Global analysis of Arabidopsis/downy mildew interactions reveals prevalence of incomplete resistance and rapid evolution of pathogen recognition.
Interactions between Arabidopsis thaliana and its native obligate oomycete pathogen Hyaloperonospora arabidopsidis (Hpa) represent a model system to study evolution of natural variation in a host/pathogen interaction. Both Arabidopsis and Hpa genomes are sequenced and collections of different sub-species are available. We analyzed ∼400 interactions between different Arabidopsis accessions and five strains of Hpa. We examined the pathogen's overall ability to reproduce on a given host, and performed detailed cytological staining to assay for pathogen growth and hypersensitive cell death response in the host. We demonstrate that intermediate levels of resistance are prevalent among Arabidopsis populations and correlate strongly with host developmental stage. In addition to looking at plant responses to challenge by whole pathogen inoculations, we investigated the Arabidopsis resistance attributed to recognition of the individual Hpa effectors, ATR1 and ATR13. Our results suggest that recognition of these effectors is evolutionarily dynamic and does not form a single clade in overall Arabidopsis phylogeny for either effector. Furthermore, we show that the ultimate outcome of the interactions can be modified by the pathogen, despite a defined gene-for-gene resistance in the host. These data indicate that the outcome of disease and disease resistance depends on genome-for-genome interactions between the host and its pathogen, rather than single gene pairs as thought previously.


An Overview of Nanotechnological Approaches for the Diagnosis and Treatment of Allergic Illness.
Allergies are a major health issue. Allergen avoidance, antihistamines, and corticosteroids do not treat the pathology's causes, therefore long-term therapy is essential. Long-term allergen-specific immunotherapy builds immune tolerance to the allergen. Unfortunately, immunotherapies for all allergens are not available, and adverse reactions during therapy, especially in severely allergic persons, remain a worry. In this regard, cell and bio- or nanomaterial-based allergy treatments are promising. This overview covers the most important tactics from these two strategies with examples. Nanotechnology encompasses science, engineering, and technology at 1-100 nm. Due to their one-of-a-kind characteristics, nanomaterials can be used in healthcare. Small molecules' chemical and physical properties are modified by the system's size, shape, content, and function. Toxicity and hypersensitivity reactions need to be evaluated. Regulating the physico-chemical properties of numerous accessible structures would make clinical diagnosis and therapy safer and more successful. Dendrimeric antigens, nanoallergens, and nanoparticles can mimic carrier proteins, boost specific IgE binding, and improve signal detection in allergy diagnosis. In immunotherapy, several allergenic structures like glycodendrimers, liposomes, polymers, and nanoparticles have been used as adjuvants, protectors, or depots for allergens. Nanotechnology has the potential to substantially improve both the diagnosis and treatment of allergies.


Nutrition and the risk of stroke.
Poor nutrition in the first year of a mother's life and undernutrition in utero, infancy, childhood, and adulthood predispose individuals to stroke in later life, but the mechanism of increased stroke risk is unclear. Overnutrition also increases the risk of stroke, probably by accelerating the development of obesity, hypertension, hyperlipidaemia, and diabetes. Reliable evidence suggests that dietary supplementation with antioxidant vitamins, B vitamins, and calcium does not reduce the risk of stroke. Less reliable evidence suggests that stroke can be prevented by diets that are prudent, aligned to the Mediterranean or DASH (Dietary Approaches to Stop Hypertension) diets, low in salt and added sugars, high in potassium, and meet, but do not exceed, energy requirements. Trials in progress are examining the effects of vitamin D and marine omega-3 fatty acid supplementation on incidence of stroke. Future challenges include the need to improve the quality of evidence linking many nutrients, foods, and dietary patterns to the risk of stroke.


Hospital admissions for respiratory system diseases in adults with intellectual disabilities in Southeast London: a register-based cohort study.
Intellectual disability (ID) carries a high impact on need for care, health status and premature mortality. Respiratory system diseases contribute a major part of mortality among people with ID, but remain underinvestigated as consequent morbidities. Anonymised electronic mental health records from the South London and Maudsley Trust (SLaM) were linked to national acute medical care data. Using retrospective cohort and matched case-control study designs, adults with ID receiving SLaM care between 1 January 2008 and 31 March 2013 were identified and compared with local catchment residents for respiratory system disease admissions. Standardised admission ratios (SARs) were first calculated, followed by a comparison of duration of hospitalisation with respiratory system disease between people with ID and age-matched and gender-matched random counterparts modelled using linear regression. Finally, the risk of readmission for respiratory system disease was analysed using the Cox models. For the 3138 adults with ID identified in SLaM, the SAR for respiratory system disease admissions was 4.02 (95% CI 3.79 to 4.26). Compared with adults without ID, duration of hospitalisation was significantly longer by 2.34 days (95% CI 0.03 to 4.64) and respiratory system disease readmission was significantly elevated (HR=1.35; 95% CI 1.17 to 1.56) after confounding adjustment. Respiratory system disease admissions in adults with ID are more frequent, of longer duration and have a higher likelihood of recurring. Development and evaluation of potential interventions to the preventable causes of respiratory diseases should be prioritised.


GFP-aided confocal laser scanning microscopy can monitor Agrobacterium tumefaciens cell morphology and gene expression associated with infection.
We tagged Agrobacterium tumefaciens cells with a mini-Tn5 transposon containing a promoterless gene encoding a green fluorescent protein (GFP). Some of the GFP-tagged individual bacterial cells exhibited strong green fluorescence, which reflected the expression levels of the GFP-tagged genes. Those cells could be readily detected with a confocal laser scanning microscope (CLSM). We observed that the fluorescence and morphology of A. tumefaciens cells grown in plant tissues resembled those grown in a minimal medium of low pH, which is required for expression of the virulence genes responsible for tumorigenesis. This suggests that GFP-aided CLSM can be used to determine which growth medium is more representative of the nutritional conditions that a pathogen encounters in plant tissues. We also observed that the fluorescence and morphology of A. tumefaciens cells changed dramatically during the course of infection. Our data suggested that A. tumefaciens cells were probably better fed upon successful colonization. We believe that GFP-aided CLSM can help study the fate of A. tumefaciens cells inside plant tissues by monitoring cell morphology and gene expression associated with the infection process in situ.


Health care and community-based interventions for war-traumatized people in Croatia: community-based study of service use and mental health.
To explore the use of health care and community-based services in war-affected regions of Croatia and its relation to mental health. A sample of 719 adults exposed to at least one war-related traumatic event were selected by random-walk technique from three Croatian counties and interviewed for socio-demographic data, mental health status (Mini International Neuropsychiatric Interview), and service use (Matrix for the Assessment of Community and Healthcare Services) in the period from 1991 to 2006. Descriptive analysis of service use was performed. Relations between service use, current mental health, and recovery from posttraumatic stress disorder (PTSD) were analyzed using logistic regression models. The traumatized population used a wide range of health care and community-based services. Health care was the most frequently used service category, especially primary health care (92.5%), followed by accommodation support (57.9%), financial support (57.7%), and employment support (32.5%). Compared with participants without mental disorders, participants with current PTSD were more likely to use only legal support (odds ratio [OR], 2.15; 95% confidence interval [CI], 1.15-3.99), while participants with other mental disorders were more likely to use social support and contacts (OR, 1.72; 95% CI, 1.08-2.75). Receiving accommodation support (OR, 2.05; 95% CI, 1.03-4.06) was the only significant predictor of recovery from PTSD, while seeking legal support (OR, 0.28; 95% CI, 0.08-0.92) was related to slower recovery. Although a wide range of services were organized to help the traumatized population in Croatia, only the solution of housing issue significantly predicted recovery. The organization of help services should take into consideration the existing infrastructure and local specificities, and respect the needs of people in war-affected areas.


3D Numerical Modeling of Laser Assisted Tape Winding Process of Composite Pressure Vessels and Pipes-Effect of Winding Angle, Mandrel Curvature and Tape Width.
Advanced thermoplastic composites manufacturing using laser assisted tape placement or winding (LATP/LATW) is a challenging task as monitoring and predicting nip point (bonding) temperature are difficult especially on curved surfaces. A comprehensive numerical analysis of the heat flux and temperature distribution near the nip point is carried out in this paper for helical winding of fiber reinforced thermoplastic tapes on a cylindrically shaped mandrel. An optical ray-tracing technique is coupled with a numerical heat transfer model in the process simulation tool. The developed optical-thermal model predictions were compared with experimental data available in literature to validate its effectiveness. The influences of winding/placement angle, mandrel curvature and tape width on the incident angles, the laser absorbed intensity, and the process temperature distribution are studied extensively using the validated model. Winding/placement angle has a considerable effect on the temperature distribution. Increase in winding angle results in a higher temperature for tape due to more reflections coming from the substrate. On the other hand, substrate temperature decreases as the winding angle increases due to a decrease in the laser incident angles based on the local surface curvature. An increase in mandrel curvature results in higher nip point temperatures for substrate and lower one for tape. Different mandrel sizes for 90 ∘ placement path do not have a strong effect on the substrate process temperature as for other winding angles because of less curvature change of the corresponding irradiated area. Tape width causes local temperature variations at the edges of the tape/substrate. In order to obtain the desired process temeprature during LATW or LATP processes, the laser intensity distribution on the tape and substrate surfaces should be regulated.


Identification of a low molecular weight inhibitor of osteoblast mitogenesis in uremic plasma.
A low molecular weight inhibitor of cartilage sulfation has been detected in the plasma of dialysis patients. Preliminary studies of this inhibitor have suggested that it may have a role in decreasing bone mass, possibly by suppressing bone cell proliferation. Since the in vitro bioassay of crude sulfation inhibitor preparations is relatively nonspecific, we investigated whether there might also be an inhibitor of osteoblast mitogenesis in uremic plasma. We fractionated plasma and plasma ultrafiltrates from dialysis patients by gel filtration chromatography and looked for inhibition of mitogenesis in cultured osteoblasts. Material from fractions with a molecular weight range of 750 to 900 inhibited osteoblast mitogenesis. The inhibitory effect, however, could be overcome with serum or insulin-like growth factor-I, suggesting that the mechanism of inhibition was not growth factor dependent. Further characterization of the inhibitor revealed that it was not a peptide or a polar lipid. We conclude that uremic plasma contains a bone cell mitogenic inhibitor which may have a role in regulating bone remodeling in adults and bone growth in children.


Water and elevation are more important than burn severity in predicting bat activity at multiple scales in a post-wildfire landscape.
Bats are among the most widespread mammals on Earth, and are subject to habitat change, loss, and other disturbances such as fire. Wildfire causes rapid changes in vegetation that affect habitat use. However, the spatial scale at which these changes affect bats depends on their use of habitat elements. Three years post wildfire, we assessed how burn severity, water, landform type, elevation, vegetation type, and roads affected use by bats of a forest landscape at multiple spatial scales. We deployed acoustic detectors at randomly selected locations within a 217,712 ha wildfire boundary in Arizona. We classified echolocation calls to species or group and calculated an activity index by adjusting the calls per hour. We conducted a multi-scale analysis of landscape structure and composition around each location from a 90 to 5760 m radius. No scale was selected preferentially by any species or group. Stream density and elevation range were more important predictors for species groups than burn severity. When burn severity was a predictor, agile species had higher activity in areas that were unburned or had low severity burn. A heterogeneous landscape composed of high, medium, and low burn severity patches within a forest altered by large wildfires provided habitat for different bat species, but water density and range in elevation were more important for predicting bat habitat use than fire severity in this arid landscape. More than one spatial scale, representing local to landscape levels, should be considered in managing habitat for bats. In arid areas, such as the western United States, maintaining reliable water sources is important for bats. Managing these factors at multiple spatial scales will benefit bat species with different wing morphologies, echolocation call types, and habitat selections.


Insect odour perception: recognition of odour components by flower foraging moths.
Odours emitted by flowers are complex blends of volatile compounds. These odours are learnt by flower-visiting insect species, improving their recognition of rewarding flowers and thus foraging efficiency. We investigated the flexibility of floral odour learning by testing whether adult moths recognize single compounds common to flowers on which they forage. Dual choice preference tests on Helicoverpa armigera moths allowed free flying moths to forage on one of three flower species; Argyranthemum frutescens (federation daisy), Cajanus cajan (pigeonpea) or Nicotiana tabacum (tobacco). Results showed that, (i) a benzenoid (phenylacetaldehyde) and a monoterpene (linalool) were subsequently recognized after visits to flowers that emitted these volatile constituents, (ii) in a preference test, other monoterpenes in the flowers' odour did not affect the moths' ability to recognize the monoterpene linalool and (iii) relative preferences for two volatiles changed after foraging experience on a single flower species that emitted both volatiles. The importance of using free flying insects and real flowers to understand the mechanisms involved in floral odour learning in nature are discussed in the context of our findings.


Social network analysis of physician interactions: the effect of institutional boundaries on breast cancer care.
Although comparative effectiveness trials and nationally recognized clinical guidelines offer substantial guidance about ideal patient treatment, we remain largely uninformed about the patterns of care seen in everyday clinical practice. To address this gap in knowledge, we looked at registry-based data on breast cancer care at two neighboring healthcare institutions with a specific focus on whether organizational boundaries determine the physicians that a patient will see. From an initial patient-oriented data set, we developed a social network of physicians, modeling their interactions over the course of the provided treatments. Applying a mixture of visual and quantitative analyses to this network, we found evidence for strong intra-institutional ties and poignantly weak connections to physicians operating out of other healthcare centers.


Developing a Conversation Aid to Support Shared Decision Making: Reflections on Designing Anticoagulation Choice.
Patient-centered care requires that treatments respond to the problematic situation of each patient in a manner that makes intellectual, emotional, and practical sense, an achievement that requires shared decision making (SDM). To implement SDM in practice, tools-sometimes called conversation aids or decision aids-are prepared by collating, curating, and presenting high-quality, comprehensive, and up-to-date evidence. Yet, the literature offers limited guidance for how to make evidence support SDM. Herein, we describe our approach and the challenges encountered during the development of Anticoagulation Choice, a conversation aid to help patients with atrial fibrillation and their clinicians jointly consider the risk of thromboembolic stroke and decide whether and how to respond to this risk with anticoagulation.


Sensory cues for perceived exertion: a review.
A variety of physiological responses, classified as local or central factors, have been suggested as providing the primary input for perception of effort during exercise. The potency of these responses as perceptual cues, however, has seldom been addressed in terms of relevant modifying variables such as exercise intensity, exercise duration, exercise modality, and steady-state vs progressive exercise. This paper provides a critical review of the most relevant perceived exertion literature in terms of these considerations. It has been proposed that the impact of physiological responses as perceptual cues is dependent on their availability to conscious monitoring during exercise. The viability of this suggestion as a criterion for determining the importance of local and central cues for perceived exertion is also examined.


Ruminant-produced trans-fatty acids raise plasma HDL particle concentrations in intact and ovariectomized female Hartley guinea pigs.
Cardiovascular disease (CVD) is the leading cause of death among women worldwide, and risk for developing CVD increases postmenopause. Consumption of trans-fatty acids (tFA) has been positively associated with CVD incidence and mortality. The current study was designed to assess the effects of diets high in industrially produced (IP)-tFA, from partially hydrogenated vegetable oils (PHVO), and ruminant-produced (RP)-tFA, from butter oil (BO), on risk factors for CVD. Thirty-two female Hartley guinea pigs, one-half of which were ovariectomized (OVX) to mimic the postmenopausal condition, were fed hypercholesterolemic diets containing 9% by weight PHVO or BO (n = 8/diet and ovariectomy) for 8 wk. The plasma and hepatic lipids did not differ between IP- and RP-tFA groups or between intact and OVX guinea pigs. The BO diet resulted in higher concentrations of plasma total and small HDL particle subclass concentrations than the PHVO diet regardless of ovariectomy status. The intact BO group had higher concentrations of large HDL particles than the intact PHVO group. HDL mean particle size tended to be larger (P = 0.07) in the PHVO groups compared with the BO groups regardless of ovariectomy status. There was a trend toward an interaction between diet and ovariectomy status for LDL mean particle size, which tended to be larger in OVX guinea pigs fed PHVO (P = 0.07). In summary, consumption of IP- and RP-tFA resulted in differential effects on HDL particle subclass profiles in female guinea pigs. The effect of tFA consumption and hormonal status on HDL particle subclass metabolism and the subsequent impact on CVD in females warrants further investigation.


Testing the distinctness of shoot ionomes of angiosperm families using the Rothamsted Park Grass Continuous Hay Experiment.
• The ionome is the elemental composition of a tissue or organism. Phylogenetic variation in the ionomes of plant shoots has been widely reported based on controlled experiments, vegetation surveys and literature meta-analyses. However, environmental effects on phylogenetic variation in shoot ionomes have not been quantified. This study tests the hypothesis that phylogenetic variation in shoot ionomes is robust to environmental perturbation and that plant families can be distinguished by their shoot ionomes. • Herbage was sampled from six subplots of the Rothamsted Park Grass Experiment. Subplots had received contrasting fertilizer treatments since 1856. Herbage was separated into its constituent species (n = 21) and concentrations of eleven mineral elements were determined in dried shoot material. • Shoot concentrations of calcium (Ca), zinc (Zn), manganese (Mn), magnesium (Mg) and sodium (Na) showed significant variation associated with plant species, and responded similarly to fertilizer treatments in diverse plant species. Species × treatment interactions were indicated for phosphorus (P), potassium (K), nickel (Ni), copper (Cu) and iron (Fe). Plant families could be distinguished by their shoot ionomes. The most informative elements for discriminant analysis were Ca > Mg > Ni > S > Na > Zn > K > Cu > Fe > Mn > P. • Whilst shoot ionomes were sensitive to fertilizer treatment, phylogenetic variation in a subset of the shoot ionome (Ca, Zn, Mn, Mg) was robust to this environmental perturbation.


Devil's Claw to suppress appetite--ghrelin receptor modulation potential of a Harpagophytum procumbens root extract.
Ghrelin is a stomach-derived peptide that has been identified as the only circulating hunger hormone that exerts a potent orexigenic effect via activation of its receptor, the growth hormone secretagogue receptor (GHS-R1a). Hence, the ghrelinergic system represents a promising target to treat obesity and obesity-related diseases. In this study we analysed the GHS-R1a receptor activating potential of Harpagophytum procumbens, popularly known as Devil's Claw, and its effect on food intake in vivo. H. procumbens is an important traditional medicinal plant from Southern Africa with potent anti-inflammatory and analgesic effects. This plant has been also used as an appetite modulator but most evidences are anecdotal and to our knowledge, no clear scientific studies relating to appetite modulation have been done to this date. The ghrelin receptor activation potential of an extract derived from the dried tuberous roots of H. procumbens was analysed by calcium mobilization and receptor internalization assays in human embryonic kidney cells (Hek) stably expressing the GHS-R1a receptor. Food intake was investigated in male C57BL/6 mice following intraperitoneal administration of H. procumbens root extract in ad libitum and food restricted conditions. Exposure to H. procumbens extract demonstrated a significant increased cellular calcium influx but did not induce subsequent GHS-R1a receptor internalization, which is a characteristic for full receptor activation. A significant anorexigenic effect was observed in male C57BL/6 mice following peripheral administration of H. procumbens extract. We conclude that H. procumbens root extract is a potential novel source for potent anti-obesity bioactives. These results reinforce the promising potential of natural bioactives to be developed into functional foods with weight-loss and weight maintenance benefits.


Vitamin B2 intake and the risk of colorectal cancer: a meta-analysis of observational studies.
A systematic review and meta-analysis of observational studies evaluated the association of intake of vitamin B2 with the incidence of colorectal cancer. Relevant studies were identified in MEDLINE via PubMed (published up to April 2014). We extracted data from articles on vitamin B2 and used multivariable-adjusted odds ratio (OR) and a random-effects model for analysis. We found 8 articles meeting the inclusion criteria (4 of cohort studies and 4 of case-control studies) and a total of 7,750 colorectal cancer cases were included in this meta-analysis. The multivariable-adjusted OR for pooled studies for the association of the highest versus lowest vitamin B2 intake and the risk of colorectal cancer was 0.83 (95% confidence interval [95%CI]:0.75,0.91). We performed a sensitivity analysis for vitamin B2. If we omitted the study by Vecchia et al., the pooled OR was 0.86 (95%CI, 0.77,0.96). This is the first meta-analysis to study links between vitamin B2 and colorectal cancer. We found vitamin B2 intake was inversely associated with risk of colorectal cancer. However, further research and large sample studies need to be conducted to better validate the result.


Histological and histochemical study of the vesicoureteric junction in infancy and childhood.
The morphology of the vesicoureteric junction was compared using 19 post mortem specimens obtained from male and female Afro-American and Caucasian children with an age range of 1 to 72 months (mean 4). All specimens were serially sectioned and the tissues processed using standard histological and histochemical techniques (acetylcholinesterase ¿AChE¿ and pseudocholinesterase ¿PChE¿). The results failed to reveal any differences in the structure of the vesicoureteric junction with respect to age, sex and ethnic origins. The vesicoureteric junction comprised 3 histologically and histochemically distinct smooth muscle components. Ureteric muscle formed a complete inner layer rich in PChE which continued beyond the ureteric orifices to merge distally with the superficial trigone. An intermediate layer of muscle was also demonstrated whose constituent muscle cells possessed specific histological features and which was rich in both AChE and PChE, which is distinct from that derived from the ureter and detrusor. The presence of detrusor muscle on the outer aspect of the juxtavesical segment of ureter rich in AChE was also confirmed. While this study, using histochemical studies in infants and children, did not reveal any differences in the structure of the vesicoureteric junction with respect to age, sex or ethnic origin, an intermediate layer of muscle was identified with histochemical characteristics more like that of the male genital tract than that derived from the ureter or detrusor muscle.


[Effect of insulin and hydrocortisone on the isoenzyme composition of pyruvate kinase in the rabbit kidney].
Alloxan diabetes, starvation of rabbits within one day and administration of hydrocortisone during 5 days did not cause distinct alterations in the total activity of pyruvate kinase in kidney cortex. At the same time pronounced alterations in the isozyme spectra were observed: the L-type activity of pyruvate kinase was decreased and the M2-type of activity was increased. The total activity and the isozyme content of pyruvate kinase were only slightly altered in starvation of rabbits during 3 or 10-16 days and after administration of protamine-Zn-insulin during 3 days. Hydrocortisone caused variable effects on the pyruvate kinase isozymes from kidney cortex, depending on periods of administration.


Synthesis of N-Doped Magnetic WO3-x@Mesoporous Carbon Using a Diatom Template and Plasma Modification: Visible-Light-Driven Photocatalytic Activities.
Synthesis of three-dimensional photocatalysts offers great potential for chemical conversion and hydrogen generation as appropriate solutions for environmental protection and energy shortage challenges. In this study, the magnetic WO3-x@mesoporous carbon (M-WO3-x@MC) was synthesized through the evaporation-induced self-assembly method applying diatom frustules as a natural template. Then, plasma modification was used to prepare the N-doped M-WO3-x@MC (NM-WO3-x@MC) with enhanced photocatalytic activity and durable performance. The WO3-x was embedded in the conductive MC, which was also partially reduced by the carbon precursor within the heat-treatment procedure. The obtained M-WO3-x@MC was treated by the plasma under an N2 atmosphere for the production of the final photocatalyst containing both the N-doped WO3-x and MC. As a result, the NM-WO3-x@MC had larger surface area (208.4 m2 g-1), narrower band gap (2.3 eV), more visible light harvesting, and confined electron-hole pairs recombination. The H2 generation rates of net WO3 nanorods and NM-WO3-x@MC nanocomposite were estimated as 532 and 2765 μmol g-1 h-1, respectively. Additionally, more than 90% of antibiotics (cephalexin, cefazolin and cephradine) degradation and 76% of total organic carbon elimination were obtained after 120 and 240 min of photocatalytic process under visible light irradiation. Eventually, more than eight intermediates were detected for each antibiotic degradation using the gas chromatography-mass spectrometer method, and based on the obtained results, the possible degradation pathways were suggested.


Balloon-tipped catheter arterial occlusion for prevention of hemorrhage caused by guttural pouch mycosis: 13 cases (1982-1985).
Epistaxis, caused by guttural pouch mycosis, was treated by balloon-tipped catheter and ligature occlusion of the involved arteries in 13 horses. In 7 horses, more than one artery was catheterized. Serious postoperative hemorrhage was prevented in all horses. Long-term endoscopic follow-up examination was possible in 8 horses, and all had complete regression of the fungal lesion as early as 5 weeks after surgery. Six of the 8 horses did not receive medical treatment. Problems related to the use of the balloon-tipped catheter technique were rare. There were incisional infections in 3 horses, catheter breakage during removal occurred twice, and in one horse, a catheter was inappropriately placed.


Energy drinks: an emerging public health hazard for youth.
Energy drinks are emerging as a public health threat and are increasingly consumed by youth internationally. Energy drinks contain high levels of caffeine, sugar, and novel ingredients, and are often marketed through youth-oriented media and venues. We review these practices and the current inconsistent state of labeling. We also examine international support for regulation of these products, including a survey showing that 85 per cent of United States parents agreed that regulations requiring caffeine content disclosure and warning labels on energy drinks are warranted. We then examine the regulatory structure for energy drinks in the United States, analyzing legal and self-regulatory strategies to protect consumers, especially youth, from these potentially dangerous products. Recommended government interventions include revised labeling requirements, addressing problematic ingredients, and enacting retail restrictions. We conclude by identifying areas for future research.


JSEM: a framework for identifying and evaluating indicators.
There are two issues in indicator development that have not been adequately addressed: (1) how to select an optimal combination of potentially redundant indicators that together best represent an endpoint, given cost constraints; (2) how to identify and evaluate indicators when the endpoint is unmeasured. This paper presents an approach to identifying and evaluating combinations of indicators when the mathematical relationships between the indicators and an endpoint may not be quantified, a limitation common to many ecological assessments. The approach uses the framework of Structural Equation Modeling (SEM), which combines path analysis with measurement models, to formalize available information about potential indicators and to evaluate their potential adequacy for representing an endpoint. Unlike traditional applications of SEM which require data on all variables, our approach---judgement-based SEM (JSEM)--can utilize expert judgement regarding the strengths and shapes of indicator-endpoint relationships. JSEM is applied in two stages. First, a conceptual model that relates variables in a network of direct and indirect linkages is developed, and is used to identify indicators relevant to an endpoint. Second, an index of indicator strength--i.e., the strength of the relationship between the endpoint and a set of indicators--is calculated from estimates of correlation between the modeled variables, and is used to compare alternative sets of indicators. The second stage is most appropriate for large, long-term assessments. Although JSEM is not a statistical technique, basing JSEM on SEM provides a structure for validating the conceptual model and for relining the index of indicator strength as data become available. Our main objective is to contribute to a rigorous and consistent selection of indicators even when knowledge about the ability of indicators to represent an endpoint is limited to expert judgement.


Sexual Dysfunction and Gun Ownership in America: When Hard Data Meet a Limp Theory.
Although there has been no direct empirical evidence linking sexual dysfunction (SD) with gun ownership, speculation has been widespread and persistent for decades. In this paper, we formally examine the association between SD and gun ownership. Our primary hypothesis, derived from the psychosexual theory of gun ownership, asserts that men experiencing SD are more likely to personally own guns than other men. To test this hypothesis, we used recently collected data from the 2021 Crime, Health, and Politics Survey (CHAPS), a national probability sample of 780 men, and binary logistic regression to model gun ownership as a function of SD. Our key finding is that men experiencing SD are no more likely to own guns than men without SD. This interpretation was supported across several indicators of SD (performance anxiety, erection trouble, and ED medication) and gun ownership (personal gun ownership, purchasing a gun during the pandemic, and keeping a gun in one's bedroom). To our knowledge, we are the first to have directly tested the association between SD and gun ownership in America. Our findings are important because they contribute to our understanding of factors associated with gun ownership by challenging the belief that phallic symbolism and masculinity somehow drive men with SD to purchase guns. Our results also remind us of the perils of gun culture rhetoric, which, in this case, function to discredit gun owners and to further stigmatize men with ED. We conclude by calling for more evidence-based discussions of SD and guns in society.


Total cortisol output during an acute stressor predicts youths' internalizing symptoms during the COVID-19 pandemic.
Individual differences in cortisol output may influence adolescents' adjustment to the COVID-19 pandemic; however, boys and girls may differ in terms of associations between cortisol output and internalizing symptoms in the context of COVID-19-related stress. We examined whether pre-pandemic cortisol output during an acute stressor, assessed approximately three years prior to the pandemic, predicted change in adolescents' internalizing symptoms early during the COVID-19 pandemic. Consistent with previous work on other life stressors, girls' cortisol output was positively associated with anxious and somatic symptoms early in the pandemic. Conversely, cortisol output and depressive symptoms were negatively associated for boys; boys with higher cortisol had depressive symptoms which significantly decreased over time. Findings suggest that hypothalamic-pituitary-adrenal axis stress functioning plays a role in shaping differences between adolescent boys' and girls' adjustment during the experience of a ubiquitous chronic stressor.


[One-stage formation of urinary bladder, restoration of symphysis with correction of congenital defects of external and internal genital organs].
One-stage combined surgery in a female patient with total extrophy of the urinary bladder after uretherosigmostomy was performed for the first time with participation of surgeons, gynecologists and urologists. Corrective surgeries in these patients must be performed for the purpose of creation of isolated urinary reservoir for prophylaxis of pyelonephritis and chronic renal insufficiency. Correction of symphysis defect was carried out with polypropylene endograft. This surgery was performed to provide quality of life and reproductive function of the patient.


Observations on the physiology of adult anophelines in Asia.
Some recent physiological observations made in Afghanistan, Ceylon and Burma on adults of Anopheles culicifacies and A. aconitus are described. The specimens from Ceylon and Burma, in particular, showed a peculiar physiological process in which, at the beginning of the dry season, the coagulated, dark red, and almost desiccated stomach contents indicated a period of partial quiescence. As there were in most cases no signs of apparent growth of the fat-body, it appeared that a hitherto undescribed form of physiological behaviour occurred under certain climatic and seasonal conditions.


Ursolic acid inhibits early lesions of diabetic nephropathy.
The present study sought to investigate the effects of ursolic acid (UA) on the development of glomerular hypertrophy and type IV collagen accumulation, two early lesions associated with diabetic nephropathy (DN). By treating streptozotocin (STZ)-induced diabetic mice with low-dose UA (0.01% in food) for three months, the diabetes-induced glomerular hypertrophy and type IV collagen accumulation in the kidneys were found to be markedly ameliorated. Further studies identified that UA treatment suppressed diabetes-induced activations of STAT-3, ERK1/2 and JNK pathways, but not the diabetes-induced activation of the p38 pathway. Furthermore, diabetes-induced overexpression of iNOS in the renal cortex was also significantly suppressed by the treatment. UA may thus be considered as a potential therapeutic agent in treating DN.


COVID-RO study: the radiation oncology practice at times of COVID-19 outbreak - international survey.
Radiation therapy (RT), an essential treatment of cancer, involves multiple hospital visits. We hypothesized that radiation departments would adjust their work patterns and RT protocols in response to the SARS-CoV-2 pandemic. An electronic survey was sent during April 2020 to an international sample of radiation oncologists. The survey explored various aspects of departmental preparedness, and changes to their institutional RT protocols. A total of 68 radiation oncologists from 13 countries answered the survey. Healthcare systems were at least moderately affected in 76%. Most institutes appeared well prepared for the outbreak: regarding the availability of personal protective equipment, tests, and telemedicine/videoconference facilities. Screening for SARS-CoV-2 was applied in 59% of responders. Modification of RT protocols were minor in 66%, significant in 19% and no changes made in 15%. The extent to which protocols were modified correlated with overall healthcare disruption (p = 0.028). Normal fractionation was recommended to continue in 83% and 85% of head & neck, and cervical cancers vs. 64% of lung cancers (p = 0.001).In case the pandemic worsens, there was strong agreement to prioritize RT for aggressive cancers (80%), delay RT for slow-growing tumors (78%) and change to evidance-based hypofractionations protocols (79.4%). The option of delayed/omitted adjuvant RT (not site specific) was selected in 47%. This international survey concludes that, by making significant organizational adjustments and minor protocol modifications, RT may be safely continued during this pandemic. If the crisis worsens, there was strong agreement to continue the treatment of aggressive tumors and utilize evidence-based hypofractionated protocols.


Degradation in landscape matrix has diverse impacts on diversity in protected areas.
A main goal of protected areas is to maintain species diversity and the integrity of biological assemblages. Intensifying land use in the matrix surrounding protected areas creates a challenge for biodiversity conservation. Earlier studies have mainly focused on taxonomic diversity within protected areas. However, functional and especially phylogenetic diversities are less studied phenomena, especially with respect to the impacts of the matrix that surrounds protected areas. Phylogenetic diversity refers to the range of evolutionary lineages, the maintenance of which ensures that future evolutionary potential is safeguarded. Functional diversity refers to the range of ecological roles that members of a community perform. For ecosystem functioning and long-term resilience, they are at least as important as taxonomic diversity. We studied how the characteristics of protected areas and land use intensity in the surrounding matrix affect the diversity of bird communities in protected boreal forests. We used line-transect count and land-cover data from 91 forest reserves in Northern Finland, and land-cover data from buffer zones surrounding these reserves. We studied if habitat diversity and productivity inside protected areas, and intensity of forest management in the matrix have consistent effects on taxonomic, functional and phylogenetic diversities, and community specialization. We found that habitat diversity and productivity inside protected areas have strong effects on all diversity metrics, but matrix effects were inconsistent. The proportion of old forest in the matrix, reflecting low intensity forest management, had positive effects on community specialization. Interestingly, functional diversity increased with increasing logging intensity in the matrix. Our results indicate that boreal forest reserves are not able to maintain their species composition and abundances if embedded in a severely degraded matrix. Our study also highlights the importance of focusing on different aspects of biodiversity.


Heparan sulfate proteoglycan and laminin mediate two different types of neurite outgrowth.
Spinal cord neurons cultured in vitro have been shown to respond to changes in their environment by means of 2 different types of neurite outgrowth: (1) neurite elongation and (2) emergence and branching of newly formed neurites. Culture of spinal cord neurons with heparan sulfate proteoglycan (HSPG) medium resulted in a 3-fold increase in neurite elongation compared to the control. Extensive branching was seen when neurons were cultured in laminin-supplemented culture medium. HSPG-induced elongation and laminin-induced branching of neurites were blocked by specific anti-HSPG and antilaminin sera, respectively. Furthermore, laminin antibodies did not inhibit neurite elongation and HSPG antibodies did not block neurite branching. Conditioned medium from primary embryonic rat muscle cultures (MCM) mimicked the effects of both HSPG and laminin on neurite outgrowth. Immunoprecipitation with anti-HSPG and antilaminin antibodies demonstrated that MCM contains these 2 basal lamina components. Our observations suggest that HSPG and laminin might be highly effective molecules for promoting neurite outgrowth of rat spinal cord neurons in vitro.


Revisiting the protein-coding gene catalog of Drosophila melanogaster using 12 fly genomes.
The availability of sequenced genomes from 12 Drosophila species has enabled the use of comparative genomics for the systematic discovery of functional elements conserved within this genus. We have developed quantitative metrics for the evolutionary signatures specific to protein-coding regions and applied them genome-wide, resulting in 1193 candidate new protein-coding exons in the D. melanogaster genome. We have reviewed these predictions by manual curation and validated a subset by directed cDNA screening and sequencing, revealing both new genes and new alternative splice forms of known genes. We also used these evolutionary signatures to evaluate existing gene annotations, resulting in the validation of 87% of genes lacking descriptive names and identifying 414 poorly conserved genes that are likely to be spurious predictions, noncoding, or species-specific genes. Furthermore, our methods suggest a variety of refinements to hundreds of existing gene models, such as modifications to translation start codons and exon splice boundaries. Finally, we performed directed genome-wide searches for unusual protein-coding structures, discovering 149 possible examples of stop codon readthrough, 125 new candidate ORFs of polycistronic mRNAs, and several candidate translational frameshifts. These results affect >10% of annotated fly genes and demonstrate the power of comparative genomics to enhance our understanding of genome organization, even in a model organism as intensively studied as Drosophila melanogaster.


Multi-target-directed phenol-triazole ligands as therapeutic agents for Alzheimer's disease.
Alzheimer's disease (AD) is a multifactorial disease that is characterized by the formation of intracellular neurofibrillary tangles and extracellular amyloid-β (Aβ) plaque deposits. Increased oxidative stress, metal ion dysregulation, and the formation of toxic Aβ peptide oligomers are all considered to contribute to the etiology of AD. In this work we have developed a series of ligands that are multi-target-directed in order to address several disease properties. 2-(1-(3-Hydroxypropyl)-1H-1,2,3-triazol-4-yl)phenol (POH), 2-(1-(2-morpholinoethyl)-1H-1,2,3-triazol-4-yl)phenol (PMorph), and 2-(1-(2-thiomorpholinoethyl)-1H-1,2,3-triazol-4-yl)phenol (PTMorph) have been synthesized and screened for their antioxidant capacity, Cu-binding affinity, interaction with the Aβ peptide and modulation of Aβ peptide aggregation, and the ability to limit Aβ1-42-induced neurotoxicity in human neuronal culture. The synthetic protocol and structural variance incorporated via click chemistry, highlights the influence of R-group modification on ligand-Aβ interactions and neuroprotective effects. Overall, this study demonstrates that the phenol-triazole ligand scaffold can target multiple factors associated with AD, thus warranting further therapeutic development.


Is the Preoperative Use of Antidepressants and Benzodiazepines Associated with Opioid and Other Analgesic Use After Hip and Knee Arthroplasty?
Mental health disorders can occur in patients with pain conditions, and there have been reports of an increased risk of persistent pain after THA and TKA among patients who have psychological distress. Persistent pain may result in the prolonged consumption of opioids and other analgesics, which may expose patients to adverse drug events and narcotic habituation or addiction. However, the degree to which preoperative use of antidepressants or benzodiazepines is associated with prolonged analgesic use after surgery is not well quantified. (1) Is the preoperative use of antidepressants or benzodiazepine medications associated with a greater postoperative use of opioids, NSAIDs, or acetaminophen? (2) Is the proportion of patients still using opioid analgesics 1 year after arthroplasty higher among patients who were taking antidepressants or benzodiazepine medications before surgery, after controlling for relevant confounding variables? (3) Does analgesic drug use decrease after surgery in patients with a history of antidepressant or benzodiazepine use? (4) Does the proportion of patients using antidepressants or benzodiazepines change after joint arthroplasty compared with before? Of the 10,138 patients who underwent hip arthroplasty and the 9930 patients who underwent knee arthroplasty at Coxa Hospital for Joint Replacement, Tampere, Finland, between 2002 and 2013, those who had primary joint arthroplasty for primary osteoarthritis (64% [6502 of 10,138] of patients with hip surgery and 82% [8099 of 9930] who had knee surgery) were considered potentially eligible. After exclusion of another 8% (845 of 10,138) and 13% (1308 of 9930) of patients because they had revision or another joint arthroplasty within 2 years of the index surgery, 56% (5657 of 10,138) of patients with hip arthroplasty and 68% (6791 of 9930) of patients with knee arthroplasty were included in this retrospective registry study. Patients who filled prescriptions for antidepressants or benzodiazepines were identified from a nationwide drug prescription register, and information on the filled prescriptions for opioids (mild and strong), NSAIDs, and acetaminophen were extracted from the same database. For the analyses, subgroups were created according to the status of benzodiazepine and antidepressant use during the 6 months before surgery. First, the proportions of patients who used opioids and any analgesics (that is, opioids, NSAIDs, or acetaminophen) were calculated. Then, multivariable logistic regression adjusted with age, gender, joint, Charlson Comorbidity Index, BMI, laterality (unilateral/same-day bilateral), and preoperative analgesic use was performed to calculate odds ratios for any analgesic use and opioid use 1 year postoperatively. Additionally, the proportion of patients who used antidepressants and benzodiazepines was calculated for 2 years before and 2 years after surgery. At 1 year postoperatively, patients with a history of antidepressant or benzodiazepine use were more likely to fill prescriptions for any analgesics than were patients without a history of antidepressant or benzodiazepine use (adjusted odds ratios 1.9 [95% confidence interval 1.6 to 2.2]; p < 0.001 and 1.8 [95% CI 1.6 to 2.0]; p < 0.001, respectively). Similarly, patients with a history of antidepressant or benzodiazepine use were more likely to fill prescriptions for opioids than patients without a history of antidepressant or benzodiazepine use (adjusted ORs 2.1 [95% CI 1.7 to 2.7]; p < 0.001 and 2.0 [95% CI 1.6 to 2.4]; p < 0.001, respectively). Nevertheless, the proportion of patients who filled any analgesic prescription was smaller 1 year after surgery than preoperatively in patients with a history of antidepressant (42% [439 of 1038] versus 55% [568 of 1038]; p < 0.001) and/or benzodiazepine use (40% [801 of 2008] versus 55% [1098 of 2008]; p < 0.001). The proportion of patients who used antidepressants and/or benzodiazepines was essentially stable during the observation period. Surgeons should be aware of the increased risk of prolonged opioid and other analgesic use after surgery among patients who were on preoperative antidepressant and/or benzodiazepine therapy, and they should have candid discussions with patients referred for elective joint arthroplasty about this possibility. Further studies are needed to identify the most effective methods to reduce prolonged postoperative opioid use among these patients. Level III, therapeutic study.


Beyond Negative Affectivity: A Hierarchical Model of Global and Transdiagnostic Vulnerabilities for Emotional Disorders.
Negative affectivity (NA) has been linked to anxiety and depression (DEP). Identifying the common factors between anxiety and DEP is important when explaining their overlap and comorbidity. However, general factors such as NA tend to have differential relationships with different disorders, suggesting the need to identify mediators in order to explicate these relationships. The current study tests a theoretically and empirically derived hierarchical model of emotional disorders including both a general factor (NA) and transdiagnostic risk factors [anxiety sensitivity (AS) and intolerance of uncertainty (IoU)] using structural equation modeling. AS was tested as a mid-level factor between NA and panic disorder/agoraphobia, while IoU was tested as a mid-level factor between NA and social phobia, generalized anxiety disorder, obsessive-compulsive disorder, and DEP. Data from 642 clinical outpatients with a heterogeneous presentation of emotional disorders were available for analysis. The hierarchical model fits the data adequately. Moreover, while a simplified model removing AS and IoU fits the data well, it resulted in a significant loss of information for all latent disorder constructs. Data were unavailable to estimate post-traumatic stress disorder or specific phobias. Future work will need to extend to other emotional disorders. This study demonstrates the importance of both general factors that link disorders together and semi-specific transdiagnostic factors partially explaining their heterogeneity. Including these mid-level factors in hierarchical models of psychopathology can help account for additional variance and help to clarify the relationship between disorder constructs and NA.


Proteins of the nuclear factor-1 family act as an activator of the late promoter in human polyomavirus BK in vitro.
The cis-acting elements for the early and late promoters, as well as the enhancer in the prototype strains of human polyomavirus BK (BKV) are located within a 500 bp intergenic region. We previously studied the specificity of protein binding in this region in vitro and showed that the interaction of proteins of the nuclear factor-1 (NF-1) family is crucial for early promoter activity. We have now extended our study to the BKV late promoter. We show that the late promoter activity in HeLa cell extracts is poor compared to the activity of the early promoter. Using a high template to protein ratio, multiple start sites were detected by primer extension analysis. DNase I protection experiments revealed the presence of three NF-1 binding sites in the late side, in addition to those identified previously in the 68 bp repeats and C element. Competition transcription assays using binding sites for NF-1, AP-1, Sp-1 and a complete 68 bp repeat indicated that only the 68 bp repeat and the NF-1 binding site competed significantly with the late promoter activity. A point mutation in the NF-1 binding site, which destroys the ability of the oligonucleotide to bind NF-1, also impaired its capacity to compete with the late promoter. The ability of NF-1 to activate both the early and late promoters suggests that the proteins of this family act as a bidirectional transcriptional activator in this virus.


Plasmablastic lymphoma of the small intestine: case report and literature review.
Plasmablastic lymphoma (PBL) is a rare aggressive B-cell lymphoproliferative disorder, which has been characterized by the World Health Organization as a new entity. Although PBL is most commonly seen in the oral cavity of human immunodeficiency virus (HIV)-positive patients, it can also be seen in extra-oral sites in immunocompromised patients who are HIV-negative. Here we present a rare case of PBL of the small intestine in a 55-year-old HIV-negative male. Histopathological examination of the excisional lesion showed a large cell lymphoma with plasmacytic differentiation diffusely infiltrating the small intestine and involving the surrounding organs. The neoplastic cells were diffusely positive for CD79a, CD138 and CD10 and partly positive for CD38 and epithelial membrane antigen. Approximately 80% of the tumor cells were positive for Ki-67. A monoclonal rearrangement of the kappa light chain gene was demonstrated. The patient died approximately 1.5 mo after diagnosis in spite of receiving two courses of the CHOP chemotherapy regimen. In a review of the literature, this is the first case report of PBL with initial presentation in the small intestine without HIV and Epstein-Barr virus infection, and a history of hepatitis B virus infection and radiotherapy probably led to the iatrogenic immunocompromised state.


[Multifactorial diagnosis and surgical treatment in inflammatory tumors of the uterine adnexa].
In 66 patients with a tubo-ovarian tumor of inflammatory genesis in consequence of hyperdiagnostics in gynaecological compartments of general practice net diagnosis of the ovary cancer was established. Application of complex diagnostic measures, consisting of computing diagnosis tables construction, use of straight prolonged endolymphatic antibacterial therapy, ultrasonic investigation and computer tomography as well allowed to specify the diagnosis and to conduct an adequate surgical treatment.


All-Cause and Cancer-Specific Death of Older Adults Following Surgery for Cancer.
Cancer care has inherent complexities in older adults, including balancing risks of cancer and noncancer death. A poor understanding of cause-specific outcomes may lead to overtreatment and undertreatment. To examine all-cause and cancer-specific death throughout 5 years for older adults after cancer resection. This population-based cohort study was conducted in Ontario, Canada, using the administrative databases stored at ICES (formerly the Institute for Clinical Evaluative Sciences). All adults 70 years or older who underwent resection for a new diagnosis of cancer between January 1, 2007, and December 31, 2017, were included. Patients were followed up until death or censored at date of last contact of December 31, 2018. Cancer resection. Using a competing risks approach, the cumulative incidence of cancer and noncancer death was estimated and stratified by important prognostic factors. Multivariable subdistribution hazard models were fit to explore prognostic factors. Of 82 037 older adults who underwent surgery (all older than 70 years; 52 119 [63.5%] female), 16 900 of 34 044 deaths (49.6%) were cancer related at a median (interquartile range) follow-up of 46 (23-80) months. At 5 years, estimated cumulative incidence of cancer death (20.7%; 95% CI, 20.4%-21.0%) exceeded noncancer death (16.5%; 95% CI, 16.2%-16.8%) among all patients. However, noncancer deaths exceeded cancer deaths starting at 3 years after surgery in breast, prostate, and melanoma skin cancers, patients older than 85 years, and those with frailty. Cancer type, advancing age, and frailty were independently associated with cause-specific death. At the population level, the relative burden of cancer deaths exceeds noncancer deaths for older adults selected for surgery. No subgroup had a higher burden of noncancer death early after surgery, even in more vulnerable patients. This cause-specific overall prognosis information should be used for patient counseling, to assess patterns of over- or undertreatment in older adults with cancer at the system level, and to guide targets for system-level improvements to refine selection criteria and perioperative care pathways for older adults with cancer.


The natural history, current status, and future trends of HIV infection.
Nearly 50 million people worldwide have been infected with human immunodeficiency virus (HIV), and an estimated 12 to 13 million children have been orphaned by the acquired immune deficiency syndrome (AIDS) epidemic. The natural history of HIV infection continues to evolve as researchers and clinicians unearth new facts about the virus and develop new treatment regimens for patients. The status of anti-HIV treatments is never static but constantly changing. Patients and providers struggle with adherence issues. Vaccine development, viewed as an essential step in controlling the epidemic, is complicated by the genetic diversity of the virus and the inability of the body to clear the virus. Tremendous strides have occurred in the HIV/AIDS arena, yet daunting challenges remain. This overview article presents the natural history, the current status, and future trends of HIV infection.


Second-hand smoke levels in Israeli bars, pubs and cafes before and after implementation of smoke-free legislation.
In 2007, Israel passed a law to extend existing restrictions on smoking in public places and to strengthen enforcement. Responsibility for ensuring smoke-free indoor public places was placed on establishment owners. Bars and pubs were included in the law for the first time. This study aimed to assess changes in air quality in popular Israeli bars, pubs and cafes after the implementation of law, and to examine changes in patron numbers, percentage of smoking patrons and venue-seating sections. Air quality was determined by measuring respirable suspended particles (PM(2.5) μg(-3)) in 33 randomly selected venues in Jerusalem and Tel Aviv, including bars, pubs and cafes, before and after law implementation. Numbers of patrons and smoking patrons were recorded. Average respirable small particles (RSP) level was 245 μg(-3) prior to implementation and 161 μg(-3) following implementation of the law, representing a decline of 34% (P = 0.0043). RSP levels decreased in bars and pubs and in cafes. Percentage of smoking patrons declined from 19% to 9% (P = 0.0036). The magnitude of the effect decreased over time (P = 0.0039). Non-smoking establishments were more common following the legislation (P = 0.0047). Indoor air pollution from second-hand smoke in Israeli bars, pubs and cafes in Jerusalem and Tel Aviv declined following the implementation of law. This demonstrates that a law to extend existing restrictions and enforcement policies may help protect workers and patrons from tobacco smoke. However, RSP levels in Israeli bars and pubs, especially in Tel Aviv, remain unacceptably high. Enforced, 100% smoke-free laws are essential for complete protection.


Chemical preparation and structural characterization of a homogeneous series of chitin/chitosan oligomers.
The preparation of a homogeneous series of chitin/chitosan oligomers (chito-oligomers) with the same distribution of degrees of polymerization (DP) ranging from 2 to 12, but with various average degrees of N-acetylation (DA) from 0 to 90% is described. This DA-series was obtained according to a two-step chemical process involving (i) the production of a well-defined mixture of glucosamine (GlcN) oligomers obtained by acid hydrolysis of a fully N-deacetylated chitosan and after selective precipitations of the hydrolysis products, and (ii) the partial N-acetylation of the GlcN units of these oligomers from a hydro-alcoholic solution of acetic anhydride in a controlled manner. The characterization of this series of samples with different DAs by proton nuclear magnetic resonance (1H NMR) spectroscopy and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) allowed us to determine their average DA and identify the main oligomer structures constituting each mixture. Furthermore, MALDI-TOF MS was particularly helpful to study the distribution evolution of the diverse oligomers as a function of DA for the main DPs from 3 to 7. The modeling of these distributions by means of a binomial law displayed that the chemical N-acetylation of low DP GlcN oligomers, produced in a homogeneous medium, occurs randomly along the oligosaccharide chains in accordance with a statistical (Bernoullian) arrangement. In this case, the relative proportion of each chito-oligomer present in the mixture can be estimated precisely as a function of DA considering oligomers of same DP.


Modeling the impact of universal tuberculosis molecular testing and timing of tuberculosis preventive treatment during antiretroviral therapy initiation in South Africa.
Targeted universal tuberculosis (TB) testing can improve TB detection among people with HIV. This approach is being scaled up in South Africa through Xpert MTB/RIF Ultra testing for individuals starting antiretroviral therapy and annually thereafter. Clarity is needed on how Universal Xpert testing may affect TB preventive treatment (TPT) provision, and on whether TPT should be delayed until TB is ruled out. State-transition microsimulation. We simulated a cohort of South African patients being screened for TB while entering HIV care. We compared clinical and cost outcomes between four TB screening algorithms: symptom-based, C-reactive protein-based, and Universal Xpert testing with either simultaneous or delayed TPT initiation. Prompt TB treatment initiation among simulated patients with TB increased from 26% (24-28%) under symptom screening to 53% (50-56%) with Universal Xpert testing. Universal Xpert testing led to increased TPT uptake when TPT initiation was simultaneous, but to approximately 50% lower TPT uptake if TPT was delayed. Universal Xpert with simultaneous TPT prevented incident TB compared to either symptom screening (median 17 cases averted per 5000 patients) or Universal Xpert with delayed TPT (median 23 averted). Universal Xpert with Simultaneous TPT cost approximately $39 per incremental TPT course compared to Universal Xpert with delayed TPT. Universal Xpert testing can promote timely treatment for newly diagnosed people with HIV who have active TB. Pairing universal testing with immediate TPT will improve the promptness, uptake, and preventive effects of TPT. Simultaneous improvements to TB care cascades are needed to maximize impact.


Pretreatment Vitamin D Deficiency Is Associated With Impaired Progression-Free and Overall Survival in Hodgkin Lymphoma.
Vitamin D deficiency is described as a modifiable risk factor for the incidence of and mortality in many common cancers; however, data in Hodgkin lymphoma (HL) are lacking. We thus performed a study measuring pretreatment vitamin D levels in prospectively treated patients with HL and correlated this with clinical outcomes. A total of 351 patients from the German Hodgkin Study Group clinical trials (HD7, HD8, and HD9) were included. Fifty percent of patients were vitamin D deficient (< 30 nmol/L) before planned chemotherapy. Pretreatment vitamin D deficiency was more common in relapsed/refractory patients than matched relapse-free controls (median baseline vitamin D, 21.4 nmol/L v 35.5 nmol/L; proportion with vitamin D deficiency, 68% v 41%; P < .001). Vitamin D-deficient patients had impaired progression-free survival (10-year difference, 17.6%; 95% CI, 6.9% to 28.4%; hazard ratio, 2.13; 95% CI, 1.84 to 2.48; P < .001) and overall survival (10-year difference, 11.1%; 95% CI, 2.1% to 20.2%; hazard ratio, 1.82; 95% CI, 1.53 to 2.15; P < .001), consistent across trials and treatment groups. We demonstrated that vitamin D status is an independent predictor of outcome and hypothesized that vitamin D status might be important for the chemosensitivity of HL. We subsequently performed experiments supplementing physiologic doses of vitamin D (calcitriol) to cultured HL cell lines and demonstrated increased antiproliferative effects in combination with chemotherapy. In an HL-xenograft animal model, we showed that supplemental vitamin D (dietary supplement, cholecalciferol) improves the chemosensitivity of tumors by reducing the rate of tumor growth compared with vitamin D or chemotherapy alone. On the basis of our clinical and preclinical findings, we encourage that vitamin D screening and replacement be incorporated into future randomized clinical trials to properly clarify the role of vitamin D replacement therapy in HL.


Genome search in celiac disease.
Celiac disease (CD), a malabsorption disorder of the small intestine, results from ingestion of gluten. The HLA risk factors involved in CD are well known but do not explain the entire genetic susceptibility. To determine the localization of other genetic risk factors, a systematic screening of the genome has been undertaken. The typing information of 281 markers on 110 affected sib pairs and their parents was used to test linkage. Systematic linkage analysis was first performed on 39 pairs in which both sibs had a symptomatic form of CD. Replication of the regions of interest was then carried out on 71 pairs in which one sib had a symptomatic form and the other a silent form of CD. In addition to the HLA loci, our study suggests that a risk factor in 5qter is involved in both forms of CD (symptomatic and silent). Furthermore, a factor on 11qter possibly differentiates the two forms. In contrast, none of the regions recently published was confirmed by the present screening.


Assessment of retinoblastoma RNA reflux after intravitreal injection of melphalan.
Intravitreal injection of chemotherapy in retinoblastoma eyes with vitreous seeds may lead to a risk of extraocular tumour dissemination that has not been assessed so far. To develop a sensitive and clinically feasible technique to assess for potential retinoblastoma cell reflux after intravitreal injection of melphalan. Filter papers were cut in 6 mm diameter circles and sterilised before use. Eyes with retinoblastoma vitreous seeds (group D, International Classification) received weekly intravitreal melphalan injections (20 µg or 30 µg/dose) followed by cryotherapy as part of local treatment. Immediately after finishing the injection and cryotherapy, filter papers were placed on the injection site and on the cryoprobe tip to assess for the expression of the cone-rod homeobox gene (CRX) by real-time qPCR as a surrogate of retinoblastoma RNA. The assay was developed and validated to determine sensitivity, linearity, recovery, repeatability and reproducibility. The assay for quantitation of CRX expression was linear in the range of 1 to 1000 cells. The lowest limit of detection was one retinoblastoma cell and allowed to recover 100% of the cell load in external supplementation. A total of 14 eyes received 22 cycles of intravitreal melphalan and were evaluated for potential extraocular tumour cell dissemination using the developed technique. None of the cycles were positive for CRX in samples from the scar or from the cryoprobe tip. A sensitive and simple method of tumour cell assessment has been developed that can be used in the clinics to assess for potential extraocular dissemination after intravitreal injections to assure its performance.


Minimally invasive hysterectomy for stage IA cervical carcinoma: a survival analysis of the National Cancer Database.
To evaluate the outcomes of minimally invasive surgery for patients with stage IA cervical carcinoma undergoing hysterectomy. Patients with pathological stage IA (IA1, IA2, IA not otherwise specified) squamous, adenocarcinoma, adenosquamous carcinoma of the cervix, no history of another tumor, who underwent radical or simple hysterectomy with known mode of surgery, diagnosed between 2010 and 2015 with at least 1 month of follow-up, were drawn from the National Cancer Database. Comparisons of demographic and clinicopathologic characteristics were made with the χ2 test. The impact of minimally invasive surgery (robotic-assisted or traditional laparoscopic) on overall survival was assessed with the log-rank test following generation of Kaplan-Meier curves. A Cox model was constructed to control for confounders. A total of 1930 patients were identified; the majority (73.3%, 1414 patients) had stage IA1 disease, while 458 (23.7%) patients had stage IA2, and 58 (3%) patients had stage IA not otherwise specified. In the present cohort, 685 patients (35.5%) had open, 438 patients (22.7%) had laparoscopic, and 807 patients (41.8%) had robotic-assisted laparoscopic hysterectomy. Patients who had an open approach were more likely to undergo lymphadenectomy (58.1% vs 52.7%, p=0.021) and have radical hysterectomy (42% vs 32.4%, p<0.001). Patients who had minimally invasive surgery had a shorter hospital stay (median 1 vs 3 days, p<0.001). There was no difference in overall survival between patients who had open and minimally invasive hysterectomy (p=0.87); 4-year overall survival rates were 97.7% and 98.6%, respectively. There was no difference in overall survival between the open and minimally invasive surgery groups for patients who had simple (p=0.61; 4-year overall survival rates 97.6% and 98.7%, respectively) or radical hysterectomy (p=0.70; 4-year overall survival rates 97.8% and 98.4%, respectively). After controlling for patient age, tumor histology, and presence of lymphovascular invasion, minimally invasive hysterectomy was not associated with worse survival (HR 0.94, 95% CI 0.49 to 1.81). In a sensitivity analysis, based on 3048 patients with clinical stage IA after controlling for confounders, minimally invasive surgery was not associated with worse survival than laparotomy (HR 1.06, 95% CI 0.65 to 1.72). In a large cohort of patients with stage IA cervical carcinoma, performance of minimally invasive hysterectomy was not associated with a detrimental effect on overall survival.


Injury to the acetabular triradiate physeal cartilage.
Traumatic disruption of the acetabular triradiate physeal cartilage is an infrequent injury. When it occurs during adolescence, subsequent growth changes in acetabular morphology and congruency of the hip joint are unlikely. However, in younger children, especially those who are less than ten years old, acetabular growth abnormality is a frequent complication of this injury and may result in a shallow acetabulum similar to that seen in patients with congenital disease of the hip. By the time of skeletal maturity, disparate growth increases the incongruency of the hip joint and may lead to progressively more severe subluxation of the hip. Acetabular reconstruction may be necessary to correct the gradual subluxation of the femoral head. Variable irregularities of growth at the proximal end of the femur also may occur. In this series, nine patients with triradiate physeal-cartilage injury were classified according to the degree of displacement and the probable type of growth-plate disruption. Two main patterns of injury occurred. The first was a shearing type-1 or 2 growth-mechanism injury, with central displacement of the distal portion of the acetabulum. This injury pattern seems to have a favorable prognosis for continued normal acetabular growth, although premature closure of the triradiate physes may occur. The other pattern appeared to be a crushing type-5 growth-mechanism injury. This type has a poor prognosis, with premature closure of the triradiate physes occurring secondary to the formation of a medial osseous bridge. In either pattern, the prognosis is dependent on the age of the patient at the time of injury and on the extent of chondro-osseous disruption.


Adding realism to simulated sensors and actuators.
In this paper, we propose a statistical theoretical framework for incorporation of sensor and actuator faults in dynamic simulations of wastewater treatment operation. Sensor and actuator faults and failures are often neglected in simulations for control strategy development and testing, although it is well known that they represent a significant obstacle for realising control at full-scale facilities. The framework for incorporating faults and failures is based on Markov chains and displays the appealing property of easy transition of sensor and actuator history into a model for fault generation. The paper briefly describes Markov theory and how this is used together with models for sensor and actuator dynamics to achieve a realistic simulation of measurements and actuators.


Alternative approaches to managing respiratory tract infections: a survey of public perceptions.
Respiratory tract infections (RTIs) are a common reason for people to consult in primary care, and contribute to antibiotic overuse and antimicrobial resistance (AMR). Alternative approaches to supporting patients with RTIs may help, but it is important to understand public perceptions about these approaches before they are widely implemented. To describe public perceptions regarding finger-prick testing, back-up antibiotic prescriptions (BUPs), and alternatives to traditional consultations for RTIs, and identify factors associated with favouring these approaches. Online national survey (HealthWise Wales) with linked primary care health record data. Survey item response distributions were described. Associations between responses about consultation alternatives, BUP, and finger-prick point-of-care testing (POCT), and potential explanatory variables, were explored using logistic regression. A total of 8752 participants completed the survey between 2016 and 2018. The survey found 76.7% (n = 3807/4,966) and 71.2% (n = 3529/4,953) of responders with valid responses were in favour of being able to consult with a pharmacist or nurse in their GP surgery, or with a community pharmacist, respectively. It also showed 92.8% (n = 8034/8659) of responders indicated they would be happy to have a finger-prick test to guide antibiotic prescribing, and 31.8% (n = 2746/8646) indicated they would like to be given a BUP if their clinician thought immediate antibiotics were not required. In addition, 47.4% (n = 2342/4944) and 42.3% (n = 2095/4949) were in favour of having video and email consultations, respectively. Characteristics associated with different response options were identified. Consulting with pharmacists, using electronic communication tools, and finger-prick testing are widely acceptable approaches. BUP was described as acceptable less often, and is likely to require greater information and support when used.


The role of epidermal growth factor (EGF) and its receptor in mucosal protection, adaptation to injury, and ulcer healing: involvement of EGF-R signal transduction pathways.
Growth factors and their receptors are known to play important roles in normal cell proliferation, morphogenesis, tissue repair, and ulcer healing. Epidermal growth factor (EGF) inhibits acid secretion, exerts a trophic effect on gastroduodenal mucosa, protects gastric mucosa against injury, mediates mucosal adaptation, and accelerates gastroduodenal ulcer healing by stimulating cell migration and proliferation. EGF exerts its actions by binding to its receptor, EGF-R, a transmembrane protein tyrosine kinase, which triggers receptor dimerization, autophosphorylation, and recruitment of kinase substrates. These events result in Ras (GTP-binding protein) activation of the Ras/Raf/MAP kinase pathway, leading to phosphorylation of regulatory proteins and transcription factors and culminating in cell proliferation. Other pathways potentially activated by EGF include the phosphatidylinositol pathway and the JAK/STAT signaling pathway. Recent studies demonstrated that EGF-R-associated tyrosine kinase plays an essential role in regulating gastric mucosal cell proliferation after acute injury and further demonstrated activation of the EGF-R gene, EGF-R phosphorylation, and increased MAP kinase activity during early stages of experimental gastric ulcer healing. Finally, experimental data indicate that Helicobacter pylori vacuolating cytotoxin inhibits healing of experimental gastric ulcers, cell proliferation, binding of EGF to its receptor, EGF-induced EGF-R phosphorylation, and MAP kinase (ERK-2) activation. These H. pylori actions can explain its interference with the ulcer healing process.


Injection of air into the headspace improves fermentation of phosphoric acid pretreated sugarcane bagasse by Escherichia coli MM170.
Microaeration (injecting air into the headspace) improved the fermentation of hemicellulose hydrolysates obtained from the phosphoric acid pretreatment of sugarcane bagasse at 170°C for 10 min. In addition, with 10% slurries of phosphoric acid pretreated bagasse (180°C, 10 min), air injection into the headspace promoted xylose utilization and increased ethanol yields from 0.16 to 0.20 g ethanol/g bagasse dry weight using a liquefaction plus simultaneous saccharification and co-fermentation process (L+SScF). This process was scaled up to 80 L using slurries of acid pretreated bagasse (96 h incubation; 0.6L of air/min into the headspace) with ethanol yields of 312-347 L (82-92 gal) per tone (dry matter), corresponding to 0.25 and 0.27 g/g bagasse (dry weight). Injection of small amounts of air into the headspace may provide a convenient alternative to subsurface sparging that avoids problems of foaming, sparger hygiene, flotation of particulates, and phase separation.


Chloroquine inhibits autophagic flux by decreasing autophagosome-lysosome fusion.
Macroautophagy/autophagy is a conserved transport pathway where targeted structures are sequestered by phagophores, which mature into autophagosomes, and then delivered into lysosomes for degradation. Autophagy is involved in the pathophysiology of numerous diseases and its modulation is beneficial for the outcome of numerous specific diseases. Several lysosomal inhibitors such as bafilomycin A1 (BafA1), protease inhibitors and chloroquine (CQ), have been used interchangeably to block autophagy in in vitro experiments assuming that they all primarily block lysosomal degradation. Among them, only CQ and its derivate hydroxychloroquine (HCQ) are FDA-approved drugs and are thus currently the principal compounds used in clinical trials aimed to treat tumors through autophagy inhibition. However, the precise mechanism of how CQ blocks autophagy remains to be firmly demonstrated. In this study, we focus on how CQ inhibits autophagy and directly compare its effects to those of BafA1. We show that CQ mainly inhibits autophagy by impairing autophagosome fusion with lysosomes rather than by affecting the acidity and/or degradative activity of this organelle. Furthermore, CQ induces an autophagy-independent severe disorganization of the Golgi and endo-lysosomal systems, which might contribute to the fusion impairment. Strikingly, HCQ-treated mice also show a Golgi disorganization in kidney and intestinal tissues. Altogether, our data reveal that CQ and HCQ are not bona fide surrogates for other types of late stage lysosomal inhibitors for in vivo experiments. Moreover, the multiple cellular alterations caused by CQ and HCQ call for caution when interpreting results obtained by blocking autophagy with this drug.


Anomalously enhanced hydration of aqueous electrolyte solution in hydrophobic carbon nanotubes to maintain stability.
An understanding of the structure and behavior of electrolyte solutions in nanoenvironements is crucial not only for a wide variety of applications, but also for the development of physical, chemical, and biological processes. We demonstrate the structure and stability of electrolyte in carbon nanotubes using hybrid reverse Monte Carlo simulations of X-ray diffraction patterns. Hydrogen bonds between water are adequately formed in carbon nanotubes, although some hydrogen bonds are restricted by the interfaces of carbon nanotubes. The hydrogen bonding network of water in electrolyte in the carbon nanotubes is further weakened. On the other hand, formation of the ion hydration shell is significantly enhanced in the electrolyte in the carbon nanotubes in comparison to ion hydration in bulk electrolyte. The significant hydrogen bond and hydration shell formation are a result of gaining stability in the hydrophobic nanoenvironment.


Helenalin-mediated post-transcriptional regulation of p21(Cip1) inhibits 3T3-L1 preadipocyte proliferation.
We have previously shown that post-transcriptional mechanisms involving the 26S proteasome regulate the cyclin-dependent kinase inhibitors (CKIs), p21(Cip1) and p27(Kip1) during preadipocyte proliferation. Earlier studies further demonstrated that the anti-inflammatory, anti-carcinogenic phytochemical, helenalin is a potent inhibitor of periodic Skp2 accumulation, an F-box protein mediating SCF E3 ligase ubiquitylation and degradation of both CKIs during S phase progression. Data presented here demonstrate that helenalin dose-dependently induced G1 arrest of synchronously replicating 3T3-L1 preadipocytes. This effect occurred in the absence of discernable indices of cell toxicity or apoptosis under the conditions used in this study. Our results demonstrate that helenalin markedly increased p21 protein accumulation in both density-arrested and proliferating preadipocytes in a dose-dependent manner. This increase in p21 protein abundance occurred without change in mRNA transcript demonstrating that post-transcriptional mechanisms were involved. This notion was further supported by the modest accumulation of polyubiquitylated p21 following treatment with helenalin suggesting that suppression of targeted p21 proteolysis by the 26S proteasome contributed to helenalin-mediated p21 accumulation. The increase in p21 protein was compartmentalized to the nucleus where p21 is known to inhibit cell cycle progression. Finally, helenalin increased protein-protein interactions between p21 and cyclin-dependent kinase 2 (Cdk2) which may account in part for the anti-proliferative effect in 3T3-L1 preadipocytes.


CNQX and AMPA inhibit electrical synaptic transmission: a potential interaction between electrical and glutamatergic synapses.
Electrical synapses play an important role in signaling between neurons and the synaptic connections between many neurons possess both electrical and chemical components. Although modulation of electrical synapses is frequently observed, the cellular processes that mediate such changes have not been studied as thoroughly as plasticity in chemical synapses. In the leech (Hirudo sp), the competitive AMPA receptor antagonist CNQX inhibited transmission at the rectifying electrical synapse of a mixed glutamatergic/electrical synaptic connection. This CNQX-mediated inhibition of the electrical synapse was blocked by concanavalin A (Con A) and dynamin inhibitory peptide (DIP), both of which are known to inhibit endocytosis of neurotransmitter receptors. CNQX-mediated inhibition was also blocked by pep2-SVKI (SVKI), a synthetic peptide that prevents internalization of AMPA-type glutamate receptor. AMPA itself also inhibited electrical synaptic transmission and this AMPA-mediated inhibition was partially blocked by Con A, DIP and SVKI. Low frequency stimulation induced long-term depression (LTD) in both the electrical and glutamatergic components of these synapses and this LTD was blocked by SVKI. GYKI 52466, a selective non-competitive antagonist of AMPA receptors, did not affect the electrical EPSP, although it did block the glutamatergic component of these synapses. CNQX did not affect non-rectifying electrical synapses in two different pairs of neurons. These results suggest an interaction between AMPA-type glutamate receptors and the gap junction proteins that mediate electrical synaptic transmission. This putative interaction between glutamate receptors and gap junction proteins represents a novel mechanism for regulating the strength of synaptic transmission.


TREEFINDER: a powerful graphical analysis environment for molecular phylogenetics.
Most analysis programs for inferring molecular phylogenies are difficult to use, in particular for researchers with little programming experience. TREEFINDER is an easy-to-use integrative platform-independent analysis environment for molecular phylogenetics. In this paper the main features of TREEFINDER (version of April 2004) are described. TREEFINDER is written in ANSI C and Java and implements powerful statistical approaches for inferring gene tree and related analyzes. In addition, it provides a user-friendly graphical interface and a phylogenetic programming language. TREEFINDER is a versatile framework for analyzing phylogenetic data across different platforms that is suited both for exploratory as well as advanced studies.


Pancreatic exocrine secretion is blocked by inhibitors of methylation.
A number of early experiments suggested a relationship between methyl group metabolism and the exocrine secretion of the pancreas. These included nutritional studies showing that ethionine, the ethyl analog of methionine which inhibits cellular methylation reactions, is a specific pancreatic toxin. Other studies indicated that protein carboxymethylation might be involved. We now show that in vivo ethionine inhibits amylase secretion from freshly isolated rat pancreatic acini, while in vitro ethionine inhibits amylase secretion from the AR42J pancreatic cell line. S-Adenosylhomocysteine (SAH) is a product inhibitor of all methyltransferase reactions involving S-adenosylmethionine (SAM), and treatments that elevate cellular levels of SAH such as inhibition of S-adenosylhomocysteine hydrolase and the in vitro addition of adenosine and homocysteine result in the inhibition of amylase secretion in both isolated pancreatic acini and AR42J cells. Measurement of SAM and SAH levels in AR42J cells shows that inhibition of secretion is more closely related to elevation of SAH levels than to a decrease in the SAM/SAH ratio. Small G-proteins are carboxymethylated on the C-terminal prenylated cysteine and inhibitors of membrane-associated prenylcysteine methyltransferase, N-acetylfarnesylcysteine, N-acetylgeranylgeranylcysteine, and farnesylthioacetic acid (FTA), block secretion in AR42J cells. N-Acetylgeranylcysteine is not an inhibitor of the methyltransferase and does not inhibit amylase secretion. FTA inhibits membrane-associated prenylcysteine methyltransferase from AR42J cells with a Ki in the 45-69 microm range. These results suggest that a methylation event is needed for pancreatic exocrine secretion which may be the reversible methylation of a G-protein involved in signal transduction or membrane trafficking.


Ovarian Tumor Domain-Containing 7B Attenuates Pathological Cardiac Hypertrophy by Inhibiting Ubiquitination and Degradation of Krüppel-Like Factor 4.
Cardiac hypertrophy (CH) is a well-established risk factor for many cardiovascular diseases and a primary cause of mortality and morbidity among older adults. Currently, no pharmacological interventions have been specifically tailored to treat CH. OTUD7B (ovarian tumor domain-containing 7B) is a member of the ovarian tumor-related protease (OTU) family that regulates many important cell signaling pathways. However, the role of OTUD7B in the development of CH is unclear. Therefore, we investigated the role of OTUD7B in CH. OTUD7B knockout mice were used to assay the role of OTUD7B in CH after transverse aortic coarctation surgery. We further assayed the specific functions of OTUD7B in isolated neonatal rat cardiomyocytes. We found that OTUD7B expression decreased in hypertrophic mice hearts and phenylephrine-stimulated neonatal rat cardiomyocytes. Furthermore, OTUD7B deficiency exacerbated transverse aortic coarctation surgery-induced myocardial hypertrophy, abnormal cardiac function, and fibrosis. In cardiac myocytes, OTUD7B knockdown promoted phenylephrine stimulation-induced myocardial hypertrophy, whereas OTUD7B overexpression had the opposite effect. An immunoprecipitation-mass spectrometry analysis showed that OTUD7B directly binds to KLF4 (Krüppel-like factor 4). Additional molecular experiments showed that OTUD7B impedes KLF4 degradation by inhibiting lysine residue at 48 site-linked ubiquitination and suppressing myocardial hypertrophy by activating the serine/threonine kinase pathway. These results demonstrate that the OTUD7B-KLF4 axis is a novel molecular target for CH treatment.


Poly(trehalose): sugar-coated nanocomplexes promote stabilization and effective polyplex-mediated siRNA delivery.
When nanoparticles interact with their environment, the nature of that interaction is governed largely by the properties of its outermost surface layer. Here, we exploit the exceptional properties of a common disaccharide, trehalose, which is well-known for its unique biological stabilization effects. To this end, we have developed a synthetic procedure that readily affords a polymer of this disaccharide, poly(methacrylamidotrehalose) or "poly(trehalose)" and diblock copolycations containing this polymer with 51 repeat units chain extended with aminoethylmethacrylamide (AEMA) at three degrees of polymerization (n = 34, 65, and 84). Two series of experiments were conducted to study these diblock copolymers in detail and to compare their properties to two control polymers [PEG-P(AEMA) and P(AEMA)]. First, we demonstrate that the poly(trehalose) coating ensures colloidal stability of polyplexes containing siRNA in the presence of high salt concentrations and serum proteins. Poly(trehalose) retains the ability of trehalose to lower the phase transition energy associated with water freezing and can protect siRNA polyplexes during freeze-drying, allowing complete nanoparticle resuspension without loss of biological function. Second, we show that siRNA polyplexes coated with poly(trehalose) have exceptional cellular internalization into glioblastoma cells that proceeds with zero-order kinetics. Moreover, the amount of siRNA delivered by poly(trehalose) block copolycations can be controlled by the siRNA concentration in cell culture media. Using confocal microscopy we show that trehalose-coated polyplexes undergo active trafficking in cytoplasm upon internalization and significant siRNA-induced target gene down-regulation was achieved with an IC50 of 19 nM. These findings coupled with a negligible cytotoxicity suggests that poly(trehalose) has the potential to serve as an important component of therapeutic nanoparticle formulations of nucleic acids and has great promise to be extended as a new coating for other nanobased technologies and macromolecules, in particular, those related to nanomedicine applications.


Comparison of shear wave velocity measurements assessed with two different ultrasound systems in an ex-vivo tendon strain phantom.
The purpose of this study is to compare the reliability of SW velocity measurements of two different ultrasound systems and their correlation with the tangent traction modulus in a non-static tendon strain model. A bovine tendon was fixed in a custom-made stretching device. Force was applied increasing from 0 up to 18 Newton. During each strain state the tangent traction modulus was determined by the stretcher device, and SW velocity (m/s) measurements using a Siemens S3000 and a Supersonic Aixplorer US machine were done for shear modulus (kPa) calculation. A strong significant positive correlation was found between SW velocity assessed by the two ultrasound systems and the tangent traction modulus (r = 0.827-0.954, p < 0.001), yet all SW velocity-based calculations underestimated the reference tissue tangent modulus. Mean difference of SW velocities with the S3000 was 0.44 ± 0.3 m/s (p = 0.002) and with the Aixplorer 0.25 ± 0.3 m/s (p = 0.034). Mean difference of SW velocity between the two US-systems was 0.37 ± 0.3 m/s (p = 0.012). In conclusion, SW velocities are highly dependent on mechanical forces in the tendon tissue, but for controlled mechanical loads appear to yield reproducible and comparable measurements using different US systems.


Intestinal immunization with soluble bacterial antigens: the example of cholera toxoid.
The studies described are aimed at a better understanding of the intestinal immunological system and its role in protection against enteric infection. The cellular kinetics of the intestinal immune response to cholera toxoid were studied in rats and the protection afforded by toxoid immunization was studied in dogs. Memory was demonstrated in the gut immune system. Plasma cells containing IgA antitoxin appeared in large numbers in gut lamina propria when intraduodenal boosting followed either intraperitoneal priming or prolonged oral priming, intraperitoneal priming being the most efficient. Immunization by the intraperitoneal route alone produced no response in small bowel lamina propria. Lamina propria plasma cells were derived from precursors in Peyer's patches or mesenteric lymph nodes which migrated through the thoracic duct and systemic circulation before homing to the gut. Dogs were immunized parenterally with cholera toxin or toxoid and challenged orally with Vibrio cholerae. Protection correlated closely with serum antitoxin titres and was usually brief. Passive intravenous immunization with IgG antitoxin was also protective. In contrast, subcutaneous priming followed by oral boosting yielded longer protection without elevated serum antitoxin titres. Antitoxin was detected in jejunal washings only briefly after local boosting. The mechanism by which protection is prolonged is unclear but its greater duration after parenteral priming and oral boosting emphasizes the importance of stimulating the gut immune mechanism in attempts to immunize against enteric bacterial infections. The parenteral-oral squence may be an effective means of immunizing the intestine with non-replicating protein antigens.


Designer enediynes generate DNA breaks, interstrand cross-links, or both, with concomitant changes in the regulation of DNA damage responses.
The ability of the radiomimetic anticancer enediyne C-1027 to induce ataxia-telangiectasia mutated (ATM) and ATM and Rad3-related (ATR)-independent damage responses was discovered to reside in its unique ability to concurrently generate robust amounts of double-strand breaks (DSBs) and interstrand cross-links (ICLs) in cellular DNA. Furthermore, a single substitution to the chromophore's benzoxazolinate moiety shifted DNA damage to primarily ICLs and an ATR- but not ATM-dependent damage response. In contrast, single substitutions of the chromophore's beta-amino acid component shifted DNA damage to primarily DSBs, consistent with its induction of conventional ATM-dependent damage responses of the type generated by ionizing radiation and other radiomimetics. Thus, phosphatidylinositol 3-kinase-like protein kinase regulation of DNA damage responses is dictated by the relative proportions of DSBs and ICLs.


Comparison of efficiency and preference of metal and plastic spacers in preschool children.
The metal NebuChamber valved holding chamber (VHC) has gained wide acceptance among children with asthma. Due to its nonelectrostatic properties and larger volume, the 250-mL, metal NebuChamber delivers a greater mass of aerosol to a filter at the mouth compared with the commonly used 150-mL polypropylene AeroChamber VHC. Such in vitro results have been used to suggest that this may provide increased efficacy with the NebuChamber. No comparative efficacy data exist for preschool children with asthma. To compare efficiency and preference of metal and plastic spacers in preschool children. Children with mild-to-moderate persistent asthma received 200 microg of budesonide twice daily by NebuChamber or AeroChamber, both with the mask provided in a randomized, 2-month, crossover trial. Symptom diary cards, beta-agonist use, and preference by children and parents were compared. Thirty children (mean +/- SD age, 4.3 +/- 0.3 years) completed the study. There was no difference between the AeroChamber and NebuChamber in clinical efficacy outcomes. There was no difference between the AeroChamber and NebuChamber in parents' view of ease of use, design, acceptability by the children, and overall satisfaction. Despite a greater total dose delivered to the mouth, the NebuChamber appears no more effective than the AeroChamber and it is not preferred by patients or parents. More parents chose to continue to use the NebuChamber after the study.