The effects of a winter cover crop on Diabrotica virgifera (Coleoptera: Chrysomelidae) populations and beneficial arthropod communities in no-till maize.
The effects of an autumn-planted, spring-killed, grass cover crop (Elymus trachycaulus [Link] Gould ex Shinners) on populations of Diabrotica virgifera virgifera LeConte and its predator community were evaluated in South Dakota maize fields over two seasons. Abundance and size of D. virgifera larvae and adults and sex ratio of adults were measured in maize produced under two treatments (i.e., a winter cover crop or bare soil), as were maize root damage and the abundance and diversity of the predator communities collected on the soil surface and in the soil column. First and second instars and adults of D. virgifera were similarly abundant in the two treatments, but third instars were significantly fewer in maize planted after a winter cover crop. Larvae developed at different rates in the two treatments, and second instars were significantly smaller (head capsule width and body length) in the maize planted after a cover crop. First and third instars and adults were of similar size in the two treatments, and adult sex ratios were also similar. Although initially similar, predator populations increased steadily in the cover-cropped maize, which led to a significantly greater predator population by the time D. virgifera pupated. There was significantly less root damage in the cover-cropped maize. Predator communities were similarly diverse in both treatments. Predator abundance per plot was significantly and negatively correlated with the abundance of third instars per plot. Clearly, winter cover crops reduce D. virgifera performance and their damage to the crop, and we suspect that this reduction is caused by both environmental effects of the treatment on D. virgifera size and development, and of increased predation on the third instars of the pest. Additional data on the impact of cover crops on actual predation levels, grain yield and quality, and farmer profitability, and correlations among pest performance, crop characteristics, and predator populations and behaviors are key components of this system that remain to be addressed.


Partial and impartial ethical reasoning in health care professionals.
To determine the relationship between ethical reasoning and gender and occupation among a group of male and female nurses and doctors. Partialist and impartialist forms of ethical reasoning were defined and singled out as being central to the difference between what is known as the "care" moral orientation (Gilligan) and the "justice" orientation (Kohlberg). A structured questionnaire based on four hypothetical moral dilemmas involving combinations of (health care) professional, non-professional, life-threatening and non-life-threatening situations, was piloted and then mailed to a randomly selected sample of doctors and nurses. 400 doctors from Victoria, and 200 doctors and 400 nurses from New South Wales. 178 doctors and 122 nurses returned completed questionnaires. 115 doctors were male, 61 female; 50 nurses were male and 72 were female. It was hypothesised that there would be an association between feminine subjects and partialist reasoning and masculine subjects and impartialist reasoning. It was also hypothesised that nurses would adopt a partialist approach to reasoning and doctors an impartialist approach. No relationship between any of these variables was observed.


Body- and Movement-Oriented Interventions for Posttraumatic Stress Disorder: A Systematic Review and Meta-Analysis.
To assess the efficacy of body- and movement-oriented interventions (BMOIs) in traumatized adults with posttraumatic stress disorder (PTSD), we conducted a systematic review and meta-analysis of pertinent literature. Four bibliographical databases (PsycINFO, Ovid MEDLINE(R), EMBASE, and the Cochrane Central Register of Controlled Trials) were searched using keywords and text words for trials on BMOIs addressing PTSD. The search included articles published between October 2005 and August 2017. Studies were included if participants were adults suffering from PTSD, if BMOIs were the therapeutic strategy under investigation, and if a psychometrically evaluated standardized outcome measure for PTSD was used. No limitations for control conditions were applied. Hedges' g was computed as the effect size (ES) for the treatment versus control condition. The meta-analysis included 15 studies, which resulted in a mean ES of g = 0.85, 95% CI [0.31, 1.39], with very high heterogeneity, I2 = 91%. After removing one study as outlier, a mean effect size of g = 0.56, 95% CI [0.29, 0.82] (i.e., medium effect), still with considerable heterogeneity, I2 = 57%, was found. BMOIs seem to be effective in reducing symptoms of PTSD, but more research is needed to identify working mechanisms and to determine which types of intervention are most effective for various subgroups of patients.


Recombinant lignin peroxidase-catalyzed decolorization of melanin using in-situ generated H2O2 for application in whitening cosmetics.
Lignin peroxidase has high potential as ingredient in skin whitening cosmetics due to its high redox potential to oxidize recalcitrant melanin. Currently crude mixtures of lignin peroxidase from fungal fermentation are usually applied to cosmetics due to the intrinsic difficulties of expression and purification. However, the present study focused on heterologous expression and purification of lignin peroxidase isozyme H8 (LiPH8) from Phanerochaete chrysosporium and was further used for melanin decolorization. Results revealed that the optimum pH for melanin decolorization using LiPH8 was obtained at pH 4.0. The intermittent feeding of hydrogen peroxide (H2O2) was effectively elevating melanin decolorization efficiency up to 73%, since excessive H2O2 inactivated LiPH8. For cosmetic application, intermittent feeding of H2O2 is not feasible, thus glucose oxidase (GOx) from Aspergillus niger was employed for in-situ generation of H2O2. By optimizing the GOx and glucose concentrations, a melanin decolorization efficiency up to 63.3 ± 2.4% was obtained within 1 h and continued to 84.0 ± 1.8% in 8 h. Conclusively, lignin peroxidase-catalyzed decolorization of melanin with in-situ generated H2O2 revealed a promising approach for whitening cosmetics applications.


Insulinoma. Experience in surgical treatment.
Sixty cases of insulinoma were treated surgically. The possible causes for failure to find the tumor during exploratory surgery are discussed. Simple enucleation is preferred in most cases of single or a few insulinomas. A technique of enucleation with minimal chance of injury to adjacent pancreatic ducts and larger blood vessels can be used. When no tumor is found after thorough exploratory procedures, controlled staged resection of the pancreas under blood glucose monitoring is the procedure of choice. Postoperative complications are common but the late results are gratifying provided the operation is carried out early in the course of the disease.


Biodegradable Shape Memory Polymers in Medicine.
Shape memory materials have emerged as an important class of materials in medicine due to their ability to change shape in response to a specific stimulus, enabling the simplification of medical procedures, use of minimally invasive techniques, and access to new treatment modalities. Shape memory polymers, in particular, are well suited for such applications given their excellent shape memory performance, tunable materials properties, minimal toxicity, and potential for biodegradation and resorption. This review provides an overview of biodegradable shape memory polymers that have been used in medical applications. The majority of biodegradable shape memory polymers are based on thermally responsive polyesters or polymers that contain hydrolyzable ester linkages. These materials have been targeted for use in applications pertaining to embolization, drug delivery, stents, tissue engineering, and wound closure. The development of biodegradable shape memory polymers with unique properties or responsiveness to novel stimuli has the potential to facilitate the optimization and development of new medical applications.


Central patterning and reflex control of antennular flicking in the hermit crab Pagurus alaskensis (Benedict).
1. The effects of altering sensory input on the motoneuronal activity underlying antennular flicking have been tested. 2. Removal of the short segments of the outer flagellum results in a reduction of the number of spikes/burst in the fast flexor motoneurones A31F and A32F. 3. During a flick the delay between the burst in motoneurone A31F and the burst in motoneurone A32F is insensitive to alteration of sensory input. 4. Sensory feedback from the flexion phase of a flick is necessary for the activation of either extensor motoneurone. Evidence is presented to suggest that this feedback is primarily from joint-movement receptors at the MS-DS and DS-OF joints. 5. The results are incorporated into a model in which the patterns of flexor activity result from some specified properties of three components: a trigger system, a follower system, and the spike initiating zone of the flexor motoneurones. The trigger system determines when a flick will occur. The follower system determines the number of flexor spikes during a flick. Properties of the spike initiating zone determine the spike frequency and the timing between bursts in the flexor motoneurones. Extensor activity in the model is reflexively elicited by feedback from phasic, unidirectional receptors sensitive to joint flexion. 6. The functional significance of reflex control of extensor activity is discussed in relation to the form and proposed function of antennular flicking. It is suggested that this form of control is adapted to the function of antennular flicking because flexion at the MS-DS joint is not always necessary for the fulfilment of the fuction of a flick.


Renal transplant patients at high risk of acute rejection benefit from adequate exposure to mycophenolic acid.
To better define subpopulations in which achieving adequate mycophenolic acid (MPA) concentrations quickly would be important, a post hoc exploratory analysis on the fixed-dose concentration-controlled database was performed, comparing high- versus low-risk renal transplant patients. Renal transplant patients were treated with mycophenolate mofetil, corticosteroids, and cyclosporine A or tacrolimus. Patients were defined as "high risk" if they had one or more of the following characteristics: delayed graft function, second or third transplantation, panel reactive antibodies >15%, four or more human leukocyte antigen mismatches, or were of black race. A total of 549 patients (61%) were classified as high risk, of whom 284 were on cyclosporine A treatment and 265 on tacrolimus. In high-risk patients, the difference in rejection incidence was 14.3% in the MPA-area under the concentration (AUC) less than 30 mg hr/L vs. 7.8% in the MPA-AUC more than or equal to 30 mg hr/L groups (P=0.025) during the first month after transplantation; whereas, in low-risk patients, there were similar rejection rates (5.7% vs. 4.5%). In the subgroup of high-risk tacrolimus-treated patients, the difference in acute rejection incidence in the first month between patients with MPA-AUC0-12 less than or more than or equal to 30 mg hr/L was most pronounced: 16 of 67 patients (23.9%) vs. 18 of 173 patients (10.4%); P=0.012. The incidence of acute rejection is higher in high-risk patients if MPA-AUC0-12 is below 30 mg hr/L. In contrast, a difference in acute rejection incidence in low-risk patients with MPA-AUC0-12 less than or more than or equal to 30 mg hr/L was not observed. This supports the use of a higher mycophenolate mofetil starting dose in selected patient populations early after transplantation.


Developmentally regulated expression of insulin-like growth factors by differentiated murine teratocarcinomas and extraembryonic mesoderm.
The expression of plasma membrane receptors for insulin-like growth factors (IGFs) by PC13 embryonal carcinoma (EC) cells, and their immediate differentiated progeny PC13END was examined by binding radiolabelled IGF-I to cell monolayers. Both cell types express high-affinity IGF receptors, but the apparent number of unoccupied receptors sites falls by about 60% upon differentiation. Crosslinking studies reveal that both type 1 and type 2 IGF receptors are expressed by PC13EC cells. PC13END-cell-conditioned medium contains developmentally regulated, separable activities, one of which reacts directly with IGF-II, and the other with IGF for plasma membrane receptors. The former activity represents a soluble secreted IGF-binding protein. The latter activity is structurally and functionally similar to rat IGF-II. Polyclonal antibodies raised against purified rat IGF-II specifically recognize multiple forms of IGF in radiolabelled culture supernatants and material which closely resembles the soluble IGF-binding protein. Immunoprecipitation of radiolabelled culture supernatants with anti-rat IGF-II reveals that the differentiation of PC13EC cells is accompanied by the coexpression of IGF-like molecules and the soluble binding protein, and that IGF-like molecules are expressed by extraembryonic tissues of mesodermal origin in the early postimplantation mouse embryo. These findings show that IGF-like molecules are expressed in early mammalian development and may act in an autocrine fashion in vivo.


Review of the scientific evidence used for establishing US policies on added sugars.
The 2015 Dietary Guidelines for Americans Advisory Committee has set recommendations to limit added sugars. This action was based on the association between dietary pattern quality scores and chronic disease risk, the results of meta-analyses conducted for the World Health Organization, and data from modeling of dietary patterns for establishing the US Department of Agriculture's Healthy US-Style Eating Patterns. Recommendations provided by the 2015-2020 Dietary Guidelines for Americans were used by the US Food and Drug Administration to establish, for the first time, the mandatory declaration of added sugars and a Daily Value of added sugars for the Nutrition Facts label. This review provides an overview of the scientific evidence considered by the World Health Organization, the 2015-2020 Dietary Guidelines for Americans, and the US Food and Drug Administration for setting recent polices and regulations on added sugars and highlights important issues and inconsistencies in the evaluations and interpretations of the evidence.


Ramucirumab plus docetaxel versus placebo plus docetaxel for second-line treatment of stage IV non-small-cell lung cancer after disease progression on platinum-based therapy (REVEL): a multicentre, double-blind, randomised phase 3 trial.
Ramucirumab is a human IgG1 monoclonal antibody that targets the extracellular domain of VEGFR-2. We aimed to assess efficacy and safety of treatment with docetaxel plus ramucirumab or placebo as second-line treatment for patients with stage IV non-small-cell-lung cancer (NSCLC) after platinum-based therapy. In this multicentre, double-blind, randomised phase 3 trial (REVEL), we enrolled patients with squamous or non-squamous NSCLC who had progressed during or after a first-line platinum-based chemotherapy regimen. Patients were randomly allocated (1:1) with a centralised, interactive voice-response system (stratified by sex, region, performance status, and previous maintenance therapy [yes vs no]) to receive docetaxel 75 mg/m(2) and either ramucirumab (10 mg/kg) or placebo on day 1 of a 21 day cycle until disease progression, unacceptable toxicity, withdrawal, or death. The primary endpoint was overall survival in all patients allocated to treatment. We assessed adverse events according to treatment received. This study is registered with ClinicalTrials.gov, number NCT01168973. Between Dec 3, 2010, and Jan 24, 2013, we screened 1825 patients, of whom 1253 patients were randomly allocated to treatment. Median overall survival was 10·5 months (IQR 5·1-21·2) for 628 patients allocated ramucirumab plus docetaxel and 9·1 months (4·2-18·0) for 625 patients who received placebo plus docetaxel (hazard ratio 0·86, 95% CI 0·75-0·98; p=0·023). Median progression-free survival was 4·5 months (IQR 2·3-8·3) for the ramucirumab group compared with 3·0 months (1·4-6·9) for the control group (0·76, 0·68-0·86; p<0·0001). We noted treatment-emergent adverse events in 613 (98%) of 627 patients in the ramucirumab safety population and 594 (95%) of 618 patients in the control safety population. The most common grade 3 or worse adverse events were neutropenia (306 patients [49%] in the ramucirumab group vs 246 [40%] in the control group), febrile neutropenia (100 [16%] vs 62 [10%]), fatigue (88 [14%] vs 65 [10%]), leucopenia (86 [14%] vs 77 [12%]), and hypertension (35 [6%] vs 13 [2%]). The numbers of deaths from adverse events (31 [5%] vs 35 [6%]) and grade 3 or worse pulmonary haemorrhage (eight [1%] vs eight [1%]) did not differ between groups. Toxicities were manageable with appropriate dose reductions and supportive care. Ramucirumab plus docetaxel improves survival as second-line treatment of patients with stage IV NSCLC. Eli Lilly.


Stereochemical analysis of deuterated alkyl chains by MS/MS
Vicinally deuterated sec-alkyl phenyl ethers, CH(3)(CH(2))(m)()CH(OPh)CHD(CH(2))(n)()CH(3), display significant differences in mass spectra between threo and erythro stereoisomers. MS/MS experiments, in which parent ions of a single mass are selected and their fragmentation patterns subsequently measured, show that alkene expulsion represents virtually the only decomposition pathway. Two types of MS/MS experiment are reported: mass-analyzed ion kinetic energy (MIKE) spectroscopy of metastable ions and collisionally activated decomposition (CAD) of stable ions. The expulsion of a deuterated alkene from a monodeuterated precursor yields ionized phenol, PhOH(*)()(+) (m/z 94). The expulsion of an undeuterated alkene yields PhOD(*)()(+) (m/z 95). Without exception, the ratios (PhOD(*)()(+)/PhOH(*)()(+)) from precursors in the threo series have values greater than their diastereomers in the erythro series. The ratio of ratios, r = PhOD(*)()(+)/PhOH(*)()(+) for the threo divided by PhOD(*)()(+)/PhOH(*)()(+) for the erythro, has a value of 1.2 for the 2-phenoxy-3-deuteriobutanes and larger values for all of the higher homologues up through the monodeuterated phenoxyoctanes (m + n = 4). The highest degree of stereoselectivity, r = 5.8, is measured for 3-phenoxy-4-deuteriohexane. Experiments with multiply deuterated analogues show that alkene elimination is highly regioselective, unlike the corresponding decompositions of ionized sec-alcohols or their acetates. The fact that a large fraction of ionized sec-alkyl phenyl ethers undergo stereospecific syn-elimination means that mass spectrometry has a useful capacity to distinguish one isotopically labeled diastereomer from another.


Continuum of reflex excitability in hemiplegia: influence of muscle length and muscular transformation after heel-cord lengthening and immobilization on the pathophysiology of spasticity and clonus.
The electromyographic (EMG) responses to tendon taps at the ankle and ensuing muscular twitch forces and temporal parameters were studied at varying angles across the joint range in 18 children, aged 3 to 14.9 years, with congenital hemiparetic cerebral palsy and 22 healthy (control) children, aged 3 to 13.6 years. Those subjects with hemiparesis were community ambulators without assistance. In all subjects, passive muscle stretch caused a waxing of the reflex EMG and twitch force near neutral (with the sole of the foot at right angles to the tibia) and a diminution of these with further dorsiflexion. Twitch times increased with each dorsiflexing increment, being slowest at maximum dorsiflexion and fastest at the resting plantarflexion angle. Heterogeneity of the hemiparetic-limb data is evident when compared with data of non-paretic and unaffected limbs, with clear differences in the clonic (fast twitch) as opposed to non-clonic (slow twitch) muscles. In four cases with clinical clonus, clonus frequency was reduced by passive dorsiflexion. Plaster immobilization for 1 month produced clonus which was previously absent in one subject, and caused a fast-twitch phenotype to emerge in two subjects. Follow-up after heel-cord lengthening in one subject showed that clonus frequency diminished from 9 to 3 Hz with slowing and strengthening of muscle-twitch phenotype. Short- and long-term peripheral manipulations appear to regulate neuromuscular excitability according to whether muscles are loaded or unloaded. Although damage to the nervous system provides the setting for reflex excitability, the data suggest that the muscle length (which specifies the joint angle) and the muscle-twitch phenotype of any given limb for any given case appear to dictate the actual speed and strength of reflex muscle-twitch and clonus profiles. This study illustrates how peripheral manipulations of muscles and tendons may alter the expression of what have hitherto been considered as exclusively central phenomena.


How good are surgeons at identifying appendicitis? Results from a multi-centre cohort study.
Convincing arguments for either removing or leaving in-situ a macroscopically normal appendix have been made, but rely on surgeons' accurate intra-operative assessment of the appendix. This study aimed to determine the inter-rater reliability between surgeons and pathologists from a large, multicentre cohort of patients undergoing appendicectomy. The Multicentre Appendicectomy Audit recruited consecutive patients undergoing emergency appendicectomy during April and May 2012 from 95 centres. The primary endpoint was agreement between surgeon and pathologist and secondary endpoints were predictors of this disagreement. The final study included 3138 patients with a documented pathological specimen. When surgeons assessed an appendix as normal (n = 496), histopathological assessment revealed pathology in a substantial proportion (n = 138, 27.8%). Where surgeons assessed the appendix as being inflamed (n = 2642), subsequent pathological assessment revealed a normal appendix in 254 (9.6%). There was overall disagreement in 392 cases (12.5%), leading to only moderate reliability (Kappa 0.571). The grade of surgeon had no significant impact on disagreement following clinically normal appendicectomy. Females were at the highest risk of false positives and false negatives and pre-operative computed tomography was associated with increased false positives. This multi-centre study suggests that surgeons' judgements of the intra-operative macroscopic appearance of the appendix is inaccurate and does not improve with seniority and therefore supports removal at the time of surgery.


Acetylcholine receptor clustering associates with proteoglycan biosynthesis in C2 variant and heterkaryon muscle cells.
Several lines of evidence have suggested roles for proteoglycans (PGs) in acetylcholine receptor (AChR) clustering on muscle cells. One line of evidence comes from the correlation between a defect in the biosynthesis of glycosaminoglycans (GAGs), the defining carbohydrates of PGs, and the failure of spontaneous AChR clustering in the S27 cell line, a genetic variant of the C2 muscle cell line. Two approaches were used in the present study to investigate whether GAG and AChR clustering defects are causally linked. First, the formation of AChR clusters was examined in two more variant lines, S11 and S26, also isolated from the C2 muscle cell line on the basis of deficiencies in GAG biosynthesis. S11 and S26, like S27, are also defective in AChR clustering. Ion exchange analysis of the GAGs made by the S11, S26, and S27 lines revealed that the defects in GAG biosynthesis differ between the three lines. Second, heterokaryon myotubes formed between pairs of the GAG defective variants were tested for complementation in both AChR clustering and GAG biosynthesis. AChR clusters were conspicuous on individual heterokaryon myotubes, and GAG biosynthesis was restored to near wild type levels in the heterokaryon cultures. Complementation in GAG biosynthesis corroborates the biochemical data that the relevant mutations in the genetic variants are in different genes and establishes that the defects are not dominant. The consistent correlation between GAG defects and the failure of AChR clustering across three independent genetic variants and the complementary association of GAG biosynthesis with AChR clustering in heterokaryon myotubes argues against a chance association of the two phenotypes and for a causal relationship between PGs and AChR clustering. A prominent chondroitin sulfate peak correlated with AChR clustering in the heterokaryon cultures. This is consistent with earlier results suggesting that chondroitin sulfate in general is required for the spontaneous clustering of AChRs in C2 cultures and further suggests that a particular chondroitin sulfate proteoglycan may be essential for the clustering process.


In vitro efficacy of a polyhexamethylene biguanide-impregnated gauze dressing against bacteria found in veterinary patients.
To evaluate the in vitro efficacy of polyhexamethylene biguanide (PHMB)-impregnated gauze dressing in limiting the growth of bacteria both within and underneath the dressing. In vitro study. Squares of PHMB-impregnated and control gauze were placed on agar plates inoculated with 1 of 11 bacterial species, including 8 multi-resistant organisms. Growth under the gauze was assessed qualitatively after 24-hour incubation. Repeated use of sponges was used to evaluate residual inhibitory activity against Micrococcus lutea and Staphylococcus schleiferi ss. schleiferi. In a second procedure, PHMB-impregnated and control gauze squares were placed in sterile plastic wells and inoculated with 1 of 5 bacterial species, including Pseudomonas spp. and Klebsiella spp. Inhibition of bacterial growth within and underneath the dressing after 24-hour incubation was evaluated by quantifying the numbers of bacteria on the well floor and within each square. PHMB-impregnated gauze provided greater inhibition of growth of 4/4 Gram-positive species and 2/6 Gram-negative species on inoculated plates compared with control gauze. Residual inhibitory activity of PHMB-impregnated gauze was significantly greater against M. lutea on all days and against S. schleiferi ss. schleiferi on days 1 and 4 compared with control. No bacteria were recovered from inoculated PHMB-impregnated gauze squares placed in sterile wells or from the well floor underneath. More than 9 x 10(5) colony-forming units (CFU) were recovered from inoculated control samples placed in sterile wells and more than 8.4 x 10(4) CFU were recovered from control well floors. PHMB-impregnated gauze dressing, when placed on inoculated agar plates, reduces growth of underlying bacteria, particularly Gram-positive species. Wet-inoculated PHMB-impregnated dressing prevents growth of Gram-positive and Gram-negative bacteria both within and underneath the dressing. PHMB-impregnated dressings may be useful for reducing contamination of underlying wounds by bacterial pathogens.


Partial characterization of a cellular factor that regulates the double-stranded RNA-dependent eIF-2 alpha kinase in 3T3-F442A fibroblasts.
The interferon-induced double-stranded RNA-dependent eIF-2 alpha kinase (dsI) has an established role in mediating part of interferon's antiviral effects. Numerous studies have suggested that dsI also has regulatory functions in cells not infected with virus. Our previous results have indicated that the activation of this kinase may be an important regulatory signal in controlling growth arrest of mouse 3T3-F442A fibroblasts prior to their subsequent differentiation to adipocytes. Here, we report that extracts from 3T3-F442A cells cultured under conditions nonpermissive for differentiation exhibit significantly reduced dsI activity and that this reduction is due, at least in part, to the presence of elevated levels of a novel inhibitor of dsI activation (dRF). This inhibitor is also detected in reduced amounts in extracts from cells cultured under conditions which are permissive for differentiation. We have achieved a 1,000-fold purification of dRF activity, and highly purified dRF preparations were found to be greatly enriched for a 15-kDa protein that was greater than 90% pure. Our results indicate that dRF is not a protein phosphatase or protease but a reversible inhibitor of dsI autophosphorylation. In addition, our results imply that dRF is a physiologic regulator of dsI, since dRF activity correlates with the ability of 3T3-F442A cells to undergo adipose conversion.


Case for raising the minimum legal age of tobacco sale to 25.
Restricting youth access to tobacco is an essential component of a comprehensive tobacco control policy. While there has been a growing movement to raise the minimum legal age (MLA) of purchasing tobacco from 18 to 21, more restrictive measures, such as raising the MLA to 25 (MLA25), have been criticised as being overly restrictive on adult's free choice. We argue that, even within a policy approach that prioritises freedom of choice, there is a strong case for MLA25 in view of neurobiological evidence which shows that, before age 25, people are neurobiologically vulnerable to developing an addiction. We discuss further considerations for an MLA25 policy, in particular its potential impact on the free choice of young adults to start or quit smoking, potential public health impact and potential effectiveness considering that most underage youth source cigarettes from older peers.


Distribution patterns of fungal taxa and inferred functional traits reflect the non-uniform vertical stratification of soil microhabitats in a coastal pine forest.
In boreal systems, soil profiles typically consist of distinct stratified horizons, with organic layers at the surface overlying deeper mineral horizons providing microhabitat variation along a depth gradient, and vertical stratification of fungal communities along such soil profiles is commonly observed. We studied fungal community structure in a coastal pine forest along a gradient of decreasing influence from the coast. In this system, the vertical stratification pattern of soil microhabitats (defined here as organic, mineral with roots and mineral without roots: O, MR and MN, respectively) is non-uniform; organic horizons are sometimes buried under drifting sand dunes. Our results show that soil microhabitats are distinct with respect to physiochemical characteristics, community composition and OTU richness. While community composition was partly related to depth and distance from the coastal forest edge, microhabitat appeared to have the strongest influence. A closer inspection of the OTUs with the highest relative sequence abundance within each microhabitat revealed that microhabitats support functionally distinct fungal communities with respect to trophic mode and growth morphology. These results suggest that in coastal pine forests, variation in soil microhabitats contributes to the high fungal diversity found belowground and may play an important role in optimizing nutrient cycling.


Renal cysts of inv/inv mice resemble early infantile nephronophthisis.
Cystic kidney disease has been linked to mutations in the Invs gene in mice with inversion of embryonic turning (inv/inv) and the INVS (NPHP2) gene in infants with nephronophthisis type 2 (NPHP2). The inv mouse model features multiorgan defects including renal cysts, altered left-right laterality, and hepatobiliary duct malformations transmitted in an autosomal recessive manner. Affected mice usually die of renal and liver failure by postnatal day 7. Although cardiopulmonary and liver anomalies have been carefully detailed, renal cysts have yet to be fully characterized in inv/inv. By use of three-dimensional visualization by two-photon microscopy, this study provides the first comprehensive analysis of in situ cyst formation and progression in inv/inv kidneys. At embryonic day 15, there is dilatation of Bowman's capsule followed temporally by corticomedullary cysts involving collecting ducts, proximal tubules, and thick ascending limbs. Collecting ducts of newborn inv/inv mice are uniformly and diffusely cystic from medulla to cortex, with normal diameters found only at their most proximal tips. Proximal tubules form fusiform cysts that alternate with segments of normal or narrowed caliber along torturous convolutions. Because defective cilia have been linked to situs inversus and cystogenesis, we examined inv/inv cilia by scanning and transmission electron microscopy. The former detected monocilia of expected length in cystic collecting ducts and proximal tubules; the latter demonstrated the usual 9 + 2 pattern in respiratory cilia. The inv mutant mouse has renal cysts resembling infantile NPHP2 and will provide broader insight into the role cilia play in renal cystogenesis.


Prenatally diagnosed Turner syndrome and cystic hygroma: incidence and reasons for referrals.
The objective of this study was to evaluate the incidence and reasons for referrals for prenatally detected Turner syndrome and cystic hygroma cases among prenatal cases performed between 1998 and 2007. In this study 3,595 amniocentesis, chorionic villus and cordocenthesis materials obtained between 1998 and 2007 were evaluated. Among prenatal cases, 23 Turner syndrome cases were also evaluated according to their referral reasons. Among the indications of prenatal cases, cystic hygroma was evaluated according to karyotype results. Twenty-three cases were Turner Syndrome in which 7 cases were detected to have mosaic pattern. The indications for prenatal diagnosis for the cases were cystic hygroma in 11 cases, missed abortion in 6 cases, advanced maternal age in 5 cases and positive screening test results in 1 case. Among 18 cases having cystic hygroma detected by ultrasonography, 8 cases (44.4%) were found to have a 45,X karyotype, 3 cases were found to be mosaic Turner syndrome (16.7%), 5 cases (27.7%) had normal karyotype, 1 case (5.6%) 47,XX,+13 and 1 case (5.6%) 47,XX,+21. The present study indicates the importance of cystic hygroma in prenatal diagnosis of Turner Syndrome and other aneuploidies.


Endometrial receptivity and implantation are not affected by the presence of uterine intramural leiomyomas: a clinical and functional genomics analysis.
Uterine leiomyomas are the most frequent benign tumors during reproductive age. Whether intramural leiomyomas cause infertility and should be removed is controversial because no study has addressed the underlying mechanism of infertility. The objective of the study was to test the effect of intramural leiomyomas on endometrial function by comparing gene during the window of implantation and implantation in an oocyte donation program, in which the quality of the embryos replaced is similar and the endocrine environment of the endometrium is standardized by exogenous steroids. Human endometria of women with single intramural leiomyomas (group A, <5 cm and group B, > or =5 cm) and controls (group C) were collected on day LH+7 and processed for histology and gene expression analysis, using different methods and validated by quantitative RT-PCR. To compare in vitro fertilization outcome, a total of 1035 cases from our oocyte donation database were included, comprising patients with one fibroid less than 5 cm (A1, n = 532); two leiomyomas less than 5 cm (A2, n = 128); three or more leiomyomas less than 5 cm (A3, n = 125); one fibroid 5 cm or greater (B, n = 22); and two control groups: C1 (n = 93), women with previous myomectomy; and C2 (n = 135), women without uterine pathology treated on the same dates as C1. There was a strong positive and negative correlation in the expression profile of 69 genes according to the leiomyomas's size, but only three of the 25 genes related to the window of implantation were dysregulated. Term pregnancy rates after oocyte donation were 36.9, 34.1, 39.0, 36.4, 39.2, and 42.6% (P = 0.769) among the established groups. Similarly, no correlation between implantation and miscarriage with leiomyoma number and size was found. This study provides evidence that intramural leiomyomas not affecting the endometrial cavity alters the expression pattern of some endometrial genes, but the genes involved in implantation are not affected. This is confirmed by leiomyomas having no effect on oocyte donation outcome when the size and number of leiomyomas are analyzed.


Serial assessment of mortality in the neonatal intensive care unit by algorithm and intuition: certainty, uncertainty, and informed consent.
Does predictive power for outcomes of neonatal intensive care unit (NICU) patients get better with time? Or does it get worse? We determined the predictive power of Score for Neonatal Acute Physiology (SNAP) scores and clinical intuitions as a function of day of life (DOL) for newborn infants admitted to our NICU. We identified 369 infants admitted to our NICU during 1996-1997 who required mechanical ventilation. We calculated SNAP scores on DOL 1, 3, 4, 5, 7, 10, 14, 21, 28, and weekly thereafter until either death or extubation. We also asked nurses, residents, fellows, and attendings on each day of mechanical ventilation: "Do you think this child is going to live to go home to their family, or die before hospital discharge?" Two thousand twenty-eight SNAP scores were calculated for 285 infants. On DOL 1, SNAP for nonsurvivors (24 +/- 8.7 [standard deviation]) was significantly higher than SNAP for survivors (13 +/- 6.1). However, this difference diminished steadily and by DOL 10 was no longer statistically significant (12.7 +/- 4.9 vs 10.0 +/- 4.8). On each NICU day, at all ranges of SNAP scores, there were at least as many infants who would ultimately survive as would die. Consequently, the positive predictive value of any SNAP value for subsequent mortality was <0.5 on all NICU days. Prediction profiles were obtained for 230 ventilated infants reflecting over 11 000 intuitions obtained on 2867 patient days. One hundred fifty-seven (81%) of 192 survivor profiles displayed consistent accurate prediction profiles-at least 90% of their NICU ventilation days were characterized by 100% prediction of survival. Twenty-five (13%) of 192 surviving infants survived somewhat unexpectedly; that is, after at least 1 day characterized by at least 1 estimate of "death." Thirty-three (60%) of the 55 nonsurvivors died before DOL 10. Eighty-two percent of the prediction profiles for these early dying infants were homogeneous, dismal, and accurate. Twenty-two (40%) of the 55 nonsurvivors died after DOL 10. Seventeen (78%) of these 22 late-dying infants were predicted to live by many observers on many hospital days. Sixty-one (30%) of 230 profiled patients had at least 1 NICU day characterized by at least 1 prediction of death; 26/61 (43%) of these patients were incorrectly predicted; that is, they survived. Seventeen infants who were predicted to die during but survived nonetheless were assessed neurologically at 1 year. Fourteen (82%) of these 17 were not neurologically normal-8 were clearly abnormal, 1 suspicious, and 5 had died. If absolute certainty about mortality is the only criterion that can justify a decision to withhold or withdraw life-sustaining treatment in the NICU, these data would make such decisions difficult on the first day of life, and increasingly problematic thereafter. However, if we acknowledge that medicine is inevitably an inexact science and that clinical predictions can never be perfect, we can ask the more interesting question of whether good but less-than-perfect predictions of imprecise but ethically relevant clinical outcomes can still be useful. We think that they can-and that they must.


Isolation and characterization of the tubular organelles induced by fumarate reductase overproduction in Escherichia coli.
Strains of Escherichia coli amplifying the intrinsic membrane enzyme fumarate reductase accommodate the overproduced enzyme by increasing the amount of membrane material, in the form of intracellular tubular structures. These tubules have been observed in strains harbouring multicopy frd plasmids and in ampicillin hyper-resistant strains. A procedure has been developed for isolation of tubules nearly free of cytoplasmic membrane. Using protein A-gold labelling and optical diffraction of electron micrographs, a model for tubule structure is proposed. The tubules have a lower lipid/protein ratio than the cytoplasmic membrane, with the enzyme accounting for greater than 90% of the protein in the tubules. Both cytoplasmic membranes and tubules from amplified strains are enriched in cardiolipin and have a more fluid fatty acid composition than wild-type strains. Mutants defective in cardiolipin synthesis produce tubules in response to excess fumarate reductase, but these tubules have an altered appearance, indicating that lipid-protein interactions may be important for tubule assembly.


Recent advances in CE and CEC of peptides (2007-2009).
The review presents a comprehensive survey of recent developments of high-performance capillary electromigration methods, zone electrophoresis, ITP, IEF, affinity electrophoresis, EKC and electrochromatography, and their application to analysis, preparation and physicochemical characterization of peptides. New approaches to the theoretical description and experimental investigation of electromigration properties of peptides, and to methodology of their separations, such as sample preparation, adsorption suppression, EOF control and detection, are described. New developments in particular CE and CEC modes are reported and several types of their applications to peptide analysis are presented: conventional qualitative and quantitative analysis, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Few examples of micropreparative peptide separations are shown and capabilities of CE and CEC techniques to provide important physicochemical characteristics of peptides are demonstrated.


Evolving techniques in text-based medical consultation - Physicians' long-term experiences at an Ask the doctor service.
Both the demands and the options for patients to communicate with health care providers utilizing eHealth solutions are increasing. Some patients, or relatives to patients, want to consult another health care provider than the regular one, merely in text. To improve text-based medical consultation by learning from long-term experiences. Physicians with comprehensive experience of answering free-text medical inquiries at an official health portal in Sweden were interviewed. The interviews were analyzed using a grounded theory approach. Over time, the interviewed physicians developed strategies on how to formulate the answer to a medical inquiry from a previously unknown inquirer. The answering physicians experienced their primary role as providers of medical information and as mediators between an inquirer and the regular health care provider. Many of the answering physicians experienced a personal development with improved communication skills, also in face-to-face meetings with patients. Text-based medical consultation is part of an expanding area in eHealth. The development of strategies, guidelines, ethical considerations as well as educational efforts are needed to optimize the skills of asynchronous text-based health consultation.


PI3K signaling regulates rapamycin-insensitive translation initiation complex formation in vaccinia virus-infected cells.
How vaccinia virus (VV) regulates assembly of the host translation initiation complex eIF4F remains unclear. Here, we show that VV activated host PI3K to stimulate downstream mammalian target of rapamycin (mTOR), a kinase that inactivates the translational repressor 4E-BP1. However, although the mTOR inhibitor rapamycin suppressed VV-induced inactivation of 4E-BP1, it failed to inhibit eIF4F assembly. In contrast, PI3K inhibition in VV-infected cells increased the abundance of hypophosphorylated 4E-BP1 and disrupted eIF4F complex formation. PI3K signaling, therefore, plays a critical role in regulating protein production during VV infection, at least in part by controlling the abundance and activity of 4E-BP1.


Substance P and acetylcholine both suppress the same K+ current in dissociated smooth muscle cells.
The effect of substance P on freshly dissociated gastric smooth muscle cells was examined electrophysiologically. Substance P caused depolarization, associated with a membrane conductance decrease, which led to the generation of action potentials and contraction. When the membrane potential was held constant under voltage clamp, substance P induced a net inward current, also associated with a conductance decrease. The net inward current resulted from suppression of an outward K+ current, one which resembled the acetylcholine-sensitive M-current in these cells. When substance P maximally suppressed this outward K+ current, acetylcholine (ACh) had no additional effect. Conversely, when ACh fully suppressed the M-current, substance P was without additional effect. These results indicate that substance P suppresses the same outward K+ current affected by ACh. Suppression of M-current by substance P was observed in approximately half (44 of 85) of the cells studied in these experiments. In those cells that did not respond to substance P, ACh was nevertheless capable of suppressing the M-current. Thus both substance P and cholinergic agonists appear to exert their excitatory effects on smooth muscle cells by inhibiting a common K+ current.


Factors predicting mortality in elderly patients on dialysis.
Elderly patients are increasingly enrolled in dialysis programs and present a series of special characteristics due to their high morbidity and mortality. To describe the characteristics of incident dialysis patients aged >75 years, their comorbidities and their admissions, with a view to determining the factors that influence their course and mortality. The study included all patients aged >75 years who started dialysis in our center since January 2000. The mean duration of follow-up was 3.3 +/- 2.2 years. Data were collected on incident comorbidity, admissions and their causes. A total of 139 patients were included, with a mean age of 78.6 +/- 2.6 years (67.6% males, 33.8% diabetic and 7.9% on peritoneal dialysis). Three groups were established according to age: 75-79, 80-85 and >85 years. The most frequent comorbidities were chronic obstructive pulmonary disease (25.9%), ischemic heart disease (25.2%), heart failure (25.9%), neoplasms (23.7%), peripheral vascular disease (23.7%), cerebrovascular disease (18.7%) and arterial hypertension (81%). The Charlson index was calculated, not adjusted for age, and comorbidity tertiles were established. During follow-up, the patients presented 0.82 +/- 0.99 admissions/patient/year, with a duration of 12.1 +/- 20.6 days/patient/year. The main causes of admission were infection (33%), vascular access problems (27%) and peripheral vascular events (14%). A total of 61 patients died (44%), and 4 underwent kidney transplantation (2.9%). The mean duration of follow-up of the transplanted patients was 3.6 +/- 1.8 years. The main causes of death were infection (32%), cardiovascular problems (28.3%) and neoplastic disease (11.3%). The global survival rate was 90, 82 and 53% after 1, 2 and 5 years, respectively. No significant differences in survival or annual admission rate were found in relation to age group and dialysis technique. In contrast, the annual admission rate and days of admission were directly correlated to the Charlson index (p = 0.009 and p = 0.032, respectively). No significant differences in the Charlson index were found between the patients on hemodialysis and those subjected to peritoneal dialysis. In the univariate model, the factors associated to mortality were diabetes, chronic obstructive pulmonary disease, heart failure and the Charlson index. In the multivariate model, only the Charlson index remained as an independent predictive factor (p = 0.006). Unlike the general population, age does not influence mortality or admissions in elderly patients on dialysis. Incident comorbidity is the factor exerting the greatest influence upon mortality and admissions. Advanced age in itself should not be regarded as an excluding factor for starting dialysis.


[Cloning, expression and characterization of a gamma-butyrobetaine hydroxylase gene bbh from Pseudomonas sp. L-1].
Gamma-butyrobetaine hydroxylase is an enzyme that catalyzes the last step in the biosynthesis of L-carnitine. We cloned, expressed and characterized a gamma-butyrobetaine hydroxylase gene bbh from Pseudomonas sp. L-1, to facilitate the production of L-carnitine using microorganisms. We cloned bbh gene by PCR, and then cloned the open reading frame of bbh into pET-15b vector and expressed by Isopropyl beta-D-1-thiogalactopyranoside (IPTG) induction. After His-Bind Resin purification, the characteristics of BBH were studied. The three-dimensional structure of BBH monomer was modeled by SWISS-MODEL Workspace and resting cells were used for L-carnitine transformation. We cloned a gamma-butyrobetaine hydroxylase gene bbh (GenBank: JQ250036) from Pseudomonas sp. L-1 and expressed the gene in Escherichia coli BL21(DE3). BBH fusion protein was a homodimer, and the molecular weight of subunit was about 46.5kDa. The optimal temperature and pH was 30 degrees C and pH 7.5. The enzyme was stable below 45 degrees C. The enzyme was most stable at pH 6.0. We used resting cells of recombinant E. coli for L-carnitine biotransformation, after incubated at 30 degrees C and pH 7.0 for 31 h, the concentration of L-carnitine reached 12.7 mmol/L. The bbh gene from Pseudomonas sp. L-1 strain is remarkably different from that of reported one. The gamma-butyrobetaine hydroxylase expressed by this gene could effectively transform gamma-butyrobetaine for L-carnitine production. Beside by reporting of a bbh gene from bacteria, this research also provided a new process for biotransformation of L-carnitine.


Moralization in social networks and the emergence of violence during protests.
In recent years, protesters in the United States have clashed violently with police and counter-protesters on numerous occasions1-3. Despite widespread media attention, little scientific research has been devoted to understanding this rise in the number of violent protests. We propose that this phenomenon can be understood as a function of an individual's moralization of a cause and the degree to which they believe others in their social network moralize that cause. Using data from the 2015 Baltimore protests, we show that not only did the degree of moral rhetoric used on social media increase on days with violent protests but also that the hourly frequency of morally relevant tweets predicted the future counts of arrest during protests, suggesting an association between moralization and protest violence. To better understand the structure of this association, we ran a series of controlled behavioural experiments demonstrating that people are more likely to endorse a violent protest for a given issue when they moralize the issue; however, this effect is moderated by the degree to which people believe others share their values. We discuss how online social networks may contribute to inflations of protest violence.


Transvenous sonographically guided percutaneous access for treatment of an indirect carotid cavernous fistula.
When standard transarterial or transvenous interventional techniques failed to provide access to treat an indirect carotid cavernous fistula, we used sonographically guided direct percutaneous access through the facial vein to successfully embolize a lesion.


Intensively Cultivated Landscape and Varroa Mite Infestation Are Associated with Reduced Honey Bee Nutritional State.
As key pollinators, honey bees are crucial to many natural and agricultural ecosystems. An important factor in the health of honey bees is the availability of diverse floral resources. However, in many parts of the world, high-intensity agriculture could result in a reduction in honey bee forage. Previous studies have investigated how the landscape surrounding honey bee hives affects some aspects of honey bee health, but to our knowledge there have been no investigations of the effects of intensively cultivated landscapes on indicators of individual bee health such as nutritional physiology and pathogen loads. Furthermore, agricultural landscapes in different regions vary greatly in forage and land management, indicating a need for additional information on the relationship between honey bee health and landscape cultivation. Here, we add to this growing body of information by investigating differences in nutritional physiology between honey bees kept in areas of comparatively low and high cultivation in an area generally high agricultural intensity in the Midwestern United States. We focused on bees collected directly before winter, because overwintering stress poses one of the most serious problems for honey bees in temperate climates. We found that honey bees kept in areas of lower cultivation exhibited higher lipid levels than those kept in areas of high cultivation, but this effect was observed only in colonies that were free of Varroa mites. Furthermore, we found that the presence of mites was associated with lower lipid levels and higher titers of deformed wing virus (DWV), as well as a non-significant trend towards higher overwinter losses. Overall, these results show that mite infestation interacts with landscape, obscuring the effects of landscape alone and suggesting that the benefits of improved foraging landscape could be lost without adequate control of mite infestations.


Tumor cell-targeted delivery of CRISPR/Cas9 by aptamer-functionalized lipopolymer for therapeutic genome editing of VEGFA in osteosarcoma.
Osteosarcoma (OS) is a highly aggressive pediatric cancer, characterized by frequent lung metastasis and pathologic bone destruction. Vascular endothelial growth factor A (VEGFA), highly expressed in OS, not only contributes to angiogenesis within the tumor microenvironment via paracrine stimulation of vascular endothelial cells, but also acts as an autocrine survival factor for tumor cell themselves, thus making it a promising therapeutic target for OS. CRISPR/Cas9 is a versatile genome editing technology and holds tremendous promise for cancer treatment. However, a major bottleneck to achieve the therapeutic potential of the CRISPR/Cas9 is the lack of in vivo tumor-targeted delivery systems. Here, we screened an OS cell-specific aptamer (LC09) and developed a LC09-functionalized PEG-PEI-Cholesterol (PPC) lipopolymer encapsulating CRISPR/Cas9 plasmids encoding VEGFA gRNA and Cas9. Our results demonstrated that LC09 facilitated selective distribution of CRISPR/Cas9 in both orthotopic OS and lung metastasis, leading to effective VEGFA genome editing in tumor, decreased VEGFA expression and secretion, inhibited orthotopic OS malignancy and lung metastasis, as well as reduced angiogenesis and bone lesion with no detectable toxicity. The delivery system simultaneously restrained autocrine and paracrine VEGFA signaling in tumor cells and could facilitate translating CRISPR-Cas9 into clinical cancer treatment.


Substrate specificity and inducibility of TACE (tumour necrosis factor alpha-converting enzyme) revisited: the Ala-Val preference, and induced intrinsic activity.
Tumour necrosis factor alpha (TNF alpha)-converting enzyme (TACE/ADAM-17, where ADAM stands for a disintegrin and metalloproteinase) releases from the cell surface the extracellular domains of TNF and several other proteins. Previous studies have found that, while purified TACE preferentially cleaves peptides representing the processing sites in TNF and transforming growth factor alpha, the cellular enzyme nonetheless also sheds proteins with divergent cleavage sites very efficiently. More recent work, identifying the cleavage site in the p75 TNF receptor, quantifying the susceptibility of additional peptides to cleavage by TACE and identifying additional protein substrates, underlines the complexity of TACE-substrate interactions. In addition to substrate specificity, the mechanism underlying the increased rate of shedding caused by agents that activate cells remains poorly understood. Recent work in this area, utilizing a peptide substrate as a probe for cellular TACE activity, indicates that the intrinsic activity of the enzyme is somehow increased.


Natriuretic peptide receptor 3 (NPR3) is regulated by microRNA-100.
Natriuretic peptide receptor 3 (NPR3) is the clearance receptor for the cardiac natriuretic peptides (NPs). By modulating the level of NPs, NPR3 plays an important role in cardiovascular homeostasis. Although the physiological functions of NPR3 have been explored, little is known about its regulation in health or disease. MicroRNAs play an essential role in the post-transcriptional expression of many genes. Our aim was to investigate potential microRNA-based regulation of NPR3 in multiple models. Hypoxic challenge elevated levels of NPPB and ADM mRNA, as well as NT-proBNP and MR-proADM in human left ventricle derived cardiac cells (HCMa), and in the corresponding conditioned medium, as revealed by qRT-PCR and ELISA. NPR3 was decreased while NPR1 was increased by hypoxia at mRNA and protein levels in HCMa. Down-regulation of NPR3 mRNA was also observed in infarct and peri-infarct cardiac tissue from rats undergoing myocardial infarction. From microRNA microarray analyses and microRNA target predictive databases, miR-100 was selected as a candidate regulator of NPR3 expression. Further analyses confirmed up-regulation of miR-100 in hypoxic cells and associated conditioned media. Antagomir-based silencing of miR-100 enhanced NPR3 expression in HCMa. Furthermore, miR-100 levels were markedly up-regulated in rat hearts and in peripheral blood after myocardial infarction and in the blood from heart failure patients. Results from this study point to a role for miR-100 in the regulation of NPR3 expression, and suggest a possible therapeutic target for modulation of NP bioactivity in heart disease.


The effect of climbing ability and slope inclination on vertical foot loading using a novel force sensor instrumentation system.
The aim of the study was to assess the effects of climbing ability and slope inclination on vertical loading both in terms the forces involved and physiological responses. Five novice and six intermediate female climbers completed a climbing route at three slope inclinations (85°, 90°, and 98°). The vertical loading during the climb was assessed by force-time integral using a Novel Pedar-X insole and physiological responses via oxygen uptake and heart rate. The novice climbers had a significantly lower (p < 0.05) vertical loading on foot holds and higher oxygen uptake and heart rate compared to intermediate climbers. A significant negative correlation was identified between the force-time integral and oxygen uptake (R = -0.72), and with heart rate (R = -0.64), respectively. The time-force integral decreased across the ascents with increasing slope inclination (p < 0.001). The results indicate that more advanced ability climbers make greater use of foot holds, with associated lowering in physiological response (oxygen uptake and heart rate) across all slope inclinations.


Isoform diversity, regulation, and functional adaptation of troponin and calponin.
Actin-activated myosin II motor function powers muscle contraction and nonmuscle cell motility. The actin-myosin-derived contractility has evolved with a great diversity in different muscle and cell types. Actin filament-based regulation controls striated muscle contraction and plays a role in modulating smooth muscle contractility and nonmuscle cell motility. This review focuses on the isoform diversity and functional adaptations of troponin in striated muscle and calponin in smooth muscle and nonmuscle cells. The gene regulation, alternative RNA splicing, and posttranslational modifications of troponin I and troponin T are summarized, together with recent progress in calponin studies. The biologic significance of the structural and functional diversity and regulation of troponin and calponin is discussed for roles in normal contractility and diseases.


Physical Frailty, Cognitive Impairment, and the Risk of Neurocognitive Disorder in the Singapore Longitudinal Ageing Studies.
The independent and combined effects of physical and cognitive domains of frailty in predicting the development of mild cognitive impairment (MCI) or dementia are not firmly established. This study included cross-sectional and longitudinal analyses of physical frailty (Cardiovascular Health Study criteria), cognitive impairment (Mini-Mental State Examination [MMSE]), and neurocognitive disorder (DSM-5 criteria) among 1,575 community-living Chinese older adults from the Singapore Longitudinal Ageing Studies. At baseline, 2% were frail, 32% were prefrail, and 9% had cognitive impairment (MMSE score < 23). Frailty at baseline was significantly associated with prevalent cognitive impairment. Physical frailty categories were not significantly associated with incident NCD, but continuous physical frailty score and MMSE score showed significant individual and joint associations with incident mild NCD and dementia. Compared with those who were robust and cognitively normal, prefrail or frail old adults without cognitive impairment had no increased risk of incident NCD, but elevated odds of association with incident NCD were observed for robust with cognitive impairment (odds ratio [OR] = 4.04, p < .001), prefrail with cognitive impairment (OR = 2.22, p = .044), and especially for frail with cognitive impairment (OR = 6.37, p = .005). The prevalence of co-existing frailty and cognitive impairment (cognitive frailty) was 1% (95% confidence interval [CI]: 0.5-1.4), but was higher among participants aged 75 and older at 5.0% (95% CI: 1.8-8.1). Physical frailty is associated with increased prevalence and incidence of cognitive impairment, and co-existing physical frailty and cognitive impairment confers additionally greater risk of incident NCD.


Atomistic Model for Nearly Quantitative Simulations of Langmuir Monolayers.
Lung surfactant and a tear film lipid layer are examples of biologically relevant macromolecular structures found at the air-water interface. Because of their complexity, they are often studied in terms of simplified lipid layers, the simplest example being a Langmuir monolayer. Given the profound biological significance of these lipid assemblies, there is a need to understand their structure and dynamics on the nanoscale, yet there are not many techniques able to provide this information. Atomistic molecular dynamics simulations would be a tool fit for this purpose; however, the simulation models suggested until now have been qualitative instead of quantitative. This limitation has mainly stemmed from the challenge to correctly describe the surface tension of water with simulation parameters compatible with other biomolecules. In this work, we show that this limitation can be overcome by using the recently introduced four-point OPC water model, whose surface tension for water is demonstrated to be quantitatively consistent with experimental data and which is also shown to be compatible with the commonly employed lipid models. We further establish that the approach of combining the OPC four-point water model with the CHARMM36 lipid force field provides nearly quantitative agreement with experiments for the surface pressure-area isotherm for POPC and DPPC monolayers, also including the experimentally observed phase coexistence in a DPPC monolayer. The simulation models reported in this work pave the way for nearly quantitative atomistic studies of lipid-rich biological structures at air-water interfaces.


[Developmental bilharziasis caused by Schistosoma mansoni discovered 37 years after infestation].
The authors report a case of Mansoni Schistosomiasis whose course in always running thirty-seven years after the first and alone infestation in a fifty-six old man who come back from Madagascar in February 1948 and lives in France since this period eggs of schistosomiasis were living in a biopsy of rectal and sigmoid mucosea. Problem of adult worms longevity and clinical importance of such prolonged inapparent schistosomiasis are discussed.


Influence of an antioxidant vitamin-enriched drink on pre- and post-exercise lymphocyte antioxidant system.
Our aim was to determine whether the consumption of a functional drink enriched with vitamin C and vitamin E could alleviate exercise-induced oxidative damage in lymphocytes. Fourteen male amateur trained runners (32-36 years old) were randomized to receive antioxidant supplementation (152 mg vitamin C/day and 50 mg vitamin E/day) or placebo for 1 month in a double-blind study, and participated in a half-marathon race. The volunteers did not take any other antioxidant supplements. Blood samples were taken before and after the half-marathon and after 3 h recovery. Exercise increased lymphocyte malondialdehyde levels only in the placebo group (p < 0.001). Lymphocytes from the supplemented group accumulated more vitamin C and vitamin E than placebo after exercise and recovery (p < 0.05). Catalase and glutathione peroxidase activities increased only in the supplemented group after exercise, whereas superoxide dismutase activity increased in both groups (p < 0.05). Heme oxygenase-1 expression was enhanced during the recovery period only in the placebo group (p < 0.01). Bcl-2 expression decreased after exercise only in the placebo group and remained low during the recovery (p < 0.001). Intense exercise increases lipid peroxidation, decreases Bcl-2 expression and induces an antioxidant response in lymphocytes. Supplementation with moderate levels of antioxidant vitamins reduces exercise-induced oxidative damage, but without blocking the cellular adaptation to exercise.


Single channel studies of the phosphorylation of K+ channels in the squid giant axon. I. Steady-state conditions.
Phosphorylation of the delayed rectifier channel of squid potentiates the macroscopic K+ current and slows its activation kinetics. We have studied this phenomenon at the single channel level using the cut-open axon technique under steady-state conditions. In 10 mM external K+/310 mM internal K+ there are predominantly two types of channels present, a 20-pS and a 40-pS channel. In steady state at depolarized potentials, the 40-pS channel was most active, whereas the 20-pS channel tended to disappear due to a slow inactivation process. Two methods were developed to shift the population of channels toward a dephosphorylated state. One method consisted of predialyzing a whole axon with solutions containing no ATP, while recording the currents under axial-wire voltage clamp. A piece of axon was then removed and cut open, and single channel currents were recorded from the cut-open axon. A second method was based on the difference in diffusion coefficients for ATP and proteins such as the endogenous phosphatase. The axon was cut open in a solution that did not contain Ca2+ or Cl- in order to maintain the axoplasm structurally intact and permit endogenous phosphatase to act on the membrane while ATP diffused away, before removing the axoplasm and forming a membrane patch. When dephosphorylating conditions were used, the steady-state open probability of the 40-pS channel at 42 mV was very low (less than 0.0002), and the channel openings appeared as a series of infrequent, short-duration events. The channel activity was increased up to 150-fold by photoreleasing caged ATP inside the patch pipette in the presence of the catalytic subunit of protein kinase A. The sharp increase in open probability could be accounted for by a decrease of the slow component of the closed time distribution from 23 s to 170 ms with little change in the distribution of open times (1-2 ms) and no change in the single channel current amplitude. In voltage-jump experiments the contribution of the 40-pS channel to the delayed rectifier current was often small due to the large values of the latency to the first opening.


Mechanical design of DNA nanostructures.
Structural DNA nanotechnology is a rapidly emerging field that has demonstrated great potential for applications such as single molecule sensing, drug delivery, and templating molecular components. As the applications of DNA nanotechnology expand, a consideration of their mechanical behavior is becoming essential to understand how these structures will respond to physical interactions. This review considers three major avenues of recent progress in this area: (1) measuring and designing mechanical properties of DNA nanostructures, (2) designing complex nanostructures based on imposed mechanical stresses, and (3) designing and controlling structurally dynamic nanostructures. This work has laid the foundation for mechanically active nanomachines that can generate, transmit, and respond to physical cues in molecular systems.


[Treatment of pediatric obstructive sleep apnea hypopnea syndrome].
To evaluate treatment options of pediatric obstructive sleep apnea hypopnea syndrome (OSAHS) and their curative effect. In this study, there are 59 confirmed pediatric obstructive sleep apnea hypopnea syndrome cases diagnosed by polysomnography (PSG). Among them, 54 cases were treated by tonsillectomy or/and adenoidectomy directed by transoral endoscopy. Continue positive airway pressure (CPAP) was used for 2 cases in long-term. Three cases were treated with other conservative therapy. OSA-18 (quality of life for children with obstructive sleep apnea 18 items) was adopted as a disease-specific quality-of-life follow-up survey before and after treatment. During perioperation period, no postoperative bleeding or acute respiratory obstruction occurred. Follow-up sleep study was carried out 12-18 months postoperatively, no complications of velopharyngeal insufficiency, nasopharyngeal stenosis or residual adenoid were found. OSA-18 scores showed that quality of life had been significant improved after operation in 76.3% (45 cases), sleep disturbance in 88.1% (52 cases) and physical symptoms in 67.8% (40 cases). The pressure level of long-term CPAP in 2 cases was about 5.6-7.8 cmH2O. 3 cases were slightly improved treated with conservative therapy. Surgical removal of upper airway obstruction caused by enlargement of tonsil and adenoid is one of the most effective treatment for pediatric OSAHS. Fibrous endoscopy and cephalometric analysis are helpful to confirm surgical indication. The advantages of transoral endoscopy directed adenoidectomy are as follows: clear and direct visualization, complete removal, without damaging of the peripheral structures.


An unusual nitrofurantoin-induced drug reaction.
A case of drug-induced lupus erythematosus associated with erythema multiforme due to nitrofurantoin is presented. This case differs from those previously reported in that no pulmonary infiltrate was found. In 21 months, there was a complete clinical and serologic remission after nitrofurantoin was withdrawn from the therapeutic regimen.


Operant control of epileptic neurons in chronic foci of monkeys.
(1) Six Macaca mulatta monkeys were rendered chronically epileptic by subpial alumina injection in left precentral cortex. The monkeys were then trained to operantly control the firing patterns of single units from the epileptic focus and contralateral homotopic cortex using a standardized paradigm. In 30 experiments, units were recorded simultaneously from both hemispheres. (2) For epileptic and normal neurons, there are no consistent relationships between cortical EEG spikes and unit firing. (3) Previous parameters for quantifying single unit epileptogenicity are not adequate for defining the complex spectrum and relative degree of unit abnormalities within the chronic focus. A primary difference between normal and epileptic neurons is that monkeys can modify the firing pattern and rate of the former but can modify, only slightly, the integrated firing rate of the latter. (4) A previously described 'non-burst' epileptiform firing pattern consisting of 8--18 msec doublets is investigated in further detail. This pattern may represent units along a continuum between normalcy and long-first-interval burst firing. (5) The majority of epileptic units' firing patterns become more normal when the monkey is reinforced to control the firing pattern of a contralateral unit. Then, if the monkey is reinforced to control the epileptic unit, subtle abnormal firing patterns become more apparent. Therefore, the firing patterns of epileptic units are effected in two ways: the non-specific effect of alerting to the operant task, and the more specific effect that accompanies the monkey's attempts to attain reinforcement when it is contingent on the behavior of the specific neuron. (6) Although epileptiform EEG spikes are projected to homotopic contralateral precentral cortex, there is no evidence to suggest that single units within such cortex fire in patterns correlative with intrinsic epileptogenic burst generating properties. Therefore, if the 'mirror focus' is defined as cortex which has become autonomously epileptogenic secondary to persistent transsynaptic activity from a primary focus, these data do not support the concept that mirror foci develop in primate precentral cortex.


Functional endoscopy techniques for tracking stem cell fate.
Tracking and monitoring implanted stem cells are essential to maximize benefits and to minimize the side effects of stem cell therapy for personalized or "precision" medicine. Previously, we proposed a comprehensive biological Global Positioning System (bGPS) to track and monitor stem cells in vivo. Magnetic resonance imaging (MRI), positron emission tomography (PET), bioluminescent imaging, fluorescence imaging, and single-photon emission computerized tomography (SPECT) have been utilized to track labeled or genetically-modified cells in vivo in rats and humans. A large amount of research has been dedicated to the design of reporter genes and molecular probes for imaging and the visualization of the biodistribution of the implanted cells in high resolution. On the other hand, optical-based functional imaging, such as photoacoustic imaging (PAI), optical coherence tomography (OCT), and multiphoton microscopy (MPM), has been implemented into small endoscopes to image cells inside the body. The optical fiber allows miniaturization of the imaging probe while maintaining high resolution due to light-based imaging. Upon summarizing the recent progress in the design and application of functional endoscopy techniques for stem cell monitoring, we offer perspectives for the future development of endoscopic molecular imaging tools for in vivo tracking of spatiotemporal changes in subclonal evolution at the single cell level.


Effects of chitosan and whole raw soybeans on ruminal fermentation and bacterial populations, and milk fatty acid profile in dairy cows.
The objective of this study was to evaluate whether providing chitosan (CHI) to cows fed diets supplemented with whole raw soybeans (WRS) would affect the nutrient intake and digestibility, ruminal fermentation and bacterial populations, microbial protein synthesis, N utilization, blood metabolites, and milk yield and composition of dairy cows. Twenty-four multiparous Holstein cows (141 ± 37.1 d in milk, 38.8 ± 6.42 kg/d of milk yield; mean ± SD) were enrolled to a 4 × 4 Latin square design experiment with 23-d periods. Cows were blocked within Latin squares according to milk yield, days in milk, body weight, and rumen cannula (n = 8). A 2 × 2 factorial treatment arrangement was randomly assigned to cows within blocks. Treatments were composed of diets with 2 inclusion rates of WRS (0 or 14% diet dry matter) and 2 doses of CHI (0 or 4 g/kg of dry matter, Polymar Ciência e Nutrição, Fortaleza, Brazil). In general, CHI+WRS negatively affected nutrient intake and digestibility of cows, decreasing milk yield and solids production. The CHI increased ruminal pH and decreased acetate to propionate ratio, and WRS reduced NH3-N concentration and acetate to propionate in the rumen. The CHI reduced the relative bacterial population of Butyrivibrio group, whereas WRS decreased the relative bacterial population of Butyrivibrio group, and Fibrobacter succinogenes, and increased the relative bacterial population of Streptococcus bovis. No interaction effects between CHI and WRS were observed on ruminal fermentation and bacterial populations. The CHI+WRS decreased N intake, microbial N synthesis, and N secreted in milk of cows. The WRS increased N excreted in feces and consequently decreased the N excreted in urine. The CHI had no effects on blood metabolites, but WRS decreased blood concentrations of glucose and increased blood cholesterol concentration. The CHI and WRS improved efficiency of milk yield of cows in terms of fat-corrected milk, energy-corrected milk, and net energy of lactation. The CHI increased milk concentration [g/100 g of fatty acids (FA)] of 18:1 trans-11, 18:2 cis-9,cis-12, 18:3 cis-9,cis-12,cis-15, 18:1 cis-9,trans-11, total monounsaturated FA, and total polyunsaturated FA. The WRS increased total monounsaturated FA, polyunsaturated FA, and 18:0 to unsaturated FA ratio in milk of cows. Evidence indicates that supplementing diets with unsaturated fat sources along with CHI negatively affects nutrient intake and digestibility of cows, resulting in less milk production. Diet supplementation with CHI or WRS can improve feed efficiency and increases unsaturated FA concentration in milk of dairy cows.


Precision nutrition: hype or hope for public health interventions to reduce obesity?
High-income countries are experiencing an obesity epidemic that follows a socioeconomic gradient, affecting groups of lower socioeconomic status disproportionately. Recent clinical findings have suggested new perspectives for the prevention and treatment of obesity, using personalized dietary approaches. Precision nutrition (PN), also called personalized nutrition, has been developed to deliver more preventive and practical dietary advice than 'one-size-fits-all' guidelines. With interventions becoming increasingly plausible at a large scale thanks to artificial intelligence and smartphone applications, some have begun to view PN as a novel way to deliver the right dietary intervention to the right population. We argue that large-scale PN, if taken alone, might be of limited interest from a public health perspective. Building on Geoffrey Rose's theory regarding the differences in individual and population causes of disease, we show that large-scale PN can only address some individual causes of obesity (causes of cases). This individual-centred approach is likely to have a small impact on the distribution of obesity at a population level because it ignores the population causes of obesity (causes of incidence). The latter are embedded in the populations' social, cultural, economic and political contexts that make environments obesogenic. Additionally, the most socially privileged groups in the population are the most likely to respond to large-scale PN interventions. This could have the undesirable effect of widening social inequalities in obesity. We caution public health actors that interventions based only on large-scale PN are unlikely, despite current expectations, to improve dietary intake or reduce obesity at a population level.


How many replicates to accurately estimate fish biodiversity using environmental DNA on coral reefs?
Quantifying fish species diversity in rich tropical marine environments remains challenging. Environmental DNA (eDNA) metabarcoding is a promising tool to face this challenge through the filtering, amplification, and sequencing of DNA traces from water samples. However, because eDNA concentration is low in marine environments, the reliability of eDNA to detect species diversity can be limited. Using an eDNA metabarcoding approach to identify fish Molecular Taxonomic Units (MOTUs) with a single 12S marker, we aimed to assess how the number of sampling replicates and filtered water volume affect biodiversity estimates. We used a paired sampling design of 30 L per replicate on 68 reef transects from 8 sites in 3 tropical regions. We quantified local and regional sampling variability by comparing MOTU richness, compositional turnover, and compositional nestedness. We found strong turnover of MOTUs between replicated pairs of samples undertaken in the same location, time, and conditions. Paired samples contained non-overlapping assemblages rather than subsets of one another. As a result, non-saturated localized diversity accumulation curves suggest that even 6 replicates (180 L) in the same location can underestimate local diversity (for an area <1 km). However, sampling regional diversity using ~25 replicates in variable locations (often covering 10 s of km) often saturated biodiversity accumulation curves. Our results demonstrate variability of diversity estimates possibly arising from heterogeneous distribution of eDNA in seawater, highly skewed frequencies of eDNA traces per MOTU, in addition to variability in eDNA processing. This high compositional variability has consequences for using eDNA to monitor temporal and spatial biodiversity changes in local assemblages. Avoiding false-negative detections in future biomonitoring efforts requires increasing replicates or sampled water volume to better inform management of marine biodiversity using eDNA.


Patterns of smokeless tobacco use in a population of high school students.
We assessed the prevalence of smokeless tobacco use by 901 high school students in two Arkansas communities, and identified factors associated with initiation and maintenance. Results showed that 36.7 per cent of males and 2.2 per cent of females currently used smokeless tobacco. Modal responses indicated that if smokeless tobacco was used at all, it was used with frequency on an almost daily basis. Health professionals need to monitor this phenomenon closely and take appropriate preventive actions.


Neuronal intranuclear inclusion disease and juvenile parkinsonism.
Juvenile parkinsonism (onset age <20 yrs) is uncommon and few cases with neuropathologic confirmation have been reported. We present the case of a 17-year-old boy who presented with asymmetric arm tremor and bulbar symptoms. His paternal great aunt had parkinsonism with onset at age 22 years. Examination revealed parkinsonism in the absence of additional neurologic signs except for delayed pupillary responses to light. He responded well to levodopa but developed motor fluctuations and disabling dyskinesias after 3 years of treatment. Following attempted withdrawal of levodopa at age 24 years, he developed severe aspiration pneumonia complicated by cardiorepiratory arrests and he died 6 months later. At autopsy, the dominant histologic feature was wide-spread neuronal hyaline intranuclear inclusions. Neuronal depletion was observed in the substantia nigra, locus ceruleus, and, to a lesser extent, in the frontal cortex, and inclusions were particularly prominent in these areas. Inclusions were immunoreactive for ubiquitin and were typical of those seen in neuronal intranuclear inclusion disease (NIID), a rare, multisytem neurodegenerative disease. NIID should be considered in the differential diagnosis of juvenile parkinsonism. A link between NIID and hereditary neurodegenerative disorders characterized by expanded polyglutamine tracts is supported by the similar appearance of intranuclear inclusions in both conditions and by a family history in some cases of NIID.


Studies on SP6 promoter using a new plasmid vector that allows gene insertion at the transcription initiation site.
All the phage-promoter containing subcloning vectors available for in vitro transcription reactions contain a polylinker away from the transcription initiation site. A new SP6 transcription subcloning vector, pCKSP6, has been constructed, in which a gene can be inserted precisely at the transcription initiation site. This was achieved by bringing the BamHI cleavage site into the initiation site. When DNA ends of both insert gene and BamHI cleaved pCKSP6 are made blunt-ended using a single strand specific nuclease, the in vitro transcripts of the recombinant DNA by SP6 RNA polymerase will contain only the gene sequence immediately after the initiation base G. Mung bean nuclease was used to generate a series of mutants resulting from step-wise deletion of single base pairs around the initiation site. Transcription assays with these SP6 promoter mutants revealed that not only the sequence immediately upstream of the initiation site but also the six base pairs from position +1 to +6 are important elements for promoter binding and/or transcription initiation activity. Furthermore, there appears to be a hierarchy of importance of each base pair in the order of position +1 greater than +2 greater than +3 greater than +4, +5, +6, -1, -2.


Combined Transplantation of Human Neuronal and Mesenchymal Stem Cells following Spinal Cord Injury.
Transplantation of human fetal neural stem cells (hNSCs) previously demonstrated significant functional recovery after spinal cord contusion in rats. Other studies indicated that human mesenchymal stem cells (hMSCs) can home to areas of damage and cross the blood-brain barrier. The purpose of this article is to determine if combined administration of mesenchymal stem cells and neuronal stem cells improves functional outcomes in rats. The study design was a randomized controlled animal trial. Female adult Long-Evans hooded rats underwent laminectomy at T10 level. Moderate spinal cord contusion at T10 level was induced by the MASCIS Impactor. Four groups were identified. The MSC + NSC group received hMSCs intravenously (IV) immediately after spinal cord injury (acute) and returned 1 week later (subacute) for injection of hNSC directly at site of injury. The MSC-only group received hMSC IV acutely and cell media subacutely. The NSC-only group received cell media IV acutely and hNSC subacutely. The control group received cell media IV acutely and subacutely. Subjects were assessed for 6 weeks using Basso, Beattie, Bresnahan Locomotor Rating Score. Twenty-four subjects were utilized, six subjects in each group. Statistically significant functional improvement was seen in the MSC + NSC group and the NSC-only group versus controls (p = 0.027, 0.042, respectively). The MSC-only group did not demonstrate a significant improvement over control (p = 0.145). Comparing the MSC + NSC group and the NSC-only group, there was no significant difference (p = 0.357). Subacute transplantation of hNSCs into contused spinal cord of rats led to significant functional recovery when injected either with or without acute IV administration of hMSCs. Neither hMSCs nor addition of hMSC to hNSC resulted in significant improvement.


Inflammation-induced up-regulation of ionotropic glutamate receptor expression in human skin.
N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methylisoxazolone-4-propionic acid (AMPA), and kainate (KA) receptors are members of the ionotropic glutamate receptor (iGluR) family and are increased in inflamed rat skin. These receptors contribute to inflammatory pain. In this study, we have examined whether there is a similar increase in iGluRs in inflamed human skin in the presence of inflammatory pain. Normal and inflamed-skin biopsies were obtained from eight patients undergoing elective wound-debridement surgery. Real-time-polymerase chain reaction (PCR) and western blot analysis were used for quantitation of iGluR mRNA and protein in normal and inflamed human skin. A significant increase in mRNA and protein for NMDA, AMPA, and KA receptor subunits was detected in inflamed compared with normal skin. The amounts of NMDA (NR1 subunit), AMPA (GluR2 subunit), and KA (GluR6 subunit) mRNA in inflamed skin were mean 6 (SD 1.6-fold), 2.5 (0.6-fold), and 3.8 (0.9-fold) (P<0.05), respectively, greater than that measured in normal skin. The ratio of NR1, GluR2, and GluR6 protein in inflamed compared with normal skin was 5.7 (1.2), 2.4 (0.5), and 3.6 (0.9) (P<0.05), respectively. These results, in human tissue, demonstrate that iGluR mRNA and protein expression are increased during persistent inflammation and that this increased activity may be involved in mediating clinical inflammatory pain in human skin.


A Bayesian approach to quantifying the effects of mass poultry vaccination upon the spatial and temporal dynamics of H5N1 in Northern Vietnam.
Outbreaks of H5N1 in poultry in Vietnam continue to threaten the livelihoods of those reliant on poultry production whilst simultaneously posing a severe public health risk given the high mortality associated with human infection. Authorities have invested significant resources in order to control these outbreaks. Of particular interest is the decision, following a second wave of outbreaks, to move from a "stamping out" approach to the implementation of a nationwide mass vaccination campaign. Outbreaks which occurred around this shift in policy provide a unique opportunity to evaluate the relative effectiveness of these approaches and to help other countries make informed judgements when developing control strategies. Here we use Bayesian Markov Chain Monte Carlo (MCMC) data augmentation techniques to derive the first quantitative estimates of the impact of the vaccination campaign on the spread of outbreaks of H5N1 in northern Vietnam. We find a substantial decrease in the transmissibility of infection between communes following vaccination. This was coupled with a significant increase in the time from infection to detection of the outbreak. Using a cladistic approach we estimated that, according to the posterior mean effect of pruning the reconstructed epidemic tree, two thirds of the outbreaks in 2007 could be attributed to this decrease in the rate of reporting. The net impact of these two effects was a less intense but longer-lasting wave and, whilst not sufficient to prevent the sustained spread of outbreaks, an overall reduction in the likelihood of the transmission of infection between communes. These findings highlight the need for more effectively targeted surveillance in order to help ensure that the effective coverage achieved by mass vaccination is converted into a reduction in the likelihood of outbreaks occurring which is sufficient to control the spread of H5N1 in Vietnam.


Evaluation of Functional Outcomes after Stapes Surgery in Patients with Clinical Otosclerosis in a Teaching Institution.
Introduction Otosclerosis is a primary disease of the temporal bone that leads to stapes ankylosis. Hearing loss is the main symptom. Treatment includes surgery, medical treatment, and sound amplification therapy alone or in combination. Objective To evaluate the functional outcomes of patients with clinical diagnosis of otosclerosis undergoing primary stapes surgery in a teaching institution. Method Retrospective descriptive study. Results A total of 210 ears of 163 patients underwent stapes surgery. Of the 163 patients, 116 (71.2%) underwent unilateral surgery and 47 (28.8%) underwent bilateral surgery. Six of the 210 operated ears had obliterative otosclerosis. The average preoperative and postoperative air-bone gap was 32.06 and 4.39 dB, respectively. The mean preoperative and postoperative bone conduction threshold was 23.17 and 19.82 dB, respectively. A total of 184 (87.6%) ears had a residual air-bone gap <10 dB, and 196 (93.3%) had a residual air-bone gap ≤15 dB. Two patients (0.95%) had severe sensorineural hearing loss. Conclusion Stapes surgery showed excellent functional hearing outcomes in this study. This surgery may be performed in educational institutions with the supervision of experienced surgeons.


Quantitative immunofluorescence microscopy of subcellular GLUT4 distribution in human skeletal muscle: effects of endurance and sprint interval training.
Increases in insulin-mediated glucose uptake following endurance training (ET) and sprint interval training (SIT) have in part been attributed to concomitant increases in glucose transporter 4 (GLUT4) protein content in skeletal muscle. This study used an immunofluorescence microscopy method to investigate changes in subcellular GLUT4 distribution and content following ET and SIT. Percutaneous muscle biopsy samples were taken from the m. vastus lateralis of 16 sedentary males in the overnight fasted state before and after 6 weeks of ET and SIT. An antibody was fully validated and used to show large (> 1 μm) and smaller (<1 μm) GLUT4-containing clusters. The large clusters likely represent trans-Golgi network stores and the smaller clusters endosomal stores and GLUT4 storage vesicles (GSVs). Density of GLUT4 clusters was higher at the fibre periphery especially in perinuclear regions. A less dense punctate distribution was seen in the rest of the muscle fibre. Total GLUT4 fluorescence intensity increased in type I and type II fibres following both ET and SIT. Large GLUT4 clusters increased in number and size in both type I and type II fibres, while the smaller clusters increased in size. The greatest increases in GLUT4 fluorescence intensity occurred within the 1 μm layer immediately adjacent to the PM. The increase in peripheral localisation and protein content of GLUT4 following ET and SIT is likely to contribute to the improvements in glucose homeostasis observed after both training modes.


Behavioral responses of big brown bats to dives by praying mantises.
Insectivorous echolocating bats face a formidable array of defenses employed by their airborne prey. One such insect defense is the ultrasound-triggered dive, which is a sudden, rapid drop in altitude, sometimes all the way to the ground. Although many previous studies have investigated the dynamics of such dives and their effect on insect survival rate, there has been little work on how bats may adapt to such an insect defense employed in the middle of pursuit. In this study we investigated how big brown bats (Eptesicus fuscus) adjust their pursuit strategy when flying praying mantises (Parasphendale agrionina) execute evasive, ultrasound-triggered dives. Although the mantis dive occasionally forced the bat to completely abort its chase (25% trials), in a number of cases (75% trials) the bat followed the mantis into the dive. In such cases the bat kept its sonar beam locked onto the target and maneuvered to maintain the same time efficient strategy it adopted during level flight pursuit, though it was ultimately defeated by the dive. This study suggests that although the mantis dive can be effective in evading the bat, it does not always deter the bat from continuing pursuit and, given enough altitude, the bat can potentially capture diving prey using the same flight strategy it employs to intercept prey in level flight.


No significant association between anesthesia group concentration and private insurer payments in the United States.
Markets for physician services are becoming increasingly concentrated, with many areas being dominated by a few groups. Antitrust authorities are concerned that increasing concentration will lead to inappropriately high payments for physician services from private insurers. The authors examined the association between market concentration and private insurer payments for anesthesia services. The authors obtained data on average payments from private insurers for five commonly used anesthesia Current Procedure Terminology codes for physicians located in 229 counties in the United States between 2002 and 2010. The authors calculated a measure of market concentration (the Herfindahl-Hirschman Index [HHI]) for anesthesiologists in each county using Medicare claims data. The authors then estimated the association between market concentration and private insurer payments using a difference-in-differences approach to minimize confounding. Private insurer payments to anesthesiologists in more concentrated markets were not significantly different from payments in less concentrated markets. Compared with the 25% of counties with the least concentration (counties with an HHI in the 0th to 25th percentile), payments in counties in the 25th to 50th percentile of HHI were approximately 0.51% less (95% CI, -2.3 to 1.3%, P = 0.95), whereas payments in counties in the 50th to 75th percentile of HHI were approximately 2.8% less (95% CI, -6.7 to 1.4%, P = 0.41) and payments in counties in the 75th to 100th percentile were approximately 3.1% less (95% CI, -8.1 to 1.2%, P = 0.32). Increasing market concentration of anesthesia groups is not associated with significantly greater payments from private insurers.


Sargachromenol Attenuates Inflammatory Responses by Regulating NF-κB and Nrf2 Pathways in RAW 264.7 Cells and LPS-treated Mice.
This study aims to explore the anti-inflammatory mechanisms of sargachromenol in both RAW 264.7 cells and lipopolysaccharide (LPS)-treated mice, as previous reports have suggested that sargachromenol possesses anti-aging, anti-inflammatory, antioxidant, and neuroprotective properties. Although the precise mechanism behind its anti-inflammatory activity remains unclear, pretreatment with sargachromenol effectively reduced the production of nitric oxide, prostaglandin E2, and interleukin (IL)-1β in LPS-stimulated RAW 264.7 cells by inhibiting cyclooxygenase-2. Moreover, sargachromenol inhibited the activation of nuclear factor-κB (NF-κB) by preventing the degradation of the inhibitor of κB-α (IκB-α) and inhibiting protein kinase B (Akt) phosphorylation in LPS-stimulated cells. We also found that sargachromenol induced the production of heme oxygenase-1 (HO-1) by activating the nuclear transcription factor erythroid-2-related factor 2 (Nrf2). In LPS-treated mice, oral administration of sargachromenol effectively reduced the levels of IL-1β, IL-6, and tumor necrosis factor-α (TNF-α) in the serum, suggesting its ability to suppress the production of inflammatory mediators by inhibiting the Akt/NF-κB pathway and upregulating the Nrf2/HO-1 pathway.