when participants responded to either the local or the global levels of hierarchical figures. The task (respond to local or global) was crossed with the saliency of the target level (local salient, global salient) to provide, for the first time, a direct contrast between brain activation related to the stimulus level and that related to relative saliency. We found evidence for lateralization of salience-based selection but not for selection based on the level of processing. Activation along the left intraparietal sulcus (IPS) was found when a low saliency stimulus had to be selected irrespective of its level. A control task showed that this was not simply an effect of task difficulty. The data suggest a specific role for regions along the left IPS in salience-based selection, supporting the argument that previous reports of lateralized responses to local and global stimuli were contaminated by effects of saliency.


Information thermodynamics on causal networks.
We study nonequilibrium thermodynamics of complex information flows induced by interactions between multiple fluctuating systems. Characterizing nonequilibrium dynamics by causal networks (i.e., Bayesian networks), we obtain novel generalizations of the second law of thermodynamics and the fluctuation theorem, which include an informational quantity characterized by the topology of the causal network. Our result implies that the entropy production in a single system in the presence of multiple other systems is bounded by the information flow between these systems. We demonstrate our general result by a simple model of biochemical adaptation.


Photoperiodic modulation of adrenal gland function in the rhesus macaque: effect on 24-h plasma cortisol and dehydroepiandrosterone sulfate rhythms and adrenal gland gene expression.
In temperate zones, day length changes markedly across the year, and in many mammals these photoperiodic variations are associated with physiological adaptations. However, the influence of this environmental variable on human behavior and physiology is less clear, and the potential underlying mechanisms are unknown. To address this issue, we examined the effect of changing photoperiods on adrenal gland function in ovariectomized female rhesus macaques (Macaca mulatta), both in terms of steroid hormone output and in terms of gene expression. The animals were sequentially exposed to the following lighting regimens, which were designed to simulate photoperiods associated with winter, spring/autumn and summer respectively: 8 h light:16 h darkness (short days), 12 h light:12 h darkness and 16 h light:8 h darkness (long days). Remote 24-h serial blood sampling failed to disclose any effect of photoperiod on mean or peak plasma levels of cortisol or dehydroepiandrosterone sulfate. However, there was a marked phase-advancement of both hormonal rhythms in short days, which was reflected as a similar phase-advancement of the daily motor activity rhythm. Gene microarray analysis of the adrenal gland transcriptome revealed photoperiod-induced differences in the expression of genes associated with homeostatic functions, including: development, lipid synthesis and metabolism, and immune function. Taken together, the results indicate that in primates, both circadian adrenal physiology and gene expression are influenced by seasonal changes in day length, which may have implications for adrenal-regulated physiology and behavior.


Angular dependent anisotropic terahertz response of vertically aligned multi-walled carbon nanotube arrays with spatial dispersion.
Spatial dispersion effect of aligned carbon nanotubes (CNTs) in the terahertz (THz) region has significance for both theoretical and applied consideration due to the unique intrinsically anisotropic physical properties of CNTs. Herein, we report the angular dependent reflection of p-polarized THz wave from vertically aligned multi-walled CNT arrays in both experiment and theory. The spectra indicate that the reflection depends on the film thickness of vertically aligned CNTs, the incident angle, and the frequency. The calculation model is based on the spatial dispersion effect of aligned CNTs and performed with effective impedance method and the Maxwell-Garnett approximation. The results fit well with the experiment when the thickness of CNT film is thin, which reveals a coherent superposition mechanism of the CNT surface reflection and CNTs/Si interface reflection. For thick CNT films, the CNTs/Si interface response determines the reflection at small incident angles, while the CNTs surface effect dominates at large incident angles. This work investigates the spatial dispersion effect of vertically aligned CNT arrays in the THz region, and paves a way for potential anisotropic THz applications based on CNTs with oblique incidence requirements.


Receptor-targeted lentiviral vectors are exceptionally sensitive toward the biophysical properties of the displayed single-chain Fv.
An increasing number of applications require the expression of single-chain variable fragments (scFv) fusion proteins in mammalian cells at the cell surface membrane. Here we assessed the CD30-specific scFv HRS3, which is used in immunotherapy, for its ability to retarget lentiviral vectors (LVs) to CD30 and to mediate selective gene transfer into CD30-positive cells. Fused to the C-terminus of the type-II transmembrane protein hemagglutinin (H) of measles virus and expressed in LV packaging cells, gene transfer mediated by the released LV particles was inefficient. A series of point mutations in the scFv framework regions addressing its biophysical properties, which substantially improved production and increased the melting temperature without impairing its kinetic binding behavior to CD30, also improved the performance of LV particles. Gene transfer into CD30-positive cells increased ∼100-fold due to improved transport of the H-scFv protein to the plasma membrane. Concomitantly, LV particle aggregation and syncytia formation in packaging cells were substantially reduced. The data suggest that syncytia formation can be triggered by trans-cellular dimerization of H-scFv proteins displayed on adjacent cells. Taken together, we show that the biophysical properties of the targeting ligand have a decisive role for the gene transfer efficiency of receptor-targeted LVs.


Bcl-XL inhibits membrane permeabilization by competing with Bax.
Although Bcl-XL and Bax are structurally similar, activated Bax forms large oligomers that permeabilize the outer mitochondrial membrane, thereby committing cells to apoptosis, whereas Bcl-XL inhibits this process. Two different models of Bcl-XL function have been proposed. In one, Bcl-XL binds to an activator, thereby preventing Bax activation. In the other, Bcl-XL binds directly to activated Bax. It has been difficult to sort out which interaction is important in cells, as all three proteins are present simultaneously. We examined the mechanism of Bax activation by tBid and its inhibition by Bcl-XL using full-length recombinant proteins and measuring permeabilization of liposomes and mitochondria in vitro. Our results demonstrate that Bcl-XL and Bax are functionally similar. Neither protein bound to membranes alone. However, the addition of tBid recruited molar excesses of either protein to membranes, indicating that tBid activates both pro- and antiapoptotic members of the Bcl-2 family. Bcl-XL competes with Bax for the activation of soluble, monomeric Bax through interaction with membranes, tBid, or t-Bid-activated Bax, thereby inhibiting Bax binding to membranes, oligomerization, and membrane permeabilization. Experiments in which individual interactions were abolished by mutagenesis indicate that both Bcl-XL-tBid and Bcl-XL-Bax binding contribute to the antiapoptotic function of Bcl-XL. By out-competing Bax for the interactions leading to membrane permeabilization, Bcl-XL ties up both tBid and Bax in nonproductive interactions and inhibits Bax binding to membranes. We propose that because Bcl-XL does not oligomerize it functions like a dominant-negative Bax in the membrane permeabilization process.


A pharmacometric approach to evaluate drugs for potential repurposing as COVID-19 therapeutics.
Developing and evaluating novel compounds for treatment or prophylaxis of emerging infectious diseases is costly and time-consuming. Repurposing of already available marketed compounds is an appealing option as they already have an established safety profile. This approach could substantially reduce cost and time required to make effective treatments available to fight the COVID-19 pandemic. However, this approach is challenging since many drug candidates show efficacy in in vitro experiments, but fail to deliver effect when evaluated in clinical trials. Better approaches to evaluate in vitro data are needed, in order to prioritize drugs for repurposing. This article evaluates potential drugs that might be of interest for repurposing in the treatment of patients with COVID-19 disease. A pharmacometric simulation-based approach was developed to evaluate in vitro activity data in combination with expected clinical drug exposure, in order to evaluate the likelihood of achieving effective concentrations in patients. The presented pharmacometric approach bridges in vitro activity data to clinically expected drug exposures, and could therefore be a useful compliment to other methods in order to prioritize repurposed drugs for evaluation in prospective randomized controlled clinical trials.


Long Noncoding RNA MALAT1 Promotes Aggressive Pancreatic Cancer Proliferation and Metastasis via the Stimulation of Autophagy.
Recently, pancreatic ductal adenocarcinoma (PDAC) has emerged as one of the most aggressive malignant tumors with the worst prognosis. Previous studies have demonstrated that long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is increased in pancreatic cancer and is identified as a diagnostic biomarker. Nonetheless, the molecular mechanism of elevated MALAT1 levels and tumor aggressiveness remains unknown. In this study, MALAT1 was found to be highly expressed in PDAC tissues, and elevated expression was associated with poorer prognoses. In addition, MALAT1 was positively linearly correlated with the expression of LC3B mRNA. Furthermore, several molecules involved in cellular autophagic flux were modulated following the downregulation of MALAT1, including LC3, P62, and LAMP-2. Mechanistically, we found that MALAT1 interacted with RNA binding protein HuR, and silencing of MALAT1 greatly enhanced the posttranscriptional regulation of TIA-1 and had further effects on inhibiting autophagy. MALAT1 was speculated to regulate tumorigenesis via HuR-TIA-1-mediated autophagic activation. Hence, we investigated the biological properties of MALAT1 in terms of tumor proliferation and metastasis by promoting autophagy in vitro In brief, these data demonstrate that MALAT1 could facilitate the advanced progression of tumors in vivo Our study highlights the new roles of MALAT1 on protumorigenic functioning and anticancer therapy via activating autophagy in pancreatic cancer. Mol Cancer Ther; 15(9); 2232-43. ©2016 AACR.


AT2 receptors: functional relevance in cardiovascular disease.
The renin angiotensin system (RAS) is intricately involved in normal cardiovascular homeostasis. Excessive stimulation by the octapeptide angiotensin II contributes to a range of cardiovascular pathologies and diseases via angiotensin type 1 receptor (AT1R) activation. On the other hand, tElsevier Inc.he angiotensin type 2 receptor (AT2R) is thought to counter-regulate AT1R function. In this review, we describe the enhanced expression and function of AT2R in various cardiovascular disease settings. In addition, we illustrate that the RAS consists of a family of angiotensin peptides that exert cardiovascular effects that are often distinct from those of Ang II. During cardiovascular disease, there is likely to be an increased functional importance of AT2R, stimulated by Ang II, or even shorter angiotensin peptide fragments, to limit AT1R-mediated overactivity and cardiovascular pathologies.


Rich Intrinsic Image Decomposition of Outdoor Scenes from Multiple Views.
Intrinsic images aim at separating an image into its reflectance and illumination components to facilitate further analysis or manipulation. This separation is severely ill posed and the most successful methods rely on user indications or precise geometry to resolve the ambiguities inherent to this problem. In this paper, we propose a method to estimate intrinsic images from multiple views of an outdoor scene without the need for precise geometry and with a few manual steps to calibrate the input. We use multiview stereo to automatically reconstruct a 3D point cloud of the scene. Although this point cloud is sparse and incomplete, we show that it provides the necessary information to compute plausible sky and indirect illumination at each 3D point. We then introduce an optimization method to estimate sun visibility over the point cloud. This algorithm compensates for the lack of accurate geometry and allows the extraction of precise shadows in the final image. We finally propagate the information computed over the sparse point cloud to every pixel in the photograph using image-guided propagation. Our propagation not only separates reflectance from illumination, but also decomposes the illumination into a sun, sky, and indirect layer. This rich decomposition allows novel image manipulations as demonstrated by our results.


Functional outcomes of Denis zone III sacral fractures treated nonoperatively.
The purpose of this study is to report the outcomes of nonoperative treatment in patients with Denis Zone III sacral fractures at a minimum of 2 years follow up. Retrospective review of prospectively collected data of a consecutive series of patients. Level I trauma center. A consecutive series of 15 patients (15-47 years old) with Denis Zone III sacral fractures treated nonoperatively from 1997 to 2002 was studied. Eleven patients were available for follow-up questionnaires; nine participated in a physical examination. Time to final follow up averaged 43 months (range, 25-67 months). Demographic data; mechanism of injury; injury-specific assessment of bowel, bladder, and sexual function; physical examination; and fracture pattern were collected from a prospectively collected database. At a minimum of 2-year follow up, evaluation of SF-36 scores, Roland Morris back pain questionnaire, and Gibbons classification was conducted. All fractures healed. Six patients had a postinjury increase in kyphosis (range, 1 degrees -17 degrees ) without a correlation to final outcomes. Mean SF-36 scores were all uniformly lower than the normalized general population and were biased by frequent associated injuries. Final Roland-Morris scores averaged 3.3 +/- 3.3. Gibbons classification scores initially averaged 2 +/- 1.2 and decreased to 1.5 +/- 0.8, each within their standard deviations. Eight had residual bowel, bladder, and/or sexual dysfunction. Nonoperative treatment of Denis Zone III sacral fractures yields consistent healing. Despite improvement in initial neurologic deficits, residual complaints were common.


Intraoperative cholangiogram during laparoscopic cholecystectomy: A clinical trial in rural setting.
The routine use of intraoperative cholangiogram (IOC) in laparoscopic cholecystectomy (LC) remains a contentious issue. IOC helps to delineate biliary tree anatomy, prevent bile duct injury and image stones in the common bile duct (CBD). It may prove to be a valuable alternative to ERCP or MRCP, especially in the rural setting with limited resources. All patients undergoing laparoscopic cholecystectomy during a 12-month period were audited. For the first 6 months, patients were recruited for routine IOC and for the second 6 months, routine IOC was not performed. Cases were analysed with regard to patient demographics, operative details and clinical outcomes. A total of 75 patients were analysed within the 12-month period. The majority were women aged 41-50. Ultrasound suggested common bile duct stones in 6.7% of cases. IOC was attempted in 50.7% of cases. Of these, 29 (76.3%) were successful. IOC added an average of 28 min to total theatre time. A total of 75% (n = 22) of IOCs showed normal flow of contrast into the intra- and extra-hepatic biliary systems. In 17% (n = 5) of patients, stones within the CBD were suspected, and these were referred for further management. ERCP/MRCP confirmed CBD stones in 60% (n = 3) of these patients. There was poor correlation between pre-operative suspicion and confirmed CBD stones (two patients only with pre-operative suspected CBD stone confirmed on IOC and ERCP). There were no operative complications related to IOC. There were no post-operative complications in cases where no IOC was done. The majority of patients treated in our centre were women, middle-aged patients booked for elective laparoscopic cholecystectomy. Although only 6.7% cases were suspicious for CBD stones pre-operatively, a total of 17% of patients with routine IOC suggested CBD stones. IOC was found to be safe, taking only an additional 28 min of total theatre time. Routine rather than selective use of IOC could be considered to improve patient safety and long-term results.


Osteomyelitis of the hyoid bones in two calves.
Two calves were referred because of ptyalism and difficulty opening the mouth (Calf 1) and for elective umbilical hernia surgery under inhalation anaesthesia (Calf 2). Additional clinical signs were increased breath sounds and swelling in the region of the mandibular angle in Calf 1. Ultrasonography and endoscopy revealed oral inflammation and abscessation in the area of the base of the tongue in both calves. Infection of the hyoid apparatus was suspected based on ultrasonographic findings and confirmed by means of computed tomography. In Calf 1, there was no response to treatment with systemic antibiotics, nonsteroidal anti-inflammatory drugs and local lavage, and Calf 2 was not treated. Both calves were euthanized because of a poor prognosis and the diagnoses confirmed during postmortem examination. In Calf 1, the abscess was associated with complete destruction of the left epihyoid bone and partial destruction of the left stylohyoid and ceratohyoid bones. In Calf 2, the abscess was located at the distal end of the right stylohyoid bone near the epihyoid bone. Stomatitis or laryngeal and pharyngeal abscessation caused by sharp feed particles are common in cattle and infection of the hyoid apparatus should be included in the differential diagnosis.


A method for the covalent capture and screening of diverse small molecules in a microarray format.
This protocol describes a robust method for the covalent capture of small molecules with diverse reactive functional groups in microarray format, and outlines a procedure for probing small-molecule microarrays (SMMs) with proteins of interest. A vapor-catalyzed, isocyanate-mediated surface immobilization scheme is used to attach bioactive small molecules, natural products and small molecules derived from diversity-oriented synthesis pathways. Additionally, an optimized methodology for screening SMMs with purified proteins and cellular lysates is described. Finally, a suggested model for data analysis that is compatible with commercially available software is provided. These procedures enable a platform capability for discovering novel interactions with potential applications to immunoglobulin profiling, comparative analysis of cellular states and ligand discovery. With the appropriate materials and experimental setup, the printing of SMMs can be completed in 14 hours over 3 days. Screening and data analysis requires 2 days. A detailed timeline is provided.


Mutant Cullin 3 causes familial hyperkalemic hypertension via dominant effects.
Mutations in the ubiquitin ligase scaffold protein Cullin 3 (CUL3) cause the disease familial hyperkalemic hypertension (FHHt). In the kidney, mutant CUL3 (CUL3-Δ9) increases abundance of With-No-Lysine [K] Kinase 4 (WNK4), with excessive activation of the downstream Sterile 20 (STE20)/SPS-1-related proline/alanine-rich kinase (SPAK) increasing phosphorylation of the Na+-Cl- cotransporter (NCC). CUL3-Δ9 promotes its own degradation via autoubiquitination, leading to the hypothesis that Cul3 haploinsufficiency causes FHHt. To directly test this, we generated Cul3 heterozygous mice (CUL3-Het), and Cul3 heterozygotes also expressing CUL3-Δ9 (CUL3-Het/Δ9), using an inducible renal epithelial-specific system. Endogenous CUL3 was reduced to 50% in both models, and consistent with autoubiquitination, CUL3-Δ9 protein was undetectable in CUL3-Het/Δ9 kidneys unless primary renal epithelia cells were cultured. Abundances of WNK4 and phosphorylated NCC did not differ between control and CUL3-Het mice, but they were elevated in CUL3-Het/Δ9 mice, which also displayed higher plasma [K+] and blood pressure. Abundance of phosphorylated Na+-K+-2Cl- cotransporter (NKCC2) was also increased, which may contribute to the severity of CUL3-Δ9-mediated FHHt. WNK4 and SPAK localized to puncta in NCC-positive segments but not in NKCC2-positive segments, suggesting differential effects of CUL3-Δ9. These results indicate that Cul3 haploinsufficiency does not cause FHHt, but dominant effects of CUL3-Δ9 are required.


Mechanistic understanding of molecular initiating events (MIEs) using NMR spectroscopy.
Toxicological risk assessments in the 21st century are increasingly being driven by the Adverse Outcome Pathways (AOP) conceptual framework in which the Molecular Initiating Event (MIE) is of fundamental importance to pathway progression. For those MIEs that involve covalent chemical reactions, such as protein haptenation, determination of relative rates and mechanisms of reactions is a prerequisite for their understanding. The utility of NMR spectroscopy as an experimental technique for effectively providing reaction rate and mechanistic information for early assessment of likely MIE(s) has been demonstrated. To demonstrate the concept, model systems exemplifying common chemical reactions involved in the covalent modification of proteins were utilized; these involved chemical reactions of electrophilic species (representing different mechanistic classes) with simple amine and thiol nucleophiles acting as surrogates for the reactive groups of lysine and cysteine protein side chains respectively. Such molecular interactions are recognized as critical mechanisms in a variety of chemical and drug toxicities, including respiratory and skin sensitization and liver toxicity as well as being the key mechanism of action for a number of therapeutic agents.


A novel approach to FRUC using discriminant saliency and frame segmentation.
Motion-compensated frame interpolation (MCFI) is a technique used extensively for increasing the temporal frequency of a video sequence. In order to obtain a high quality interpolation, the motion field between frames must be well-estimated. However, many current techniques for determining the motion are prone to errors in occlusion regions, as well as regions with repetitive structure. We propose an algorithm for improving both the objective and subjective quality of MCFI by refining the motion vector field. We first utilize a discriminant saliency classifier to determine which regions of the motion field are most important to a human observer. These regions are refined using a multistage motion vector refinement (MVR), which promotes motion vector candidates based upon their likelihood given a local neighborhood. For regions which fall below the saliency-threshold, a frame segmentation is used to locate regions of homogeneous color and texture via normalized cuts. Motion vectors are promoted such that each homogeneous region has a consistent motion. Experimental results demonstrate an improvement over previous frame rate up-conversion (FRUC) methods in both objective and subjective picture quality.


Identifying thresholds for classifying childhood psychiatric disorder: issues and prospects.
To evaluate empirically the implications of choosing different thresholds to classify conduct disorder and attention-deficit hyperactivity disorder for estimating prevalence, test-retest reliability of measurement, and informant (parent/teacher) agreement and for evaluating comorbidity and associated features of disorder. Data for the study came from problem checklist assessments done by parents and teachers of children aged 6 to 16 years (N = 1,229) selected with known probability from a general population sample and from structured interviews obtained in a stratified, random subsample (n = 251). Estimates varied widely depending on the rationale used to set thresholds. Percent prevalence went from 0.1 to 39.2; kappa estimates of test-retest reliability went from .19 to .82. Parent-teacher agreement based on kappa went from .0 to .38. Relative odds between disorder and associated features varied twofold. Use of different rationales to set thresholds for classifying childhood psychiatric disorder in the general population has profound implications for what we learn about the epidemiology of childhood disorder.


Epidemiology and susceptibilities of methicillin-resistant Staphylococcus aureus in Northeastern Ohio.
A retrospective survey was performed on all staphylococcal infections diagnosed by the Ashtabula County Medical Center (Ashtabula, OH) during 2006 and 2007. Of the 1,612 Staphylococcus aureus isolates evaluated for their antibiotic resistances, 947 were methicillin-resistant S aureus (MRSA). In 2007, MRSA infections reached 589 cases per 100,000 inhabitants, a 77% increase compared with 2006. The increase in MRSA infections was noticeable among youth (6-25 years old), middle-aged people (45-50 years old), and elderly people (86-90 years old). MRSA infections increased among inpatients by 58%, among outpatients by 43%, and among nursing home residents by 183%. More than 66% of MRSA infections were found among healthy people in the community with no apparent risk factors. More than 88.7% of the infections belong to only 9 profiles of antibiotic resistance indiscriminately distributed among inpatients, outpatients, and nursing home residents. This report sheds further light on the rapid spread of MRSA across Northeastern Ohio, stressing the need for better education in preventive measures and infection control at the level of community and health care settings.


Purification and characterization of a GTP-pyrophosphate exchange activity from vaccinia virions. Association of the GTP-pyrophosphate exchange activity with vaccinia mRNA guanylyltransferase . RNA (guanine-7-)methyltransferase complex (capping enzyme).
A core-associated enzyme, which catalyzes a nucleotide-pyrophosphate exchange with GTP, has been purified from vaccinia virions. The enzyme requires MgCl2 for activity, has an alkaline pH optimum, and specifically utilizes GTP as the exchanging nucleotide. The enzyme does not catalyze exchange of GMP with GTP. The GTP-PPi exchange enzyme co-purifies with vaccinia capping enzyme (RNA guanylyltransferase and RNA (guanine-7-)methyltransferase) through successive chromatography steps on DEAE-cellulose, DNA-cellulose, and phosphocellulose. GTP-PPi exchange and capping activities remain physically associated during sedimentation in a glycerol gradient. Under high salt conditions (1 M NaCl), GTP-PPi exchange, capping, and methylating activities co-sediment with an RNA triphosphatase activity and a nucleoside triphosphate phosphohydrolase activity as a 6.5 S multifunctional enzyme complex which contains two major polypeptides of 96,000 and 26,000 molecular weight. The characteristics of the various enzymatic reactions catalyzed by this complex are described. The GTP-PPi exchange reaction of vaccinia guanylyltransferase affords a simple, sensitive assay for capping enzyme function. The relevance of the GTP-PPi exchange reaction to the mechanism of transguanylylation is considered.


Deformable and rigid registration of MRI and microPET images for photodynamic therapy of cancer in mice.
We are investigating imaging techniques to study the tumor response to photodynamic therapy (PDT). Positron emission tomography (PET) can provide physiological and functional information. High-resolution magnetic resonance imaging (MRI) can provide anatomical and morphological changes. Image registration can combine MRI and PET images for improved tumor monitoring. In this study, we acquired high-resolution MRI and microPET 18F-fluorodeoxyglucose (FDG) images from C3H mice with RIF-1 tumors that were treated with Pc 4-based PDT. We developed two registration methods for this application. For registration of the whole mouse body, we used an automatic three-dimensional, normalized mutual information algorithm. For tumor registration, we developed a finite element model (FEM)-based deformable registration scheme. To assess the quality of whole body registration, we performed slice-by-slice review of both image volumes; manually segmented feature organs, such as the left and right kidneys and the bladder, in each slice; and computed the distance between corresponding centroids. Over 40 volume registration experiments were performed with MRI and microPET images. The distance between corresponding centroids of organs was 1.5 +/- 0.4 mm which is about 2 pixels of microPET images. The mean volume overlap ratios for tumors were 94.7% and 86.3% for the deformable and rigid registration methods, respectively. Registration of high-resolution MRI and microPET images combines anatomical and functional information of the tumors and provides a useful tool for evaluating photodynamic therapy.


Dynamic CD138 surface expression regulates switch between myeloma growth and dissemination.
The canonical plasma cell marker CD138 (syndecan-1) is highly expressed on the myeloma cell surface, but its functional role in vivo is unclear, as well as the ontogeny of CD138-high and CD138-negative (neg) myeloma cells. In this study we used an in vivo murine Vk*MYC myeloma model where CD138 is heterogeneously expressed depending on tumor size. We find that in comparison to CD138-neg myeloma cells, the CD138-high subset of myeloma cells is highly proliferative, less apoptotic, and enhanced IL-6R signaling, which is known to promote survival. In addition CD138-high myeloma engrafts better than its CD138-neg counterpart. In contrast, CD138-neg cells are more motile both in vitro and in vivo, and more readily disseminate and spread to other bones in vivo than CD138-high subset. Neutralizing CD138 rapidly triggers migration of myeloma cells in vivo and leads to intravasation, which results in increased dissemination to other bones. Both murine and human myeloma cells can rapidly recycle CD138 surface expression through endocytic trafficking, in response to serum levels. Blocking CD138 enhances myeloma sensitivity to bortezomib chemotherapy and significantly reduces tumor size compared to bortezomib treatment alone. Thus, our data show that CD138 surface expression dynamically regulates a switch between growth vs. dissemination for myeloma, in response to nutrient conditions.


Clinical and laboratory profile of febrile respiratory infections in general practice.
Respiratory tract infection (RTI) is a common diagnosis in the primary care setting. The aim of the study was to characterize the clinical course and laboratory manifestations of febrile RTI in Israeli adults presenting to primary care clinics. A prospective study over a 3-month winter period in 3 urban university primary care clinics of 122 consecutive adult patients seen by their family practitioners. All participants were interviewed and had chest radiographs, tests of oxygen saturation, and blood tests. Study group included 122 adults (mean age, 44.8 +/- 14.2 years; men, 38%). Fever lasted for a mean of 4.0 +/- 1.7 days, and the mean number of days off work/activities was 5.3 +/- 1.0. Pneumonia was clinically suspected in 33 patients (27%), but documented in 7 (6%). Antibiotics were given to 94 patients (77%). The group of treated patients had a lower mean oxygen saturation (P <.05) and a higher erythrocyte sedimentation rate (P <.05). Blood tests were found to be abnormal, although not clinically significant, in some patients, and all resolved spontaneously. RTIs are usually of short duration with a benign course where laboratory blood tests have no clinical implication. Radiographs may distinguish the 6% of patients having pneumonia from the 27% with suspected cases.


Gambling disorders.
Gambling disorders, including pathological gambling and problem gambling, have received increased attention from clinicians and researchers over the past three decades since gambling opportunities have expanded around the world. This Seminar reviews prevalence, causes and associated features, screening and diagnosis, and treatment approaches. Gambling disorders affect 0·2-5·3% of adults worldwide, although measurement and prevalence varies according to the screening instruments and methods used, and availability and accessibility of gambling opportunities. Several distinct treatment approaches have been favourably evaluated, such as cognitive behavioural and brief treatment models and pharmacological interventions. Although promising, family therapy and support from Gamblers Anonymous are less well empirically supported. Gambling disorders are highly comorbid with other mental health and substance use disorders, and a further understanding is needed of both the causes and treatment implications of this disorder.


[Evaluation of P-glycoprotein mediated in vitro loperamide biliary excretion with sandwich-cultured rat hepatocytes model].
An in vitro P-glycoprotein mediated drug biliary excretion model (B-Clear model) was developed and validated using sandwich-cultured rat hepatocytes (SCRH) and a model substrate rhodamine 123 (Rh123). SCRH formed functional bile canalicular networks after 5 days of culture. Rh123 (10 micromol x L(-1)) was then incubated with the SCRH in standard Ca+ Hanks buffer or Ca(2+)-free buffer. The cumulative cell uptake and canalicular efflux of Rh123 under Ca2+ and Ca(2+)-free conditions were measured with a LC-MS/MS method. The biliary excretion index (BEI) and instinct biliary clearance (CL(bile, int)) were calculated. To assess the effect of known P-gp inhibitors on the efflux of Rh123, cyclosporine A (CyA), tariquidar (TQD) or quinidine (QND) (10, 50 and 100 micromol x L(-1)) was pre-incubated separately with SCRH for 30 min, then co-incubated with Rh123. The BEI and CL(bile, int) of Rh123 obtained from the SCRH model were (17.8 +/- 1.3) % and (10.7 +/- 0.9) mL x min(-1) x kg(-1), respectively. All the three P-gp inhibitors showed a dose-dependent inhibition on the bile clearance of Rh123, indicating that the B-Clear model with SCRH was functional properly. The biliary excretion of loperamide (LPAD) and the role of P-gp were further investigated with this validated model. The BEI and CL(bile, int) for LPAD (20 micromol x L(-1)) were obtained after it was incubated with SCRH for 30 min, and found to be (12.9 +/- 1.2)% and (6.1 +/- 0.3) mL x min(-1) x kg(-1) respectively. The dose-dependent inhibition on LPAD biliary excretion by CyA, TQD or QND confirmed the major role of P-gp in LPAD canalicular efflux. The results suggested that the B-Clear model with SCRH would be a useful tool for evaluation of P-gp mediated efflux and drug-drug interaction.


Variants of uncertain significance in BRCA: a harbinger of ethical and policy issues to come?
After two decades of genetic testing and research, the BRCA1 and BRCA2 genes are two of the most well-characterized genes in the human genome. As a result, variants of uncertain significance (VUS; also called variants of unknown significance) are reported less frequently than for genes that have been less thoroughly studied. However, VUS continue to be uncovered, even for BRCA1/2. The increasing use of multi-gene panels and whole-genome and whole-exome sequencing will lead to higher rates of VUS detection because more genes are being tested, and most genomic loci have been far less intensively characterized than BRCA1/2. In this article, we draw attention to ethical and policy-related issues that will emerge. Experience garnered from BRCA1/2 testing is a useful introduction to the challenges of detecting VUS in other genetic testing contexts, while features unique to BRCA1/2 suggest key differences between the BRCA experience and the current challenges of multi-gene panels in clinical care. We propose lines of research and policy development, emphasizing the importance of pooling data into a centralized open-access database for the storage of gene variants to improve VUS interpretation. In addition, establishing ethical norms and regulated practices for sharing and curating data, analytical algorithms, interpretive frameworks and patient re-contact are important policy areas.


Closed-Loop Feedback Illumination for Optical Inverse Tone-Mapping in Light Microscopy.
In this paper, we show that optical inverse tone-mapping (OITM) in light microscopy can improve the visibility of specimens, both when observed directly through the oculars and when imaged with a camera. In contrast to previous microscopy techniques, we premodulate the illumination based on the local modulation properties of the specimen itself. We explain how the modulation of uniform white light by a specimen can be estimated in real time, even though the specimen is continuously but not uniformly illuminated. This information is processed and back-projected constantly, allowing the illumination to be adjusted on the fly if the specimen is moved or the focus or magnification of the microscope is changed. The contrast of the specimen's optical image can be enhanced, and high-intensity highlights can be suppressed. A formal pilot study with users indicates that this optimizes the visibility of spatial structures when observed through the oculars. We also demonstrate that the signal-to-noise (S/N) ratio in digital images of the specimen is higher if captured under an optimized rather than a uniform illumination. In contrast to advanced scanning techniques that maximize the S/N ratio using multiple measurements, our approach is fast because it requires only two images. This can improve image analysis in digital microscopy applications with real-time capturing requirements.


Analysis of viral testing in nonacetaminophen pediatric acute liver failure.
Viral infections are often suspected to cause pediatric acute liver failure (PALF), but large-scale studies have not been performed. We analyzed the results of viral testing among nonacetaminophen PALF study participants. Participants were enrolled in the PALF registry. Diagnostic evaluation and final diagnosis were determined by the site investigator and methods for viral testing by local standard of care. Viruses were classified as either causative viruses (CVs) or associated viruses (AVs). Supplemental testing for CV was performed if not done clinically and serum was available. Final diagnoses included "viral," "indeterminate," and "other." Of 860 participants, 820 had at least 1 test result for a CV or AV. A positive viral test was found in 166/820 (20.2%) participants and distributed among "viral" (66/80 [82.5%]), "indeterminate" (52/420 [12.4%]), and "other" (48/320 [15.0%]) diagnoses. CVs accounted for 81/166 (48.8%) positive tests. Herpes simplex virus (HSV) was positive in 39/335 (11.6%) who were tested 26/103 (25.2%) and 13/232 (5.6%) among infants 0 to 6 and >6 months, respectively. HSV was not tested in 61.0% and 53% of the overall cohort and those 0 to 6 months, respectively. Supplemental testing yielded 17 positive, including 5 HSV. Viral testing in PALF occurs frequently but is often incomplete. The evidence for acute viral infection was found in 20.2% of those tested for viruses. HSV is an important viral cause for PALF in all age groups. The etiopathogenic role of CV and AV in PALF requires further investigation.


A structured simple form for ordering genetic tests is needed to ensure coupling of clinical detail (phenotype) with DNA variants (genotype) to ensure utility in publication and databases.
Researchers and clinicians ideally need instant access to all the variation in their gene/locus of interest to efficiently conduct their research and genetic healthcare to the highest standards. Currently much key data resides in the laboratory books or patient records around the world, as there are many impediments to submitting this data. It would be ideal therefore if a semiautomated pathway was available, with a minimum of effort, to make the deidentified data publicly available for others to use. The Human Variome Project (HVP) meeting listed 96 recommendations to work toward this situation. This article is planned to initiate a strategy to enhance the collection of phenotype and genotype data from the clinician/diagnostic laboratory nexus. Thus, the aim is to develop universally applicable forms that people can use when investigating patients for each inherited disease, to assist in satisfying many of the recommendations of the HVP Meeting [Cotton et al., 2007]. We call for comment and collaboration in this article.