Peripheral nerve injury increases glutamate-evoked calcium mobilization in adult spinal cord neurons.
Central sensitization in the spinal cord requires glutamate receptor activation and intracellular Ca2+ mobilization. We used Fura-2 AM bulk loading of mouse slices together with wide-field Ca2+ imaging to measure glutamate-evoked increases in extracellular Ca2+ to test the hypotheses that: 1. Exogenous application of glutamate causes Ca2+ mobilization in a preponderance of dorsal horn neurons within spinal cord slices taken from adult mice; 2. Glutamate-evoked Ca2+ mobilization is associated with spontaneous and/or evoked action potentials; 3. Glutamate acts at glutamate receptor subtypes to evoked Ca2+ transients; and 4. The magnitude of glutamate-evoked Ca2+ responses increases in the setting of peripheral neuropathic pain. Bath-applied glutamate robustly increased [Ca2+]i in 14.4 ± 2.6 cells per dorsal horn within a 440 x 330 um field-of-view, with an average time-to-peak of 27 s and decay of 112 s. Repeated application produced sequential responses of similar magnitude, indicating the absence of sensitization, desensitization or tachyphylaxis. Ca2+ transients were glutamate concentration-dependent with a Kd = 0.64 mM. Ca2+ responses predominantly occurred on neurons since: 1) Over 95% of glutamate-responsive cells did not label with the astrocyte marker, SR-101; 2) 62% of fura-2 AM loaded cells exhibited spontaneous action potentials; 3) 75% of cells that responded to locally-applied glutamate with a rise in [Ca2+]i also showed a significant increase in AP frequency upon a subsequent glutamate exposure; 4) In experiments using simultaneous on-cell recordings and Ca2+ imaging, glutamate elicited a Ca2+ response and an increase in AP frequency. AMPA/kainate (CNQX)- and AMPA (GYKI 52466)-selective receptor antagonists significantly attenuated glutamate-evoked increases in [Ca2+]i, while NMDA (AP-5), kainate (UBP-301) and class I mGluRs (AIDA) did not. Compared to sham controls, peripheral nerve injury significantly decreased mechanical paw withdrawal threshold and increased glutamate-evoked Ca2+ signals. Bulk-loading fura-2 AM into spinal cord slices is a successful means for determining glutamate-evoked Ca2+ mobilization in naïve adult dorsal horn neurons. AMPA receptors mediate the majority of these responses. Peripheral neuropathic injury potentiates Ca2+ signaling in dorsal horn.


Acitretin therapy is effective for psoriasis associated with human immunodeficiency virus infection.
To determine the safety, tolerability, and effectiveness of a newer retinoid, acitretin, as monotherapy for psoriasis associated with human immunodeficiency virus infection (PS-HIV). Pilot investigation. An academic medical center. Eleven patients selected from volunteers with PS-HIV were enrolled in a 20-week treatment protocol. Two patients discontinued participation in the study because of worsening psoriasis; a third patient was unable to continue treatment after having a myocardial infarction, presumably unrelated to acitretin therapy. Each patient received an optimized dose of acitretin during the period of observation. Clinical and laboratory assessments were performed every 2 weeks during the trial. The Psoriasis Area and Severity Index was used to assess the clinical response to treatment. To monitor for toxic drug effects, a panel of laboratory parameters, including complete blood cell count, biochemistry profile, urinalysis, HLA typing, skin biopsy for histological examination, and T-cell counts, was performed. Six (54%) of 11 patients with PS-HIV achieved good to excellent responses using acitretin monotherapy. Four patients (36%) achieved complete clearing. There was no evidence of a correlation between the pretreatment measures of immunosuppression and the therapeutic response. Parameters of immunosuppression were not exacerbated by acitretin therapy. Acitretin is a safe and effective treatment for PS-HIV. Both skin and joint manifestations of PS-HIV responded to acitretin therapy in most patients. Optimal results were achieved with a dose of 75 mg/d. The adverse effects were moderate and well tolerated. Acitretin does not appear to have immunosuppressive properties. A formal randomized clinical trial is warranted.


The Sec62-Sec63 translocon facilitates translocation of the C-terminus of membrane proteins.
The Sec62-Sec63 complex mediates post-translational translocation of a subset of primarily secretory proteins into the endoplasmic reticulum (ER) in yeast. Therefore, it has been thought that membrane proteins, which are mainly co-translationally targeted into the ER, are not handled by the Sec62-Sec63 translocon. By systematic analysis of single and multi-spanning membrane proteins with broad sequence context [with differing hydrophobicity, flanking charged residues and orientation of transmembrane (TM) segments], we show that mutations in the N-terminal cytosolic domain of yeast Sec62 impair its interaction with Sec63 and lead to defects in membrane insertion and translocation of the C-terminus of membrane proteins. These results suggest that there is an unappreciated function of the Sec62-Sec63 translocon in regulating topogenesis of membrane proteins in the eukaryotic cell.


A gradient-directed Monte Carlo method for global optimization in a discrete space: application to protein sequence design and folding.
We apply the gradient-directed Monte Carlo (GDMC) method to select optimal members of a discrete space, the space of chemically viable proteins described by a model Hamiltonian. In contrast to conventional Monte Carlo approaches, our GDMC method uses local property gradients with respect to chemical variables that have discrete values in the actual systems, e.g., residue types in a protein sequence. The local property gradients are obtained from the interpolation of discrete property values, following the linear combination of atomic potentials scheme developed recently [M. Wang et al., J. Am. Chem. Soc. 128, 3228 (2006)]. The local property derivative information directs the search toward the global minima while the Metropolis criterion incorporated in the method overcomes barriers between local minima. Using the simple HP lattice model, we apply the GDMC method to protein sequence design and folding. The GDMC algorithm proves to be particularly efficient, suggesting that this strategy can be extended to other discrete optimization problems in addition to inverse molecular design.


Substantial effects of epimuscular myofascial force transmission on muscular mechanics have major implications on spastic muscle and remedial surgery.
The specific aim of this paper is to review the effects of epimuscular myofascial force transmission on muscular mechanics and present some new results on finite element modeling of non-isolated aponeurotomized muscle in order to discuss the dependency of mechanics of spastic muscle, as well as surgery for restoration of function on such force transmission. The etiology of the effects of spasticity on muscular mechanics is not fully understood. Clinically, such effects feature typically a limited joint range of motion, which at the muscle level must originate from altered muscle length-force characteristics, in particular a limited muscle length range of force exertion. In studies performed to understand what is different in spastic muscle and what the effects of remedial surgery are, muscle is considered as being independent of its surroundings. Conceivably, this is because the classical approach in muscle mechanics is built on experimenting with dissected muscles. Certainly, such approach allowed improving our understanding of fundamental muscle physiology yet it yielded implicitly a narrow point of view of considering muscle length-force characteristics as a fixed property of the muscle itself. However, within its context of its intact connective tissue surroundings (the in vivo condition) muscle is not an isolated and independent entity. Instead, collagenous linkages between epimysia of adjacent muscles provide direct intermuscular connections, and structures such as the neurovascular tracts provide indirect intermuscular connections. Moreover, compartmental boundaries (e.g., intermuscular septa, interosseal membranes, periost and compartmental fascia) are continuous with neurovascular tracts and connect muscular and non-muscular tissues at several locations additional to the tendon origins and insertions. Epimuscular myofascial force transmission occurring via this integral system of connections has major effects on muscular mechanics including substantial proximo-distal force differences, sizable changes in the determinants of muscle length-force characteristics (e.g. a condition dependent shift in muscle optimum length to a different length or variable muscle optimal force) explained by major serial and parallel distributions of sarcomere lengths. Therefore, due to epimuscular myofascial force transmission, muscle length-force characteristics are variable and muscle length range of force exertion cannot be considered as a fixed property of the muscle. The findings reviewed presently show that acutely, the mechanical mechanisms manipulated in remedial surgery are dominated by epimuscular myofascial force transmission. Conceivably, this is also true for the mechanism of adaptation during and after recovery from surgery. Moreover, stiffened epimuscular connections and therefore a stiffened integral system of intra- and epimuscular myofascial force transmission are indicated to affect the properties of spastic muscle. We suggest that important advancements in our present understanding of such properties, variability in the outcome of surgery and considerable recurrence of the impeded function after recovery cannot be made without taking into account the effects of epimuscular myofascial force transmission.


Three-Dimensional Imaging in Orthodontics.
Orthodontic records are one of the main milestones in orthodontic therapy. Records are essential not only for diagnosis and treatment planning but also for follow-up of the case, communicating with colleagues, and evaluating the treatment outcomes. Recently, two-dimensional (2D) imaging technology, such as cephalometric and panoramic radiographs and photographs, and plaster models were routinely used. However, after the introduction of three-dimensional (3D) technologies (laser scanner, stereophotogrammetry, and computed tomography) into dentistry, 3D imaging systems are more and more commonly preferred than 2D, especially in cases with craniofacial deformities. In fact, 3D imaging provided more detailed and realistic diagnostic information about the craniofacial hard as well as soft tissue and allowed to perform easier, faster, and more reliable 3D analyses. The purpose of this review is to provide an overview of the 3D imaging techniques, including their advantages and disadvantages, and to outline the indications for 3D imaging.


Orientin improves depression-like behavior and BDNF in chronic stressed mice.
Oxidative stress is involved in chronic stress-induced depression and the disruption of neurotransmission and neuroplasticity. Recently, orientin, a phenolic compound abundant in some fruits, millet, and herbs, has been shown to have antioxidant properties. This study investigated the potential antidepressant effects of orientin against chronic stress and its underlying mechanisms. The chronic unpredictable mild stress (CUMS) model was used to investigate the effects of orientin on behavior and biochemical alterations in mice. After 2 weeks of the CUMS protocol, the mice were treated with orientin (20 mg/kg and 40 mg/kg, oral gavage) for 3 weeks. Administration of orientin significantly alleviated the CUMS-induced depression-like behavior, including sucrose preference reduction, locomotor activity decline, and hypomotility. Orientin treatment attenuated the oxidative stress markers and increased the concentrations of serotonin and norepinephrine in the hippocampus and prefrontal cortex of CUMS mice. Orientin treatment also increased the brain-derived neurotrophic factor and synapse-associated proteins (synaptophysin and postsynaptic density protein 95) of CUMS mice. Orientin exerts antidepressant-like effects on CUMS mice, specifically by improving central oxidative stress, neurotransmission, and neuroplasticity. Therefore, supplementation with orientin-enriched food or fruit could be beneficial as a preventive strategy for chronic stress-induced depression.


First percutaneous Micra leadless pacemaker implantation and tricuspid valve repair with MitraClip NT for lead-associated severe tricuspid regurgitation.
Pacemaker lead-associated severe tricuspid regurgitation (TR) can lead to right heart failure and poor prognosis. Surgery in these patients carries significant morbidities. We describe a successful treatment of symptomatic severe TR by leadless pacemaker implantation followed by tricuspid valve (TV) repair with the MitraClip NT. A 71-year-old frail female with poor functional status, chronic atrial fibrillation and permanent pacemaker implantation in 2012 presented with symptomatic moderate-severe mitral regurgitation (MR) and severe TR with the pacemaker lead as the culprit. She was deemed extreme risk for double valve surgery and, because of her pacemaker dependency, the decision was to stage her interventions first with transcatheter mitral repair, then laser lead extraction and leadless pacemaker implantation to free the TV from tethering, then TV repair. An obstructive LAD lesion was identified and treated during mitral repair with the MitraClip NT. The Micra leadless pacemaker implantation and subsequent TV repair with the MitraClip NT were successful and the patient's MR improved to mild and TR to moderate, respectively. We report here a first successful transcatheter strategy to treat lead-associated severe TR by leadless pacemaker and MitraClip. Removing the pacemaker lead relieved leaflet tethering and improved the reparability of the TV.


Factors involved in hepatic glutathione depletion induced by acute ethanol administration.
Factors implicated in changes of the hepatic glutathione concentration following acute ethanol administration were examined in rats. Adult female rats were treated with either ethanol (4 g/kg, p.o.) or an isocaloric glucose solution. The hepatic reduced glutathione (GSH) concentration decreased rapidly after ethanol intake with a maximum diminution, approximately 50% of the control value, being observed at t = 6 h. The hepatic GSH concentration gradually increased, and finally rebounded at 24 h after ethanol ingestion. The dose of ethanol induced a transient increase in the oxidized glutathione (GSSG)/GSH ratio, which was associated with a significant reduction in GSH rather than elevation in GSSG [corrected]. The activity of gamma-glutamylcysteine synthetase (GCS), the rate-limiting enzyme for glutathione synthesis, and the cysteine concentration in liver were also measured. The GCS activity was depressed to approximately 80% of the control value at t = 2.5 h followed by rapid recovery, but no difference in the hepatic cysteine concentration between control and ethanol treated rats was observed for 24 h, suggesting that the reduction in glutathione synthesis may not play a major role in the significant depletion of this tripeptide in liver. The total glutathione concentration was measured both in prehepatic and posthepatic inferior vena cava blood. The glutathione concentration in posthepatic blood was approximately twice as high as that of prehepatic blood in control rats. Acute ethanol administration doubled the elevation of glutathione in posthepatic blood measured at t = 2.5 h. The sinusoidal efflux of glutathione estimated from the increase in blood glutathione concentration was greater than the total amount of its depletion in the liver of rats treated with ethanol. The results suggest that in the liver of rats treated acutely with ethanol, glutathione efflux plays the most important role in the reduction of this tripeptide, which would be aggravated by a transient decrease in glutathione synthesis and by increased consumption in association with its metabolism.


Festival foods in the immigrant diet.
Dietary acculturation for immigrant groups has largely been attributed to the "Westernization" of indigenous diets, as characterized by an increased consumption of unhealthy American foods (i.e., fast foods, hamburgers). However, acculturation and adoption of western dietary habits may not fully explain new dietary patterns among racial/ethnic minority immigrants. The immigrant diet may change in such a way that it elaborates on specific ethnic traditions in addition to the incorporation of Western food habits. In this paper, we explore the role that festival foods, those foods that were once eaten a few times a year and on special occasions, play in the regular diet of immigrants to the US. This paper will focus on the overconsumption of ethnic festival foods, which are often high in carbohydrates, animal protein, sugar and fat, as opposed to Western "junk" food, as an explanation for the increased risk of cardiometabolic disorders among new immigrant groups.


Metabolic impact of sex hormones on obesity.
Obesity and its associated health disorders and costs are increasing. Men and post-menopausal women have greater risk of developing complications of obesity than younger women. Within the brain, the hypothalamus is an important regulator of energy homeostasis. Two of its sub-areas, the ventrolateral portion of the ventral medial nucleus (VL VMN) and the arcuate (ARC) respond to hormones and other signals to control energy intake and expenditure. When large lesions are made in the hypothalamus which includes both the VL VMN and the ARC, animals eat more, have reduced energy expenditure, and become obese. The ARC and the VL VMN, in addition to other regions in the hypothalamus, have been demonstrated to contain estrogen receptors. There are two estrogen receptors, estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta). We and others have previously demonstrated that activation of ERalpha by estrogens reduces food intake and increases body weight. This review focuses on the relative contribution of activation of ERalpha by estrogens in the ARC and the VL VMN in the regulation of food intake and body weight. Additionally, estrogen receptors have been found in many peripheral tissues including adipose tissue. Estrogens are thought to have direct effects on adipose tissue and estrogens may provide anti-inflammatory properties both in the periphery and the in the central nervous system (CNS) which may protect women from diseases associated with inflammation. Understanding the mechanisms by which estrogens regulate body weight and inflammation will assist in determining potential therapeutic agents for menopausal women to decrease the propensity of diseases associated with obesity.


Understanding the effects of past flood events and perceived and estimated flood risks on individuals' voluntary flood insurance purchase behavior.
Over the past several decades, the economic damage from flooding in the coastal areas has greatly increased due to rapid coastal development coupled with possible climate change impacts. One effective way to mitigate excessive economic losses from flooding is to purchase flood insurance. Only a minority of coastal residents however have taken this preventive measure. Using original survey data for all coastal counties of the United States Gulf Coast merged with contextual data, this study examines the effects of external influences and perceptions of flood-related risks on individuals' voluntary behaviors to purchase flood insurance. It is found that the estimated flood hazard conveyed through the U.S. Federal Emergency Management Agency's (FEMA's) flood maps, the intensities and consequences of past storms and flooding events, and perceived flood-related risks significantly affect individual's voluntary purchase of flood insurance. This behavior is also influenced by home ownership, trust in local government, education, and income. These findings have several important policy implications. First, FEMA's flood maps have been effective in conveying local flood risks to coastal residents, and correspondingly influencing their decisions to voluntarily seek flood insurance in the U.S. Gulf Coast. Flood maps therefore should be updated frequently to reflect timely and accurate information about flood hazards. Second, policy makers should design strategies to increase homeowners' trust in the local government, to better communicate flood risks with residents, to address the affordability issue for the low-income, and better inform less educated homeowners through various educational programs. Future studies should examine the voluntary flood insurance behavior across countries that are vulnerable to flooding.


Production of the Alzheimer amyloid beta protein by normal proteolytic processing.
The 4-kilodalton (39 to 43 amino acids) amyloid beta protein (beta AP), which is deposited as amyloid in the brains of patients with Alzheimer's diseases, is derived from a large protein, the amyloid beta protein precursor (beta APP). Human mononuclear leukemic (K562) cells expressing a beta AP-bearing, carboxyl-terminal beta APP derivative released significant amounts of a soluble 4-kilodalton beta APP derivative essentially identical to the beta AP deposited in Alzheimer's disease. Human neuroblastoma (M17) cells transfected with constructs expressing full-length beta APP and M17 cells expressing only endogenous beta APP also released soluble 4-kilodalton beta AP, and a similar, if not identical, fragment was readily detected in cerebrospinal fluid from individuals with Alzheimer's disease and normal individuals. Thus cells normally produce and release soluble 4-kilodalton beta AP that is essentially identical to the 4-kilodalton beta AP deposited as insoluble amyloid fibrils in Alzheimer's disease.


The Analysis of 13C/12C Ratios in Exhaled CO2: Its Advantages and Potential Application to Field Research to Infer Diet, Changes in Diet Over Time, and Substrate Metabolism in Birds.
Stable isotopes are becoming an increasingly powerful tool for studying the physiological ecology of animals. The (13)C/(12)C ratios of animal tissues are frequently used to reconstruct the diet of animals. This usually requires killing the subjects. While there is an extensive medical literature on measuring the (13)C/(12)C ratio of exhaled CO(2) to determine substrate digestion and oxidation, we found little evidence that animal physiologists or physiological ecologists have applied (13)C/(12)C breath analysis in their studies. The analysis breath (13)C/(12)C ratios has the advantage of being non-invasive and non-destructive and can be repeatedly used on the same individual. Herein we briefly discuss the medical literature. We then discuss research which shows that, not only can the breath(13)C/(12)C ratio indicate what an animal is currently eating, but also the animal's diet in the past, and any changes in diet have occurred over time. We show that naturally occurring (13)C/(12)C ratios in exhaled CO(2) provides quantitative measure of the relative contribution of carbohydrates and lipids to flight metabolism. This technique is ripe for application to field research, and we encourage physiological ecologists to add this technique to their toolbox.


Clonal expansion of the Belgian Phytophthora ramorum populations based on new microsatellite markers.
Co-existence of both mating types A1 and A2 within the EU1 lineage of Phytophthora ramorum has only been observed in Belgium, which begs the question whether sexual reproduction is occurring. A collection of 411 Belgian P. ramorum isolates was established during a 7-year survey. Our main objectives were genetic characterization of this population to test for sexual reproduction, determination of population structure, evolution and spread, and evaluation of the effectiveness and impact of control measures. Novel, polymorphic simple sequence repeat (SSR) markers were developed after screening 149 candidate loci. Eighty isolates of P. ramorum, broadly representing the Belgian population, were analyzed using four previously described and three newly identified polymorphic microsatellite loci as well as amplified fragment length polymorphisms. SSR analysis was most informative and was used to screen the entire Belgian population. Thirty multilocus genotypes were identified, but 68% of the isolates belonged to the main genotype EU1MG1. Although accumulated mutation events were detected, the overall level of genetic diversity within the Belgian isolates of P. ramorum appears to be limited, indicating a relatively recent clonal expansion. Based on our SSR analysis there is no evidence of sexual recombination in the Belgian population of P. ramorum. Metalaxyl use decreased the genetic diversity of P. ramorum until 2005, when the majority of the isolates had become resistant. Most genotypes were site-specific and despite systematic removal of symptomatic and neighbouring plants, some genotypes were detected over a period of several years at a single site, sometimes discontinuously, indicating (latent) survival of the pathogen at those sites.


Statin treatment and diabetes affect myeloperoxidase activity in maintenance hemodialysis patients.
Myeloperoxidase (MPO), which is secreted during activation of neutrophils, may serve as one mechanistic link among persistent inflammation, oxidative stress, and cardiovascular disease. This study related MPO activity to inflammatory and oxidative stress biomarkers, comorbidity, and ongoing medication in prevalent hemodialysis (HD) patients. In a cross-sectional evaluation of 115 prevalent (vintage 25 mo) HD patients (62 men; 63 +/- 1 yr), data on comorbidity (Davies score), diabetes, medication (statins and antihypertensive drugs), nutritional status (subjective global assessment), blood lipids (cholesterol, HDL cholesterol, and triglycerides), inflammatory biomarkers (serum albumin, C-reactive protein, TNF-alpha, and IL-6), oxidative stress biomarkers (pentosidine, 8-hydroxydeoxyguanosine, and MPO activity) were recorded. Patients with MPO activity greater than the median had significantly (P < 0.05) lower serum albumin levels (33.2 +/- 0.7 versus 35.0 +/- 0.5 g/L), higher 8-hydroxydeoxyguanosine levels (1.26 +/- 0.08 versus 1.05 +/- 0.06 ng/ml), and a lower prevalence of statin treatment (18 versus 36%). Therefore, the median MPO activity was significantly (P < 0.05) lower (17.7 versus 26.6 deltaOD630/min per mg protein) in the subgroup of 31 HD patients with ongoing statin treatment. In a multiple regression model, correction for the impact of age, gender, vintage, serum cholesterol, serum albumin, comorbidity, diabetes, and statin use, only diabetes (P < 0.01) and statin use (P < 0.01) were significantly associated to MPO activity. Fourteen patients who had diabetes and were receiving statin treatment had markedly (P = 0.001) lower median (19.9 versus 41.2 deltaOD630/min per mg protein) MPO activity compared with 18 who had diabetes and were not taking statins. This cross-sectional study suggests that both diabetes and statin treatment affect MPO activity in prevalent HD patients.


Stress-related genes define essential steps in the response of maize seedlings to smoke-water.
Smoke from burning vegetation is widely recognised as a germination cue for seed germination and recent reports suggest that smoke treatments can improve seedling vigour also. We investigated the effect of smoke-water on seedling vigour and changes of the global transcriptome in the early post-germination phase in maize. Application of smoke-water improved the germination characteristics and seedling vigour. The transcriptional response of embryos and emerging radicles 24 and 48 h after the onset of smoke treatment was investigated. The microarray study revealed a number of smoke-responsive genes amongst which stress- and abscisic acid (ABA)-related genes were over-represented. The global promoter analysis of the smoke-responsive genes revealed a tight correlation with the results obtained from Gene Ontology annotations. This concerted over-expression shows that smoke treatment induces stress and ABA-related responses in the early post-germination phase which leads to better adaptation to environmental stress factors occurring during germination, eventually resulting in greater seedling vigour.


Mindful Parenting and Parents' Emotion Expression: Effects on Adolescent Risk Behaviors.
Mindful parenting is associated with greater adjustment and fewer behavior problems in children and adolescents. However, the mechanisms by which mindful parenting functions to mitigate risk in adolescence is not well understood. This study investigated parent emotional expression as a potential mechanism in the relationship between low mindful parenting and adolescent risk behaviors. A sample of 157 12-14 year old adolescents (49% female) and their primary caregivers (99% female) participated in an emotionally-arousing conflict interaction. Parents reported on their mindful parenting practices, and parents' emotion expressions during the conflict interaction were coded including negative emotion, positive emotion, and shared parent-youth positive emotion. Adolescent substance use and sex behaviors were assessed through self-report, interview, and physical toxicology screens. Results indicated that mindful parenting was associated with less parental negative emotion and greater shared positive emotion during the parent-adolescent conflict. Further, results revealed a significant indirect effect of mindful parenting on youth's substance use through shared parent-adolescent positive emotion. Findings highlight the relevance of emotional functioning in the context of stressful parenting situations in mindful parenting.


Using computerized virtual cases to explore diagnostic error in practicing physicians.
Background Diagnostic errors are a significant cause of patient harm. Cognitive processes often contribute to diagnostic errors but studying and mitigating the effects of these errors is challenging. Computerized virtual patients may provide insight into the diagnostic process without the potential for patient harm, but the feasibility and utility of using such cases in practicing physicians has not been well described. Methods We developed a series of computerized virtual cases depicting common presentations of disease that included contextual factors that could result in diagnostic error. Cases were piloted by practicing physicians in two phases and participant impressions of the case platform and cases were recorded, as was outcome data on physician performance. Results Participants noted significant challenges in using the case platform. Participants specifically struggled with becoming familiar with the platform and adjusting to the non-adaptive and constraining processes of the model. Although participants found the cases to be typical presentations of problems commonly encountered in practice, the correct diagnosis was identified in less than 33% of cases. Conclusions The development of virtual patient cases for use by practicing physicians requires substantial resources and platforms that account for the non-linear and adaptive nature of reasoning in experienced clinicians. Platforms that are without such characteristics may negatively affect diagnostic performance. The novelty of such platforms may also have the potential to increase cognitive load. Nonetheless, virtual cases may have the potential to be a safe and robust means of studying clinical reasoning performance.


Pathologic characteristics of cancers detected in The Prostate Cancer Prevention Trial: implications for prostate cancer detection and chemoprevention.
The Prostate Cancer Prevention Trial (PCPT) showed a risk of prostate cancer at prostate-specific antigen (PSA) <4.0 ng/mL and that prostate cancer risk is reduced by finasteride. A major concern about early detection by PSA and prevention by finasteride is that they may involve biologically inconsequential tumors. We reviewed the pathologic characteristics of prostate biopsies from men in the placebo and finasteride groups of the PCPT. We examined tumor pathology characteristics stratified by level of PSA for men in the placebo group who underwent radical prostatectomy. Seventy-five percent of all cancers and 62% of Gleason score <or=6 cancers in the PCPT met the biopsy criteria for clinically significant tumors. Surrogate measures for tumor volume (number of cores positive, percent cores positive, linear extent, and bilaterality) and risk of perineural invasion were lower in men who received finasteride. The PSA-associated risks of insignificant cancer were 51.7% (PSA, 0-1.0 ng/mL), 33.7% (1.1-2.5 ng/mL), 17.8% (2.6-4.0 ng/mL), and 11.7% (4.1-10 ng/mL). Conversely, the risks of high-grade (Gleason score >or=7) tumors for the same PSA strata were 15.6%, 37.9%, 49.1%, and 52.4%, respectively. These data highlight the dilemma of PSA when used for screening: Lower cutoff levels increase detection of insignificant disease, but cure is more likely, whereas higher cutoff levels make detection of significant cancer more likely, but cure is less likely. Therefore, the effectiveness of finasteride in preventing prostate cancer, including Gleason score <or=6 cancer, with meaningful rates of significant disease in the PCPT suggests that cutoff values for PSA screening should be individualized and that men undergoing screening should be informed of the opportunity to reduce their risk of disease with finasteride.


Agrochemicals in field margins--assessing the impacts of herbicides, insecticides, and fertilizer on the common buttercup (Ranunculus acris).
The effects of herbicide, insecticide, and fertilizer inputs on the common buttercup Ranunculus acris in field margins were studied in an experimental field study. The test design allowed us to investigate the single and combined effects of repeated herbicide, insecticide, and fertilizer applications in successive growing seasons. To assess the effects of the agrochemical applications on R. acris, plant community assessments were carried out and a photodocumentation of the flowering intensity was performed over two years. In addition, the authors conducted a monitoring survey of R. acris in field margins in the proximity of the study site. In the field experiment, R. acris plant density decreased significantly with treatments including fertilizer. The herbicide caused a sublethal effect by reducing flower intensity by 85%. In the long run, both effects will result in a decline of R. acris and lead to shifts in plant communities in field margins. This was confirmed by the monitoring survey, where R. acris could hardly be observed in field margins directly adjacent to cereal fields, whereas in margins next to meadows the species was recorded frequently. Besides the implications for the plants, the sublethal effects may also affect many flower-visiting insects. The results indicate that the current risk assessment for nontarget plants is insufficiently protective for wild plant species in field margins and that consideration of sublethal effects is crucial to preserve biodiversity in agricultural landscapes.


What do older people experiencing loneliness think about primary care or community based interventions to reduce loneliness? A qualitative study in England.
Loneliness in later life is a common problem with poor health outcomes. However, interventions to prevent or ameliorate loneliness have a weak evidence base. The views of older people experiencing or at risk of loneliness in the community are important in identifying features of potential support, but have been little studied. Twenty-eight community dwelling people, aged 65 and over who reported being 'lonely much of the time' or identified as lonely from the de Jong Gierveld six-item loneliness scale in a larger study, participated in in-depth interviews, between June 2013 and May 2014. Views and experiences on seeking support from primary care and community based one-to-one and group based activities, including social and shared interest groups, were explored. Interviews were recorded and transcribed. Thematic analysis was conducted by a multidisciplinary team, including older people. Using two different measures of loneliness enabled a spectrum of loneliness experience to be explored. Two-thirds of the participants were the 'younger old' and all were able to leave their homes independently. Older people with characteristics of loneliness were generally knowledgeable about local social and community resources but, for the majority, community and primary care based services for their loneliness were not considered desirable or helpful at this point in their lives. However, group based activities with a shared interest were thought preferable to one-to-one support (befriending) or groups with a social focus. Descriptions of support as being for loneliness and specific to older people discouraged engagement. Older people experiencing or at risk of loneliness did not consider that primary care has a role in alleviating loneliness because it is not an illness. They thought primary care practitioners lack understanding of non-physical problems and that a good relationship was necessary to discuss sensitive issues like loneliness. For many, loneliness was a complex and private matter that they wished to manage without external support.


Function and Evolution of C2H2 Zinc Finger Arrays.
Krüppel-type or C2H2 zinc fingers represent a dominant DNA-binding motif in eukaryotic transcription factor (TF) proteins. In Krüppel-type (KZNF) TFs, KZNF motifs are arranged in arrays of three to as many as 40 tandem units, which cooperate to define the unique DNA recognition properties of the protein. Each finger contains four amino acids located at specific positions, which are brought into direct contact with adjacent nucleotides in the DNA sequence as the KZNF array winds around the major groove of the alpha helix. This arrangement creates an intimate and potentially predictable relationship between the amino acid sequence of KZNF arrays and the nucleotide sequence of target binding sites. The large number of possible combinations and arrangements of modular KZNF motifs, and the increasing lengths of KZNF arrays in vertebrate species, has created huge repertoires of functionally unique TF proteins. The properties of this versatile DNA-binding motif have been exploited independently many times over the course of evolution, through attachment to effector motifs that confer activating, repressing or other activities to the proteins. Once created, some of these novel inventions have expanded in specific evolutionary clades, creating large families of TFs that are lineage- or species-unique. This chapter reviews the properties and their remarkable evolutionary history of eukaryotic KZNF TF proteins, with special focus on large families that dominate the TF landscapes in different metazoan species.


Sparse stochastic processes and discretization of linear inverse problems.
We present a novel statistically-based discretization paradigm and derive a class of maximum a posteriori (MAP) estimators for solving ill-conditioned linear inverse problems. We are guided by the theory of sparse stochastic processes, which specifies continuous-domain signals as solutions of linear stochastic differential equations. Accordingly, we show that the class of admissible priors for the discretized version of the signal is confined to the family of infinitely divisible distributions. Our estimators not only cover the well-studied methods of Tikhonov and l1-type regularizations as particular cases, but also open the door to a broader class of sparsity-promoting regularization schemes that are typically nonconvex. We provide an algorithm that handles the corresponding nonconvex problems and illustrate the use of our formalism by applying it to deconvolution, magnetic resonance imaging, and X-ray tomographic reconstruction problems. Finally, we compare the performance of estimators associated with models of increasing sparsity.


The polymerase chain reaction.
The polymerase chain reaction (PCR) is a novel technique that amplifies specific sequences with remarkable efficiency. Repeated cycles of denaturation, primer annealling and extension carried out with the heat stable enzyme, Taq polymerase, leads to exponential increases in the target DNA sequences. The technique has been widely applied in immunology to define HLA polymorphism, T-cell receptor and immunoglobulin diversity, pathogen detection, quantification of lymphokines and lymphoma or leukaemia detection. In addition, it has been very widely applied in all areas of molecular biology and human genetics. Because of its simplicity and sensitivity, PCR represents a major technical development for molecular biologists and will continue to have a great impact on molecular immunology. This review by John Bell describes the basic PCR technique and some variations on it and briefly explores their uses in molecular immunology.


Implementation of a Progressive Mobilization Program in a Medical-Surgical Intensive Care Unit.
Current literature supports implementation of progressive mobility protocols in intensive care units. Education can help nurses overcome barriers to mobility and increase knowledge about the positive effects of mobility. To evaluate the effect of education for a progressive mobilization program for intensive care nurses on knowledge and performance. A pretest-posttest evaluation was conducted for 41 nurses, and a chart review was performed before and after implementation of the educational intervention to evaluate changes in knowledge and mobilization. Scores after the educational intervention were significantly higher than scores before the intervention (t = 2.02; P < .001). Overall mobilization (P = .04) and dangling (P = .01) increased significantly after the education. No significant increases occurred in ambulating or getting patients up to a chair. Mobilization education was effective and increased nurses' knowledge about the benefits of mobility for critically ill patients. The educational program also affected how nurses performed mobility interventions. Although provision of education had positive effects on patients' mobility, leadership and coaching are still important components in implementing change.


Methanogenic food web in the gut contents of methane-emitting earthworm Eudrilus eugeniae from Brazil.
The anoxic saccharide-rich conditions of the earthworm gut provide an ideal transient habitat for ingested microbes capable of anaerobiosis. It was recently discovered that the earthworm Eudrilus eugeniae from Brazil can emit methane (CH4) and that ingested methanogens might be associated with this emission. The objective of this study was to resolve trophic interactions of bacteria and methanogens in the methanogenic food web in the gut contents of E. eugeniae. RNA-based stable isotope probing of bacterial 16S rRNA as well as mcrA and mrtA (the alpha subunit of methyl-CoM reductase and its isoenzyme, respectively) of methanogens was performed with [(13)C]-glucose as a model saccharide in the gut contents. Concomitant fermentations were augmented by the rapid consumption of glucose, yielding numerous products, including molecular hydrogen (H2), carbon dioxide (CO2), formate, acetate, ethanol, lactate, succinate and propionate. Aeromonadaceae-affiliated facultative aerobes, and obligate anaerobes affiliated to Lachnospiraceae, Veillonellaceae and Ruminococcaceae were associated with the diverse fermentations. Methanogenesis was ongoing during incubations, and (13)C-labeling of CH4 verified that supplemental [(13)C]-glucose derived carbon was dissimilated to CH4. Hydrogenotrophic methanogens affiliated with Methanobacteriaceae and Methanoregulaceae were linked to methanogenesis, and acetogens related to Peptostreptoccocaceae were likewise found to be participants in the methanogenic food web. H2 rather than acetate stimulated methanogenesis in the methanogenic gut content enrichments, and acetogens appeared to dissimilate supplemental H2 to acetate in methanogenic enrichments. These findings provide insight on the processes and associated taxa potentially linked to methanogenesis and the turnover of organic carbon in the alimentary canal of methane-emitting E. eugeniae.


Induction of tyrosine aminotransferase in utero by anti-insulin agents.
The hepatic enzyme tyrosine aminotransferase, normally expressed in very low amounts until shortly after birth, is prematurely induced in foetal rats made diabetic by the administration of streptozotocin in utero. Similarly, the enzyme is precociously induced in foetuses if the circulating insulin concentration is artificially decreased by the administration of anti-insulin serum. These observations support the proposal that the natural decrease in plasma insulin, known to occur at birth, is a major contributor to the postnatal induction of tyrosine aminotransferase.


American Gastroenterological Association-American College of Gastroenterology Clinical Practice Guideline: Pharmacological Management of Chronic Idiopathic Constipation.
Chronic idiopathic constipation (CIC) is a common disorder associated with significant impairment in quality of life. This clinical practice guideline, jointly developed by the American Gastroenterological Association and the American College of Gastroenterology, aims to inform clinicians and patients by providing evidence-based practice recommendations for the pharmacological treatment of CIC in adults. The American Gastroenterological Association and the American College of Gastroenterology formed a multidisciplinary guideline panel that conducted systematic reviews of the following agents: fiber, osmotic laxatives (polyethylene glycol, magnesium oxide, lactulose), stimulant laxatives (bisacodyl, sodium picosulfate, senna), secretagogues (lubiprostone, linaclotide, plecanatide), and serotonin type 4 agonist (prucalopride). The panel prioritized clinical questions and outcomes and used the Grading of Recommendations Assessment, Development, and Evaluation framework to assess the certainty of evidence for each intervention. The Evidence to Decision framework was used to develop clinical recommendations based on the balance between the desirable and undesirable effects, patient values, costs, and health equity considerations. The panel agreed on 10 recommendations for the pharmacological management of CIC in adults. Based on available evidence, the panel made strong recommendations for the use of polyethylene glycol, sodium picosulfate, linaclotide, plecanatide, and prucalopride for CIC in adults. Conditional recommendations were made for the use of fiber, lactulose, senna, magnesium oxide, and lubiprostone. This document provides a comprehensive outline of the various over-the-counter and prescription pharmacological agents available for the treatment of CIC. The guidelines are meant to provide a framework for approaching the management of CIC; clinical providers should engage in shared decision making based on patient preferences as well as medication cost and availability. Limitations and gaps in the evidence are highlighted to help guide future research opportunities and enhance the care of patients with chronic constipation.


Disease-free survival according to the use of postmastectomy radiation therapy after neoadjuvant chemotherapy.
The purpose of the study was to determine predictors of recurrence for patients treated with neoadjuvant chemotherapy (NAC) and mastectomy according to the use of postmastectomy radiation therapy (PMRT). An analysis of 161 clinically staged T1 to T3/N0 to N3 patients treated with NAC and mastectomy with and without PMRT at our institution from 2003 to 2010 was conducted. The Kaplan-Meier product limit method was used to estimate survival and time to recurrence rates and the log-rank test was used to compare groups. A Cox proportional hazard regression analysis was carried out for time to recurrence, radiation therapy, and their interaction in the model. The median follow-up period was 48 months and 18 patients developed a recurrence. The 5-year recurrence rate and overall survival was 16.1% (95% confidence interval [CI], 9.6%-26.3%) and 93.6% (95% CI, 88.2%-97.0%), respectively. Patients who underwent PMRT had a decreased risk of recurrence compared with patients who did not (hazard ratio [HR], 0.25; 95% CI, 0.097-0.661; P < .005). The 5-year disease-free survival (DFS) rate for those who received PMRT was 91.3% (95% CI, 82.8%-95.7%) and 64.8% (95% CI, 37.8%-82.4%) for those who did not (P = .0126). Among all clinicopathologic factors examined, pathologic T stage (ypT) and pathological N stage (ypN) significantly correlated with the risk of recurrence (P < .05). Patients with any pathological nodal disease had an increased risk of recurrence compared with patients who were pathologically node-negative (HR, 7.196; 95% CI, 2.05-25.264; P < .002). Patients treated with NAC and mastectomy, but without PMRT had a higher risk recurrence with increasing ypT and ypN stages. PMRT might increase DFS.


Dissection of oenocytes from adult Drosophila melanogaster.
In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides protection against desiccation and other environmental challenges. Several of these cuticular hydrocarbon (CHC) compounds also function as semiochemical signals, and as such mediate pheromonal communications between members of the same species, or in some instances between different species, and influence behavior. Specialized cells referred to as oenocytes are regarded as the primary site for CHC synthesis. However, relatively little is known regarding the involvement of the oenocytes in the regulation of the biosynthetic, transport, and deposition pathways contributing to CHC output. Given the significant role that CHCs play in several aspects of insect biology, including chemical communication, desiccation resistance, and immunity, it is important to gain a greater understanding of the molecular and genetic regulation of CHC production within these specialized cells. The adult oenocytes of D. melanogaster are located within the abdominal integument, and are metamerically arrayed in ribbon-like clusters radiating along the inner cuticular surface of each abdominal segment. In this video article we demonstrate a dissection technique used for the preparation of oenocytes from adult D. melanogaster. Specifically, we provide a detailed step-by-step demonstration of (1) how to fillet prepare an adult Drosophila abdomen, (2) how to identify the oenocytes and discern them from other tissues, and (3) how to remove intact oenocyte clusters from the abdominal integument. A brief experimental illustration of how this preparation can be used to examine the expression of genes involved in hydrocarbon synthesis is included. The dissected preparation demonstrated herein will allow for the detailed molecular and genetic analysis of oenocyte function in the adult fruit fly.


Quantitative RT-PCR assays show Xist RNA levels are low in mouse female adult tissue, embryos and embryoid bodies.
We have investigated expression of the Xist gene in mouse female adult kidney, embryos and embryonic stem (ES) cells undergoing in vitro differentiation as embryoid bodies. Using the quantitative RT-PCR single nucleotide primer extension (SNuPE) assay, we found that the amount of Xist RNA in adult kidney of three mouse strains was less than approximately 2000 transcripts per cell, with only modest differences between strains carrying different Xce alleles. Female embryos 7.5 days post coitum had the same number of Xist transcripts per cell as isogenic adult tissue. Using quantitative oligonucleotide hybridization assays after RT-PCR, we investigated Xist expression in ES lines heterozygous at the Pgk-1 and Xist loci. We found that, while in most (XX) ES lines Xist RNA levels increased during embryoid body formation, the levels seen were less than 10% those found in adult female kidney. In addition, we found that the allelic ratio of Xist transcripts from reciprocal (XX) ES cell lines differentiating in vitro was identical to that of isogenic 10.5 to 11.5 day female embryos. These latter results suggest that there is no pattern of preferential paternal imprinting during days 1 to 9 of in vitro differentiation of ES cells. However, the influence of the Xce locus on the randomness of X-inactivation in embryos seems to operate also in ES cell lines. Our overall conclusion is that the low levels of Xist RNA in female kidney, embryos and differentiating (XX) ES cells are compatible only with models that do not require Xist RNA to cover the entire inactive X chromosome.


Long-term importance of fundamental motor skills: a 20-year follow-up study.
The purpose of this study was to investigate the potential long-term association of motor skill proficiency at 6 years of age and self-reported physical activity (PA) at age 26. Direct motor performance data were collected in 1991 with a follow-up study occurring in 1996, and then indirect questionnaires (self-report) administered in 2001 and 2011. In 2011, 17 participants who were identified as either having high motor proficiency (HMP) or low motor proficiency (LMP) in 1991 completed a series of 4 questionnaires. Analyses were conducted to determine whether there were differences between groups for motor skill proficiency, PA, or sedentary behavior, and whether these outcomes were related across ages. Motor skill proficiency at age 6 was related to self-reported proficiency at age 16 (r = .77, p = .006), and self-reported proficiency between 16 and 26 years (r = .85, p = .001). Motor skill proficiency at age 6 was positively associated with leisure time PA at age 26 in females and participants in the HMP group. The results may provide preliminary evidence about the importance of how early motor skill proficiency relates to long-term PA. More research with larger sample sizes is needed to investigate the importance of motor skills over time.


Recent progress and future directions in protein-protein docking.
This article gives an overview of recent progress in protein-protein docking and it identifies several directions for future research. Recent results from the CAPRI blind docking experiments show that docking algorithms are steadily improving in both reliability and accuracy. Current docking algorithms employ a range of efficient search and scoring strategies, including e.g. fast Fourier transform correlations, geometric hashing, and Monte Carlo techniques. These approaches can often produce a relatively small list of up to a few thousand orientations, amongst which a near-native binding mode is often observed. However, despite the use of improved scoring functions which typically include models of desolvation, hydrophobicity, and electrostatics, current algorithms still have difficulty in identifying the correct solution from the list of false positives, or decoys. Nonetheless, significant progress is being made through better use of bioinformatics, biochemical, and biophysical information such as e.g. sequence conservation analysis, protein interaction databases, alanine scanning, and NMR residual dipolar coupling restraints to help identify key binding residues. Promising new approaches to incorporate models of protein flexibility during docking are being developed, including the use of molecular dynamics snapshots, rotameric and off-rotamer searches, internal coordinate mechanics, and principal component analysis based techniques. Some investigators now use explicit solvent models in their docking protocols. Many of these approaches can be computationally intensive, although new silicon chip technologies such as programmable graphics processor units are beginning to offer competitive alternatives to conventional high performance computer systems. As cryo-EM techniques improve apace, docking NMR and X-ray protein structures into low resolution EM density maps is helping to bridge the resolution gap between these complementary techniques. The use of symmetry and fragment assembly constraints are also helping to make possible docking-based predictions of large multimeric protein complexes. In the near future, the closer integration of docking algorithms with protein interface prediction software, structural databases, and sequence analysis techniques should help produce better predictions of protein interaction networks and more accurate structural models of the fundamental molecular interactions within the cell.


Human T cell lymphotropic virus infection in Guaymi Indians from Panama.
Preliminary studies found that 9% of Guaymi Indians from Bocas del Toro province have antibody to human T cell lymphotropic virus (HTLV-I/II). The present study enrolled 317 (21% of the population) Guaymi Indians from Changuinola, the capital of Bocas del Toro province and 333 (70% of the population) from Canquintu, an isolated rural village. Demographic information and family relationships were ascertained and subjects were screened for neurologic diseases. Serum specimens were screened by an enzyme-linked immunosorbent assay for HTLV-I/II antibody and positives were confirmed according to U.S. Public Health Service criteria. Twenty-five (8%) Guaymi residing in Changuinola and 7 (2.1%) from Canquintu were confirmed seropositive. In Changuinola, antibody was virtually limited to residents greater than or equal to 15 years of age (24 [16%] of 153) and rates were slightly higher in males than in females; in Canquintu, antibody rates did not increase significantly with age and appeared higher in females than in males. In Changuinola, there was no evidence for household clustering of infection. In contrast, HTLV antibody among Canquintu residents clustered significantly by household. HTLV-associated neurologic disease was not detected in either population. The atypical seroepidemiology observed in both locations might be explained if the virus endemic to the Guaymi differed from HTLV-I previously described in the Caribbean basin and Japan.


[Alternative healing methods around us].
This article gives information on selected alternative medicine methods (AMM) under conditions of the Czech health care system. They can be defined as contemporaneous diagnostic and therapeutic methods which differ in their theoretical fundaments as well as practices from the evidence based medicine. This definition is controversial because these methods are an intrinsically incompatible aggregate of modalities acting as a placebo above all. We can encounter many "therapeutic" and "examination" methods which are practiced namely by healers without formal medical education, and connected closely with esotericism. A general feature of the alternative methods is a void usage of scientific concepts and theories, e.g. the concept of energy.Motivations of patients and doctors are briefly introduced here. However, AMM is also an ethical problem because the doctor should help the patient with maximum efficiency hence excluding the methods with a questionable, small or negligible efficiency. The opinions how to find the way out from this situation are divergent. It could be a refusal or a very reserved attitude to AMM plus maximisation of the placebo effect enhancing the efficiency of a rational therapy. Furthermore, the methods most spread in the Czech Republic are subjected to elementary critics - the homoeopathy, acupuncture, the so-called "bioresonance" instrumental methods, psychic healing, and also some refused methods in cancer treatment. In our opinion medicine disposes of sufficient knowledge and experience to assess the efficiency of AMM and refuse them in causal therapy, eventually. Their use as a placebo should be carefully considered.Key words: alternative medicine, placebo, homeopathy, acupuncture.


Evolution of hepatitis C virus quasispecies in hypervariable region 1 and the putative interferon sensitivity-determining region during interferon therapy and natural infection.
To study hepatitis C virus (HCV) genetic mutation during interferon (IFN) therapy, the temporal changes in HCV quasispecies heterogeneity were compared before and after treatment for nine patients infected with HCV genotype 1, including four nonresponders, four responders who relapsed after therapy, and one responder who experienced a breakthrough of viremia during therapy. Nine untreated patients with an average time between specimens of 8.4 years served as controls. Sequences from the second envelope glycoprotein gene hypervariable region 1 (HVR1) and the putative IFN sensitivity-determining region (ISDR) of the nonstructural NS5A gene were analyzed by heteroduplex mobility assays and nucleotide sequencing. A strong positive correlation was found between the percent change in a heteroduplex mobility ratio (HMR) and percent change in nucleotide sequence (r = 0.941, P < 0.001). The rate of fixation of mutations in the HVR1 was significantly higher for IFN-treated patients than for controls (6.97 versus 1.31% change in HMR/year; P = 0.02). Similarly, a higher rate of fixation of mutations was observed in the ISDR for IFN-treated patients than for untreated controls, although the result was not significant (1.45 versus 0.15 amino acid changes/year; P = 0.12). On an individual patient basis, IFN therapy was associated with measurable HVR1 and ISDR mutation in nine of nine (100%) and two of nine (22.2%) patients, respectively. Evolution to IFN-resistant ISDR sequences was observed in only one of nine IFN-treated patients. These data suggest that IFN therapy frequently exerts pressure on the HCV envelope region, while pressure on the ISDR was evident in only a subset of patients. Thus, the selection pressures evoked on HCV genotype 1 quasispecies during IFN therapy appear to differ among different patients.


Health economic aspects of late preterm and early term birth.
Despite an increasing body of knowledge on the adverse clinical sequelae associated with late preterm birth and early term birth, little is known about their economic consequences or the cost-effectiveness of interventions aimed at their prevention or alleviation of their effects. This review assesses the health economic evidence surrounding late preterm and early term birth. Evidence is gathered on hospital resource use associated with late preterm and early term birth, economic costs associated with late preterm and early term birth, and economic evaluations of prevention and treatment strategies. The article highlights the limited perspective and time horizon of most studies of economic costs in this area; the limited evidence surrounding health economic aspects of early term birth; the gaps in current knowledge; and it discusses directions for future research in this area, including the need for validated tools for measuring preference-based health-related quality-of-life outcomes in infants that will aid cost-effectiveness-based decision-making.


Bioremediation of Cr(VI) and immobilization as Cr(III) by Ochrobactrum anthropi.
Bioremediation of Cr(VI) through reduction relies on the notion that the produced Cr(III) may be precipitated or efficiently immobilized. However, recent reports suggest that soluble organo-Cr(III) complexes are present in various chromate-reducing bacterial systems. This work was designed to explore the factors that affect the immobilization of Cr(III) in the Ochrobactrum anthropi system. X-ray absorption fine structure analysis on the cell debris clearly verified that coordination of Cr(III) occurs on the surfaces via the chelating coordination with carboxyl- and amido-functional groups. However, competitive coordination experiments of Cr(III) revealed that the small molecules such as amino acids and their derivatives or multicarboxyl compounds hold stronger coordination ability with Cr(III) than with cell debris. We speculate that it is the preferential coordination of Cr(III) to the soluble organic molecules in the bacterial culture medium that inhibits effective immobilization of Cr(III) on the cells. On the basis of this understanding, a strategy with two-step control of the medium was proposed, and this achieved successful immobilization of Cr(VI) as Cr(III) by O. anthropi and Planococcus citreus in 5-50 L pilot-scale experiments.


[Complement-mediated solubilization of immune complexes in immune complex nephritis].
In order to examine the ability of complement to solubilize the immune complexes (IC) in vivo, we made acute or chronic serum sickness nephritis in 10 rabbits or 12 rats with injection of BSA as antigen (Ag). Animals were divided into two groups. The 500 mu or 50 mu/week of cobra venom factor (CVF) for rabbits or rats was administered intraperitoneally in one group of animals to reduce the serum complement levels and complement-mediated solubilization of immune complex. On the other hand, the 2 ml or 0.2 ml/week of turpentine oil (TO) was injected intramuscularly in another group of animals to elevate the serum complement levels. The renal biopsy was performed with weekly intervals in each animals, and the specimens were stained for antigen (BSA), antibody (IgG) and C3 by immunofluorescence techniques. The result in this experiments showed the significant difference between TO-, and CVF-treated animals in the degree of disappearance of IC in the kidney lesions. The undetectable amount of IC (especially BSA) was remained in the renal tissue of TO-treated animals. This fact indicated that complement might act to solubilize the IC deposited in the tissue in vivo also.


Cutaneous neoplastic lesions of equids in the central United States and Canada: 3,351 biopsy specimens from 3,272 equids (2000-2010).
To identify epidemiological trends in cutaneous neoplasms affecting equids in central North America and compare them with previously reported trends. Retrospective case series. 3,351 cutaneous biopsy specimens from 3,272 equids with a neoplastic diagnosis. Diagnostic reports from 2 diagnostic laboratories (Colorado State University and Prairie Diagnostic Services Inc) were reviewed for frequency of specific lesions and epidemiological trends. Variables included in analyses (if known) were age, sex, breed, geographic location, date of diagnosis, location of neoplasm on the body, and presence or absence of ulceration. Sarcoid, squamous cell carcinoma (SCC), and melanoma were the 3 most common tumors diagnosed. Tumors associated with UV radiation (SCC, SCC in situ, hemangioma, hemangiosarcoma) were 2.3 (95% confidence interval [CI], 1.8 to 3.0) times as common in biopsy specimens received by Colorado State University than in specimens received by Prairie Diagnostic Services Inc. Appaloosa horses and American Paint horses, respectively, were 7.2 (95% CI, 5.6 to 9.2) and 4.4 (95% CI, 3.6 to 5.4) times as likely as other breeds to have tumors associated with UV radiation. Thoroughbreds were predisposed to cutaneous lymphoma, whereas Arabians were more likely to have melanomas. Draft and pony breeds were 3.1 (95% CI, 1.9 to 5.1) times as likely as other breeds to have benign soft tissue tumors. Morgans and pony breeds more commonly had basal cell tumors. Tumors in the perianal region were significantly more likely to be SCC or melanoma while tumors on the limbs were more likely to be giant cell tumor of soft parts. Signalment, anatomic location of the mass, and geographic location of the horse can be used to help equine practitioners formulate differential diagnoses for cutaneous masses. Further research is necessary to identify the biological basis for the development of many equine cutaneous neoplasms.


Dietary patterns and breast cancer risk in the California Teachers Study cohort.
Evidence that diet is associated with breast cancer risk is inconsistent. Most studies have examined risks associated with specific foods and nutrients, rather than measures of overall diet. This study aimed to evaluate dietary patterns and their relation to breast cancer risk in a large cohort of women. Data from 91,779 women in the California Teachers Study cohort were analyzed, including data from 4140 women with a diagnosis of invasive breast cancer made between 1995 and 2009. Five predominant dietary patterns were identified by using principal components factor analysis: a plant-based diet, high in fruit and vegetables; a high-protein, high-fat diet, high in meats, eggs, fried foods, and high-fat condiments; a high-carbohydrate diet, high in convenience foods, pasta, and bread products; an ethnic diet, high in legumes, soy-based foods, rice, and dark-green leafy vegetables; and a salad and wine diet, high in lettuce, fish, wine, low-fat salad dressing, and coffee and tea. The plant-based pattern was associated with a reduction in breast cancer risk (RR: 0.85; 95% CI: 0.76, 0.95 for the highest compared with the lowest consumption quintile; P-trend = 0.003); risk reduction was greater for estrogen receptor-negative progesterone receptor-negative (ER-PR-) tumors (RR: 0.66; 95% CI: 0.48, 0.91; P-trend = 0.03). The salad and wine pattern was associated with an increased risk of estrogen receptor-positive progesterone receptor-positive tumors (RR: 1.29; 95% CI: 1.12, 1.49); this effect was only slightly attenuated after adjustment for alcohol consumption. The finding that greater consumption of a plant-based dietary pattern is associated with a reduced breast cancer risk, particularly for ER-PR- tumors, offers a potential avenue for prevention.


Efficacy of sodium hyaluronate and carboxymethylcellulose in treating mild to moderate dry eye disease.
We compared the efficacy and safety of sodium hyaluronate (SH) and carboxymethylcellulose (CMC) in treating mild to moderate dry eye. Sixty-seven patients with mild to moderate dry eye were enrolled in this prospective, randomized, blinded study. They were treated 6 times a day with preservative-free unit dose formula eyedrops containing 0.1% SH or 0.5% CMC for 8 weeks. Corneal and conjunctival staining with fluorescein, tear film breakup time, subjective symptoms, and adverse reactions were assessed at baseline, 4 weeks, and 8 weeks after treatment initiation. Thirty-two patients were randomly assigned to the SH group and 33 were randomly assigned to the CMC group. Both the SH and CMC groups showed statistically significant improvements in corneal and conjunctival staining sum scores, tear film breakup time, and dry eye symptom score at 4 and 8 weeks after treatment initiation. However, there were no statistically significant differences in any of the indices between the 2 treatment groups. There were no significant adverse reactions observed during follow-up. The efficacies of SH and CMC were equivalent in treating mild to moderate dry eye. SH and CMC preservative-free artificial tear formulations appropriately manage dry eye sign and symptoms and show safety and efficacy when frequently administered in a unit dose formula.


Liquid crystal modulator with ultra-wide dynamic range and adjustable driving voltage.
We demonstrated a reflective-type liquid crystal (LC) intensity modulator in 1550 nm telecomm band. An effective way to compensate the residual phase of a LC cell is proposed. With the adjustment of a true zero-order quarter wave plate and enhanced by total internal reflection induced birefringence, over 53 dB dynamic range was achieved, which is much desired for some high-end optical communication, infrared scene projection applications. In addition, the driving voltages were decreased and adjustable. Mechanical and spectral tolerance measurements show that our LC modulator is quite stable. Further applications of our experimental setup were discussed including bio-sensors and high speed modulators.


D-Limonene: safety and clinical applications.
D-limonene is one of the most common terpenes in nature. It is a major constituent in several citrus oils (orange, lemon, mandarin, lime, and grapefruit). D-limonene is listed in the Code of Federal Regulations as generally recognized as safe (GRAS) for a flavoring agent and can be found in common food items such as fruit juices, soft drinks, baked goods, ice cream, and pudding. D-limonene is considered to have fairly low toxicity. It has been tested for carcinogenicity in mice and rats. Although initial results showed d-limonene increased the incidence of renal tubular tumors in male rats, female rats and mice in both genders showed no evidence of any tumor. Subsequent studies have determined how these tumors occur and established that d-limonene does not pose a mutagenic, carcinogenic, or nephrotoxic risk to humans. In humans, d-limonene has demonstrated low toxicity after single and repeated dosing for up to one year. Being a solvent of cholesterol, d-limonene has been used clinically to dissolve cholesterol-containing gallstones. Because of its gastric acid neutralizing effect and its support of normal peristalsis, it has also been used for relief of heartburn and gastroesophageal reflux (GERD). D-limonene has well-established chemopreventive activity against many types of cancer. Evidence from a phase I clinical trial demonstrated a partial response in a patient with breast cancer and stable disease for more than six months in three patients with colorectal cancer.


Genital lesions in experimental chronic Trypanosoma brucei infection in rams.
Rams inoculated intravenously with Trypanosoma brucei developed scrotal oedema and a nonsuppurative granulomatous periorchitis resulting in testicular degeneration, atrophy, calcification and sclerosis. The tunica vaginalis was the seat of an intense granulomatous inflammation and focal necrosis, which extended to the epididymis and spermatic cord. Lesions in the seminal vesicles were suggestive of diminished testosterone production even though Leydig cells were discernible in the sclerotic testicular intertubular tissue. In areas where there was inflammation or focal necrosis, there was also extravascular localisation of trypanosomes. It is suggested that genital lesions in human sleeping sickness may have a similar pathogenetic mechanism involving tissue localisation of the trypanosomes and associated inflammatory and degenerative changes.


Lower socioeconomic status is associated with worse outcomes after both listing and transplanting children with heart failure.
The relationship between SES and outcomes surrounding pediatric cardiac transplantation is complex and influenced by recipient race. Broad-based studies of SES have not been performed. A retrospective review of all 5125 primary pediatric heart transplants performed in the United States between 2000 and 2011. Patients were stratified by SES based on zip code of residence and U.S. census data (low SES: 1637; mid-SES: 2253; high SES: 1235). Survival following listing and transplantation was compared across strata. Risk-adjusted long-term mortality on the waitlist was higher among low SES patients (hazard 1.32, CI 1.07-1.63). The relationship between SES and outcomes varied by race. Early risk-adjusted post-transplant outcomes were worst among high SES patients (10.8% vs. low SES: 8.9%, p < 0.05). The incidence of non-compliance was higher among low SES patients (p < 0.0001). Long-term risk-adjusted patient survival was poorer among low (hazard 1.41, CI 1.10-1.80) and mid-SES (1.29, 1.04-1.59) groups. Low SES is associated with worse outcomes on both the waitlist and late following transplantation. Higher SES patients had more complex transplants with higher early mortality. Further research should be directed at identifying and addressing underlying causal factors for these disparities.


Intratumoral peptide injection enhances tumor cell antigenicity recognized by cytotoxic T lymphocytes: a potential option for improvement in antigen-specific cancer immunotherapy.
Antigen-specific cancer immunotherapy is a promising strategy for improving cancer treatment. Recently, many tumor-associated antigens and their epitopes recognized by cytotoxic T lymphocytes (CTLs) have been identified. However, the density of endogenously presented antigen-derived peptides on tumor cells is generally sparse, resulting in the inability of antigen-specific CTLs to work effectively. We hypothesize that increasing the density of an antigen-derived peptide would enhance antigen-specific cancer immunotherapy. Here, we demonstrated that intratumoral peptide injection leads to additional peptide loading onto major histocompatibility complex class I molecules of tumor cells, enhancing tumor cell recognition by antigen-specific CTLs. In in vitro studies, human leukocyte antigen (HLA)-A*02:01-restricted glypican-3144-152 (FVGEFFTDV) and cytomegalovirus495-503 (NLVPMVATV) peptide-specific CTLs showed strong activity against all peptide-pulsed cell lines, regardless of whether the tumor cells expressed the antigen. In in vivo studies using immunodeficient mice, glypican-3144-152 and cytomegalovirus495-503 peptides injected into a solid mass were loaded onto HLA class I molecules of tumor cells. In a peptide vaccine model and an adoptive cell transfer model using C57BL/6 mice, intratumoral injection of ovalbumin257-264 peptide (SIINFEKL) was effective for tumor growth inhibition and survival against ovalbumin-negative tumors without adverse reactions. Moreover, we demonstrated an antigen-spreading effect that occurred after intratumoral peptide injection. Intratumoral peptide injection enhances tumor cell antigenicity and may be a useful option for improvement in antigen-specific cancer immunotherapy against solid tumors.


Topographic and magnetic-sensitive scanning tunneling microscope study of magnetite.
The topographic and magnetic surface structure of a natural single crystal of magnetite (Fe(3)0(4)), a common mineral, has been studied from the submicrometer scale down to the atomic scale with a scanning tunneling microscope having nonmagnetic tungsten as well as ferromagnetic iron probe tips. Several different (001) crystal planes were imaged to atomic resolution with both kinds of tips. A selective imaging of the octahedrally coordinated Fe B-sites in the Fe-O planes, and even a selective imaging of the different magnetic ions Fe(2+) and Fe(3+), has been achieved, demonstrating for the first time that magnetic imaging can be realized at the atomic level.


"Fulfilling the chief of his duties as a physician": Harvey Cushing, selective dorsal rhizotomy and elective spine surgery for quality of life.
At the beginning of the 20th century, the development of safer anesthesia, antiseptic techniques, and meticulous surgical dissection led to a substantial decrease in operative risk. In turn, the scope of surgery expanded to include elective procedures performed with the intention of improving the quality of life of patients. Between 1908 and 1912, Harvey Cushing performed 3 dorsal rhizotomies to improve the quality of life of 3 patients with debilitating neuralgia: a 54-year-old man with "lightning" radicular pain from tabes dorsalis, a 12-year-old boy cutaneous hyperesthesia and spasticity in his hemiplegic arm, and a 61-year-old man with postamputation neuropathic pain. Symptomatic improvement was seen postoperatively in the first 2 cases, although the third patient continued to have severe pain. Cushing also removed a prominent spinous process from each of 2 patients with debilitating headaches; both patients, however, experienced only minimal postoperative improvement. These cases, which have not been previously published, highlight Cushing's views on the role of surgery and illustrate the broader movement that occurred in surgery at the time, whereby elective procedures for quality of life became performed and accepted.


Phagosome-Bacteria Interactions from the Bottom Up.
When attempting to propagate infections, bacterial pathogens encounter phagocytes that encase them in vacuoles called phagosomes. Within phagosomes, bacteria are bombarded with a plethora of stresses that often lead to their demise. However, pathogens have evolved numerous strategies to counter those host defenses and facilitate survival. Given the importance of phagosome-bacteria interactions to infection outcomes, they represent a collection of targets that are of interest for next-generation antibacterials. To facilitate such therapies, different approaches can be employed to increase understanding of phagosome-bacteria interactions, and these can be classified broadly as top down (starting from intact systems and breaking down the importance of different parts) or bottom up (developing a knowledge base on simplified systems and progressively increasing complexity). Here we review knowledge of phagosomal compositions and bacterial survival tactics useful for bottom-up approaches, which are particularly relevant for the application of reaction engineering to quantify and predict the time evolution of biochemical species in these death-dealing vacuoles. Further, we highlight how understanding in this area can be built up through the combination of immunology, microbiology, and engineering.


The genealogy and genetic viability of reintroduced Yellowstone grey wolves.
The recovery of the grey wolf in Yellowstone National Park is an outstanding example of a successful reintroduction. A general question concerning reintroduction is the degree to which genetic variation has been preserved and the specific behavioural mechanisms that enhance the preservation of genetic diversity and reduce inbreeding. We have analysed 200 Yellowstone wolves, including all 31 founders, for variation in 26 microsatellite loci over the 10-year reintroduction period (1995-2004). The population maintained high levels of variation (1995 H(0) = 0.69; 2004 H(0) = 0.73) with low levels of inbreeding (1995 F(IS) = -0.063; 2004 F(IS) = -0.051) and throughout, the population expanded rapidly (N(1995) = 21; N(2004) = 169). Pedigree-based effective population size ratios did not vary appreciably over the duration of population expansion (1995 N(e)/N(g) = 0.29; 2000 N(e)/N(g) = 0.26; 2004 N(e)/N(g) = 0.33). We estimated kinship and found only two of 30 natural breeding pairs showed evidence of being related (average r = -0.026, SE = 0.03). We reconstructed the genealogy of 200 wolves based on genetic and field data and discovered that they avoid inbreeding through a wide variety of behavioural mechanisms including absolute avoidance of breeding with related pack members, male-biased dispersal to packs where they breed with nonrelatives, and female-biased subordinate breeding. We documented a greater diversity of such population assembly patterns in Yellowstone than previously observed in any other natural wolf population. Inbreeding avoidance is nearly absolute despite the high probability of within-pack inbreeding opportunities and extensive interpack kinship ties between adjacent packs. Simulations showed that the Yellowstone population has levels of genetic variation similar to that of a population managed for high variation and low inbreeding, and greater than that expected for random breeding within packs or across the entire breeding pool. Although short-term losses in variation seem minimal, future projections of the population at carrying capacity suggest significant inbreeding depression will occur without connectivity and migratory exchange with other populations.


The relationship between alcohol and cognitive functioning following traumatic brain injury.
The present study aimed to examine the association between frequency and quantity of alcohol consumption and cognitive functioning following traumatic brain injury (TBI). Sixty moderately to severely injured individuals had completed the Alcohol Use Disorders Identification Test (AUDIT) to measure preinjury alcohol use soon after injury and were recruited and assessed with AUDIT and Time Line Follow-Back (TLFB), as a measure of frequency and quantity of alcohol consumption at 6-9 months post injury. Fifty participants completed both AUDIT and TLFB at a follow-up assessment at 12-15 months post injury. Measures of processing speed/attention, memory, and executive function were also administered. Regression analyses were used to examine the relationships between alcohol use and cognition at the two occasions of measurement. Harmful or hazardous alcohol use pre injury was associated with poorer memory performance on the California Verbal Learning Test (CVLT-II) and slower processing speed on Symbol Digit Modalities Test on average across postinjury measurement occasions, but not with executive functioning, measured by the Modified Six Elements Test (MSET) at 6-9 months post injury. On the other hand, executive functioning on MSET 6-9 months post injury was significantly poorer in participants who were consuming any alcohol at all in the month prior to follow-up assessment. The current study provides evidence showing that pre- and postinjury alcohol use is negatively associated with different aspects of cognitive functioning following TBI. In addition to providing some support for the provision of advice to abstain from alcohol after injury, these findings suggest that interventions to reduce postinjury alcohol use may be useful.


The extreme floods in the Ebro River basin since 1600 CE.
Reliable and complete knowledge of the historical floods is necessary for understanding the extreme hydrological dynamics of the rivers, their natural variability and anthropic changes. In this work we reconstruct the most important floods of the Ebro basin during the last 400 years in different areas of the basin. The analysis is based on four different areas: the Ebro River at Zaragoza, the Cinca River at Fraga, the Segre River at Lleida, and the Ebro River near its mouth at Tortosa. Based on a documentary research, we have first obtained relevant information about the initial conditions (rainfall duration and distribution, snow cover influence) and the maximum flood heights that allow to reconstruct the maximum peak flows by using hydraulic models and to calculate the subbasins contributions. The results show four main types of extreme floods: a) those affecting simultaneously all the subbasins with the highest peak discharges (Ebro at Tortosa in 1787: 0.15 m3 s-1 km-2); b) those originated at the western basin, upstream from Zaragoza, with an Atlantic origin, presenting moderate maximum peak flows, caused by persistent winter rainfall and where snowmelt significantly contributes to the flood; c) those originating at the central Pyrenean subbasins, with Mediterranean origin, occurring, with high peak discharges. These mainly occur during autumn as a consequence of rainfalls of different duration (between 3 days and 1 month), and without significant snow thawing and d) finally, less frequent but very intense flash floods events centered in the Lower Ebro area with low peak flows. In terms of frequency, two different periods can be distinguished: from 1600 until 1850, the frequency of events is low; since 1850 the frequency of events is clearly higher, due to an increase of the climatic variability during last stages of the Little Ice Age. From the 1960's reservoirs construction modifies discharges regime.


Abdominal attacks and treatment in hereditary angioedema with C1-inhibitor deficiency.
Hereditary angioedema (HAE) is characterized by unpredictable attacks of debilitating subcutaneous and mucosal edema. Gastrointestinal attacks are painful, of sudden onset and often mistaken for acute abdomen leading to unnecessary surgery. The purpose of this study was to analyze symptom presentation of gastrointestinal angioedema in pediatric and adult HAE patients. Information collected during the clinical development of ecallantide for treatment of acute HAE attacks included affected anatomic location, accompanying symptoms, medical history, and pain assessments. Efficacy endpoints included Treatment Outcome Score (TOS, maximum score = 100; minimally important difference = 30), a point-in-time measure of treatment response, and time to treatment response. Forty-nine percent of 521 HAE attacks only involved abdominal symptoms. The most commonly reported abdominal symptoms were distension (77%), cramping (73%) and nausea (67%). The most common pain descriptors were tender, tiring-exhausting, aching, cramping and sickening. White blood cell counts were elevated (>10 × 10(9)/L) in 23% of attacks (mean ± SD: 15.1 ± 11.27 × 10(9)/L). A high proportion of patients reported a history of abdominal surgery, including appendectomy (23%), cholecystectomy (16.4%), and hysterectomy (8.2%). Mean TOS at 4 hours post ecallantide was 77 ± 33 versus 29 ± 65 for placebo. Median time to significant symptom resolution was 165 minutes (95% CI 136, 167) for ecallantide versus >4 hours (95% CI 161, >4 hours) for placebo. Anaphylactic reactions occurred in 6 of the 149 treated patients. HAE should be considered in the differential diagnosis of patients with recurrent discrete episodes of severe, unexplained crampy abdominal pain associated with nausea. The data used in the analysis were gathered across multiple clinical trials conducted during the clinical development program for ecallantide. All of the studies were conducted using Good Clinical Practices (GCP) and in accordance with the ethical principles that have their origins in the Declaration of Helsinki. Each site that participated in the clinical trials obtained the appropriate IRB or Ethics Committee approval prior to enrolling any patients. All patients provided written informed consent prior to undergoing any study-related procedures. Pediatric patients provided written assent and their parents or guardians gave written informed consent.The following trials have been registered at http://www.clinicaltrials.gov: EDEMA2 (identifier NCT01826916); EDEMA3 (identifier NCT00262080); EDEMA4 (identifier NCT00457015); and DX-88/19 (identifier NCT00456508).


Cephalometric evaluation of nongrowing females with skeletal and dental Class II, division 1 malocclusion.
Studies evaluating maxillary and mandibular skeletal and dental positions and the vertical components of Class II patients have reported conflicting results. In addition, no common results have been found regarding cranial base configurations. However, few studies have evaluated nongrowing subjects. The aim of this study was to establish whether patients with skeletal and dental Class II division 1 malocclusion have specific craniofacial features. For this, 40 nongrowing females were evaluated cephalometrically. Wide variations were observed for almost all measurements of Class II division 1 patients. However, a posteriorly positioned and rotated mandible, protrusive mandibular incisors, and an increased cranial base angle were all mean characteristics of Class II division 1 patients.


Differential pathogenesis of respiratory syncytial virus clinical isolates in BALB/c mice.
Airway mucus is a hallmark of respiratory syncytial virus (RSV) lower respiratory tract illness. Laboratory RSV strains differentially induce airway mucus production in mice. Here, we tested the hypothesis that RSV strains differ in pathogenesis by screening six low-passage RSV clinical isolates for mucogenicity and virulence in BALB/cJ mice. The RSV clinical isolates induced variable disease severity, lung interleukin-13 (IL-13) levels, and gob-5 levels in BALB/cJ mice. We chose two of these clinical isolates for further study. Infection of BALB/cJ mice with RSV A2001/2-20 (2-20) resulted in greater disease severity, higher lung IL-13 levels, and higher lung gob-5 levels than infection with RSV strains A2, line 19, Long, and A2001/3-12 (3-12). Like the line 19 RSV strain, the 2-20 clinical isolate induced airway mucin expression in BALB/cJ mice. The 2-20 and 3-12 RSV clinical isolates had higher lung viral loads than laboratory RSV strains at 1 day postinfection (p.i.). This increased viral load correlated with higher viral antigen levels in the bronchiolar epithelium and greater histopathologic changes at 1 day p.i. The A2 RSV strain had the highest peak viral load at day 4 p.i. RSV 2-20 infection caused epithelial desquamation, bronchiolitis, airway hyperresponsiveness, and increased breathing effort in BALB/cJ mice. We found that RSV clinical isolates induce variable pathogenesis in mice, and we established a mouse model of clinical isolate strain-dependent RSV pathogenesis that recapitulates key features of RSV disease.


A novel bioactivity of andrographolide from Andrographis paniculata on cerebral ischemia/reperfusion-induced brain injury through induction of cerebral endothelial cell apoptosis.
Andrographolide, extracted from the leaves of Andrographis paniculata (Burm. f.) Nees (Acanthaceae), is a labdane diterpene lactone. It is widely reported to possess anti-inflammatory and antitumorigenic activities. Cerebral endothelial cells (CECs) play a crucial role in supporting the integrity and the function of the blood-brain barrier (BBB). However, no data are available concerning the effects of andrographolide in CECs. The aim of this study was to examine the detailed mechanisms of andrographolide on CECs. This study investigated a novel bioactivity of andrographolide on cerebral ischemia/reperfusion-induced brain injury. CECs were treated with andrographolide (20-100 µΜ) for the indicated times (0-24 h). After the reactions, cell survival rate and cytotoxicity were tested by the MTT assay and the lactate dehydrogenase (LDH) test, respectively. Western blotting was used to detect caspase-3 expression. In addition, analysis of cell cycle and apoptosis using PI staining and annexin V-FITC/PI labeling, respectively, was performed by flow cytometry. We also investigated the effect of andrographolide on middle cerebral artery occlusion (MCAO)/reperfusion-induced brain injury in a rat model. In the present study, we found that andrographolide (50-100 µΜ) markedly inhibited CEC growth according to an MTT assay and caused CEC damage according to a LDH test. Our data also revealed that andrographolide (50 µM) induced CEC apoptosis and caspase-3 activation as respectively detected by PI/annexin-V double staining and western blotting. Moreover, andrographolide arrested the CEC cell cycle at the G0/G1 phase by PI staining. In addition, andrographolide (5 mg/kg) caused deterioration of MCAO/reperfusion-induced brain injury in a rat model. These data suggest that andrographolide may disrupt BBB integrity, thereby deteriorating MCAO/reperfusion-induced brain injury, which are, in part, associated with its capacity to arrest cell-cycle and induce CEC apoptosis.


Aromaticity of metallabenzenes and related compounds.
The concept of aromaticity was initially introduced in chemistry to account for the stability, reactivity, molecular structures, and other properties of many unsaturated organic compounds. Despite that, it has been extended to other species with mobile electrons including saturated systems, transition structures, and even inorganic molecules. In this review, we focus on the aromaticity of a particular family of organometallic compounds known as metallabenzenes, which are characterized by the formal replacement of a CH group in benzene by an isolobal transition metal fragment. In addition, aromaticity of related compounds such as heterometallabenzenes is considered as well. To this end, we shall describe herein the insight gained by the available experimental data as well as by the application of the state-of-the-art computational methods developed as descriptors for aromaticity together with a critical evaluation of their performance to quantitatively estimate the strength of aromaticity of these systems.


Effect of Peroral Endoscopic Myotomy vs Pneumatic Dilation on Symptom Severity and Treatment Outcomes Among Treatment-Naive Patients With Achalasia: A Randomized Clinical Trial.
Case series suggest favorable results of peroral endoscopic myotomy (POEM) for treatment of patients with achalasia. Data comparing POEM with pneumatic dilation, the standard treatment for patients with achalasia, are lacking. To compare the effects of POEM vs pneumatic dilation as initial treatment of treatment-naive patients with achalasia. This randomized multicenter clinical trial was conducted at 6 hospitals in the Netherlands, Germany, Italy, Hong Kong, and the United States. Adult patients with newly diagnosed achalasia and an Eckardt score greater than 3 who had not undergone previous treatment were included. The study was conducted between September 2012 and July 2015, the duration of follow-up was 2 years after the initial treatment, and the final date of follow-up was November 22, 2017. Randomization to receive POEM (n = 67) or pneumatic dilation with a 30-mm and a 35-mm balloon (n = 66), with stratification according to hospital. The primary outcome was treatment success (defined as an Eckardt score ≤3 and the absence of severe complications or re-treatment) at the 2-year follow-up. A total of 14 secondary end points were examined among patients without treatment failure, including integrated relaxation pressure of the lower esophageal sphincter via high-resolution manometry, barium column height on timed barium esophagogram, and presence of reflux esophagitis. Of the 133 randomized patients, 130 (mean age, 48.6 years; 73 [56%] men) underwent treatment (64 in the POEM group and 66 in the pneumatic dilation group) and 126 (95%) completed the study. The primary outcome of treatment success occurred in 58 of 63 patients (92%) in the POEM group vs 34 of 63 (54%) in the pneumatic dilation group, a difference of 38% ([95% CI, 22%-52%]; P < .001). Of the 14 prespecified secondary end points, no significant difference between groups was demonstrated in 10 end points. There was no significant between-group difference in median integrated relaxation pressure (9.9 mm Hg in the POEM group vs 12.6 mm Hg in the pneumatic dilation group; difference, 2.7 mm Hg [95% CI, -2.1 to 7.5]; P = .07) or median barium column height (2.3 cm in the POEM group vs 0 cm in the pneumatic dilation group; difference, 2.3 cm [95% CI, 1.0-3.6]; P = .05). Reflux esophagitis occurred more often in the POEM group than in the pneumatic dilation group (22 of 54 [41%] vs 2 of 29 [7%]; difference, 34% [95% CI, 12%-49%]; P = .002). Two serious adverse events, including 1 perforation, occurred after pneumatic dilation, while no serious adverse events occurred after POEM. Among treatment-naive patients with achalasia, treatment with POEM compared with pneumatic dilation resulted in a significantly higher treatment success rate at 2 years. These findings support consideration of POEM as an initial treatment option for patients with achalasia. Netherlands Trial Register number: NTR3593.