Elevation of cytosolic calcium precedes anoxic gene expression in maize suspension-cultured cells.
Based on pharmacological evidence, we previously proposed that intracellular Ca2+ mediates the perception of O2 deprivation in maize seedlings. Herein, using fluorescence imaging and photometry of Ca2+ in maize suspension-cultured cells, the proposal was further investigated. Two complementary approaches were taken: (1) real time analysis of anoxia-induced changes in cytosolic Ca2+ concentration ([Ca]i) and (2) experimental manipulation of [Ca]i and then assay of the resultant anoxia-specific responses. O2 depletion caused an immediate increase in [Ca2+]i, and this was reversible within a few seconds of reoxygenation. The [Ca]i elevation proceeded independent of extracellular Ca2+. The kinetics of the Ca2+ response showed that it occurred much earlier than any detectable changes in gene expression. Ruthenium red blocked the anoxic [Ca]i elevation and also the induction of adh1 (encoding alcohol dehydrogenase) and sh1 (encoding sucrose synthase) mRNA. Ca2+, when added along with ruthenium red, prevented the effects of the antagonist on the anoxic responses. Verapamil and bepridil failed to block the [Ca]i rise induced by anoxia and were equally ineffective on anoxic gene expression. Caffeine induced an elevation of [Ca]i as well as ADH activity under normoxia. The data provide direct evidence for [Ca]i elevation in maize cells as a result of anoxia-induced mobilization of Ca2+ from intracellular stores. Furthermore, any manipulation that modified the [Ca]i rise brought about a parallel change in the expression of two anoxia-inducible genes. Thus, these results corroborate our proposal that [Ca]i is a physiological transducer of anoxia signals in plants.


A Single Surgeon's Experience with Open, Laparoscopic, and Robotic Partial Nephrectomy.
Objective. To report the perioperative outcomes of patients treated with partial nephrectomy by a single surgeon using three surgical modalities-open, laparoscopic, and robotic. Methods. Between August 2006 and February 2012, 106 consecutive patients underwent open partial nephrectomy (OPN) (n = 23), laparoscopic partial nephrectomy (LPN) (n = 48), and robotic partial nephrectomy (RPN) (n = 35) by a single surgeon. Clinical variables, operative parameters, and renal functional outcomes were analyzed. Results. Preoperative patient characteristics were similar except for baseline glomerular filtration rate (GFR), which was highest in the RPN group (P = 0.004). Surgery time was longest in the RPN group (244 minutes) and shortest in the OPN group (163 minutes, P < 0.0001). Patients who had OPN had the highest incidence of 30-day complications (30%), while the RPN approach had the lowest (14%, P = 0.008). Conclusions. When performed by a single surgeon, robotic partial nephrectomy appears to be associated with fewer complications than both open and laparoscopic partial nephrectomy. Kidney function was not affected by surgical approach.


Effects of friction and nonlinearities on the separation of arterial waves into their forward and backward components.
In this paper we examine the importance of fluid friction and nonlinearities due to the area-pressure relationship and to the convective acceleration on the separation of arterial pressure and flow waves into their forward and backward components. Experiments were run in straight uniform nonlinearly elastic tubes. Different degrees of fluid friction and nonlinearities, covering the physiological range, have been tested. We predicted the forward and backward running pressure components using two wave separation methods: the classical linear method (Westerhof et al., Cardiovasc. Res, 6,648-656, 1972) and the first order correction (FOC) method (Pythoud et al., Trans ASME J. Biomech. Engng, in press) which takes nonlinearities and fluid friction into account. We found that the two methods yield somewhat different predictions. The differences tend to increase with the degree of fluid friction and nonlinearities and are typically of the order of 4-8%. We further compared the transmission ratio of forward and backward waves predicted by both methods. The transmission ratio was found to be overestimated by 10% by the classical linear method. The nonlinear method gave more accurate estimates, consistent with theory. We conclude that, for in vivo applications, the classical linear method should be the method of choice because it is simpler to use and the erros involved (4-8%) are comparable to measurement erros.


Evidence synthesis and guideline development in genomic medicine: current status and future prospects.
With the accelerated implementation of genomic medicine, health-care providers will depend heavily on professional guidelines and recommendations. Because genomics affects many diseases across the life span, no single professional group covers the entirety of this rapidly developing field. To pursue a discussion of the minimal elements needed to develop evidence-based guidelines in genomics, the Centers for Disease Control and Prevention and the National Cancer Institute jointly held a workshop to engage representatives from 35 organizations with interest in genomics (13 of which make recommendations). The workshop explored methods used in evidence synthesis and guideline development and initiated a dialogue to compare these methods and to assess whether they are consistent with the Institute of Medicine report "Clinical Practice Guidelines We Can Trust." The participating organizations that develop guidelines or recommendations all had policies to manage guideline development and group membership, and processes to address conflicts of interests. However, there was wide variation in the reliance on external reviews, regular updating of recommendations, and use of systematic reviews to assess the strength of scientific evidence. Ongoing efforts are required to establish criteria for guideline development in genomic medicine as proposed by the Institute of Medicine.


Spontaneous Pneumomediastinum in Non-Asthmatic Children with Exercise-Induced Bronchoconstriction.
Subcutaneous emphysema can result from rupture of the respiratory or gastrointestinal systems, commonly occurring after trauma or surgery, as well as from rupture of alveoli as pneumothorax or pneumomediastinum. Spontaneous pneumomediastinum with subcutaneous emphysema is rare in children without chest or neck trauma. Here, we report 2 cases of spontaneous pneumomediastinum with exercise-induced bronchoconstriction. The first case is an 11-year-old boy who presented with neck pain after vigorous exercise. Radiography showed pneumomediastinum. The second case is a 15-year-old boy who presented with pleuritic chest pain and respiratory failure requiring intubation. We extensively investigated the possible causes of pneumomediastinum. Both patients had no history of trauma or asthma, and were diagnosed with exercise-induced bronchoconstriction. They were discharged after conservative treatment, without complication. Early recognition and investigation of serious conditions should be promptly done in spontaneous pneumomediastinum patients. Conservative treatment, extensive investigations of predisposing factors, and treatment are important.


Palatability and the micro-structure of feeding in humans: the appetizer effect.
The effects of manipulated palatability on eating were assessed in 54 human volunteers eating pasta with a tomato sauce, with palatability adjusted by the addition of three levels of oregano. Meals were divided into 2 min eating episodes separated by brief pauses during which subjects rated aspects of appetite. Both intake and eating rate were greatest in the most palatable condition (0.27% oregano), whereas the addition of 0.54% reduced intake and eating rate. Hunger ratings increased during the initial stages of the meal with 0.27% oregano, but fell throughout the meal in the other conditions, whereas fullness ratings increased similarly in all three conditions. Rated food attractiveness was greatest with 0.27% oregano, and least with 0.54% oregano, but declined similarly through the meal with all three foods. Initial ratings of palatability were similar to those for food attractiveness, but changes in palatability across meals varied between subjects. Male subjects consistently ate more and faster than females, but similar effects of manipulated palatability on intake and subjective appetite were seen in both sexes. These data are consistent with the idea that palatability increases intake through a positive-feedback reward mechanism, and offers a novel method for measuring these effects.


Neurodevelopmental Delay Diagnosis Rates Are Increased in a Region with Aerial Pesticide Application.
A number of studies have implicated pesticides in childhood developmental delay (DD) and autism spectrum disorder (ASD). The influence of the route of pesticide exposure on neurodevelopmental delay is not well defined. To study this factor, we examined ASD/DD diagnoses rates in an area near our regional medical center that employs yearly aerial pyrethroid pesticide applications to combat mosquito-borne encephalitis. The aim of this study was to determine if areas with aerial pesticide exposure had higher rates of ASD/DD diagnoses. This regional study identified higher rates of ASD/DD diagnoses in an area with aerial pesticides application. Zip codes with aerial pyrethroid exposure were 37% more likely to have higher rates of ASD/DD (adjusted RR = 1.37, 95% CI = 1.06-1.78, p = 0.02). A Poisson regression model controlling for regional characteristics (poverty, pesticide use, population density, and distance to medical center), subject characteristics (race and sex), and local birth characteristics (prematurity, low birthweight, and birth rates) identified a significant relationship between aerial pesticide use and ASD/DD rates. The relationship between pesticide application and human neurodevelopment deserves additional study to develop safe and effective methods of mosquito prevention, particularly as communities develop plans for Zika virus control.


Do lumbar magnetic resonance imaging changes predict neuropathic pain in patients with chronic non-specific low back pain?
The aim of this observational, cross-sectional study was to analyse lumbar magnetic resonance imaging (MRI) findings in patients with non-specific chronic low back pain (CLBP), and to evaluate any correlation with pain intensity and their capacity to predict neuropathic pain (NP) in these patients.Fifty-two patients with non-specific CLBP, between 21 and 62 years of age, 50% men, were investigated. Lumbar MRI was employed to assess disc degeneration, endplate changes, Modic changes, disc displacement, facet degeneration, foraminal stenosis and central lumbar spinal stenosis. The characteristics of pain were evaluated and patients were divided into 2 subgroups: with NP (24 patients) and without NP (28 patients), based on the results of a DN4-interview. Correlations between particular MRI changes and their relations to the intensity of pain were evaluated. Logistic regression was used to disclose predictors of NP.Lumbar spine degenerative features were frequent in patients with non-specific CLBP, with L4/5 the most affected level. A significant correlation emerged between the severity of degenerative changes in particular lumbar spine structures (correlation coefficient ranging between 0.325 and 0.573), while no correlation was found between severity of degenerative changes and pain. Multivariate logistic regression revealed only 2 independent predictors of NP - female sex (odds ratio [OR] = 11.9) and a mean pain intensity of ≥4.5 in the previous 4 weeks (OR = 13.1).Degenerative changes in the lumbar spine are frequent MRI findings, but do not correlate with the intensity of pain and do not predict NP. However, female sex and pain intensity do predict NP.


Magnetohydrodynamics flow of a nanofluid driven by a stretching/shrinking sheet with suction.
The present paper investigates the effect of a mathematical model describing the aforementioned process in which the ambient nanofluid in the presence of suction/injection and magnetic field are taken into consideration. The flow is induced by an infinite elastic sheet which is stretched along its own plane. The stretching/shrinking of the sheet is assumed to be proportional to the distance from the slit. The governing equations are reduced to a nonlinear ordinary differential equation by means of similarity transformation. The consequential nonlinear equation is solved analytically. Consequences show that the flow field can be divided into a near-field region and a far-field region. Suction on the surface plays an important role in the flow development in the near-field whereas the far-field is responsible mainly by stretching. The electromagnetic effect plays exactly the same role as the MHD, which is to reduce the horizontal flow resulting from stretching. It is shown that the behavior of the fluid flow changes with the change of the nanoparticles type. The present study throws light on the analytical solution of a class of laminar boundary layer equations arising in the stretching/shrinking sheet problem.


IMAGING DIAGNOSIS-MAGNETIC RESONANCE IMAGING FINDINGS IN A CAT WITH SYSTEMIC REACTIVE ANGIOENDOTHELIOMATOSIS.
A 10-year-old, castrated male domestic short-haired cat was presented with an acute history of seizures, lethargy, anorexia, vomiting, and dyspnea. Magnetic resonance imaging of the brain showed multifocal areas of gray matter T2-weighted hyperintensity. The lesions did not enhance with intravenous contrast. The cat was diagnosed at necropsy with feline systemic reactive angioendotheliomatosis, a rare vascular proliferative disorder for which a treatment has not yet been identified. This report is the first to describe associated magnetic resonance imaging changes for this disease.


[Case of consciousness disturbance following a fever with spreading lesion over the bilateral splenia of the corpus callosum on MRI].
We report a 66-year-old man with spreading lesion over the bilateral splenia of the corpus callosum shown on MRI. On admission, unknown fever and myoclonus-like involuntary movement in the left forefinger and middle finger were observed. There were no remarkably abnormal data in the serum, the cerebrospinal fluid and electroencephalogram. However, T2-weighted MRI revealed the high signal spreading over the bilateral splenia of the corpus callosum, while enhanced effects were not observed by Gd contrast on T1-weighted MRI. Diffusion and FLAIR MRI also showed the high signals limited to the same part of the splenia bilaterally as on T2-weighted MRI, discriminating it from other lacunar lesions and old cerebral infarctions. Neurological features, which were considered to be directly associated with the fresh lesion, were impairment of verbal and visual memories. Near the time the therapies including high-dose steroid were given, the consciousness of the patient worsened rapidly on a day-to-day basis and disseminated intravascular coagulation syndrome (DIC) also occurred. By administering low-molecule heparin, consciousness disturbance and involuntary movements recovered completely. Following the improved symptoms, FLAIR MRI showed a reduced level of the signal seven months after the onset. A demyelinating disease over the bilateral splenia of the corpus callosum should be considered as the final diagnosis. Follow-up of the changes in this case is expected to provide a more accurate diagnosis.


Critical Care Management of Patients with COVID-19: Early Experience in Thailand.
Since late December 2019, the world has been challenged with an outbreak of COVID-19. In Thailand, an upper middle-income country with a limited healthcare infrastructure and restricted human resources, nearly 3,000 confirmed COVID-19 cases have been reported as of early May 2020. Public health policies aimed at preventing new COVID-19 cases were very effective in halting the pandemic in Thailand. Case fatality in Thailand has been low (1.7%), at least in part due to early stratification according to risk of disease severity and timely initiation of supportive care with affordable measures. We present our initial experience with COVID-19 in Thailand, focusing on several aspects that may have played a crucial role in curtailment of the pandemic, and elements of care for severely ill COVID-19 patients, including stratification, isolation, and affordable diagnostic approaches and supportive care measures. We also discuss local considerations concerning some proposed experimental treatments.


Body mass index (BMI) does not predict responses to psilocybin.
Psilocybin is a serotonin type 2A (5-HT2A) receptor agonist and naturally occurring psychedelic. 5-HT2A receptor density is known to be associated with body mass index (BMI), however, the impact of this on psilocybin therapy has not been explored. While body weight-adjusted dosing is widely used, this imposes a practical and financial strain on the scalability of psychedelic therapy. This gap between evidence and practice is caused by the absence of studies clarifying the relationship between BMI, the acute psychedelic experience and long-term psychological outcomes. Data were pooled across three studies using a fixed 25 mg dose of psilocybin delivered in a therapeutic context to assess whether BMI predicts characteristics of the acute experience and changes in well-being 2 weeks later. Supplementing frequentist analysis with Bayes Factors has enabled for conclusions to be drawn regarding the null hypothesis. Results support the null hypothesis that BMI does not predict overall intensity of the altered state, mystical experiences, perceptual changes or emotional breakthroughs during the acute experience. There was weak evidence for greater 'dread of ego dissolution' in participants with lower BMI, however, further analysis suggested BMI did not meaningfully add to the combination of the other covariates (age, sex and study). While mystical-type experiences and emotional breakthroughs were strong predictors of improvements in well-being, BMI was not. These findings have important implications for our understanding of pharmacological and extra-pharmacological contributors to psychedelic-assisted therapy and for the standardization of a fixed therapeutic dose in psychedelic-assisted therapy.


Dreaming furiously? A sleep laboratory study on the dream content of people with Parkinson's disease and with or without rapid eye movement sleep behavior disorder.
Rapid eye movement (REM) sleep behavior disorder (RBD) has been related to altered, action-filled, vivid, and aggressive dream content, but research comparing the possible differences in dreams of Parkinson's disease (PD) patients with and without RBD is scarce. The dream content of PD patients with and without RBD was analyzed with specific focus on action-filledness, vividness, emotional valence, and threats. A total of 69 REM and NREM dream reports were collected in the sleep laboratory, 37 from nine PD patients with RBD and 32 from six PD patients without RBD. A content analysis of (1) action-filledness (actions and environmental events); (2) vividness (emotions and cognitive activity); (3) intensity of actions, events and emotions; (4) emotional valence, and (5) threatening events was performed on the transcripts. Altogether 563 dream elements expressing action-filledness and vividness were found. There were no significant between-group differences in the number or distribution of elements reflecting action-filledness or vividness, emotional valence or threats. In within-group analyses, PD patients with RBD had significantly more negative compared to positive dreams (p = 0.012) and compared to PD patients without RBD, a tendency to have more intense actions in their dreams (p = 0.066). Based on the results of this study, there are no major between-group differences in the action-filledness, vividness, or threat content of dreams of PD patients with and without RBD. However, within-group analyses revealed that dreams were more often negatively than positively toned in PD patients with RBD.


[A case of Ehlers-Danlos syndrome].
Ehlers-Danlos syndrome, an inherited connective tissue disorder, is characterized by skin hyperextensibility, joint hypermobility and skin fragility. The disease is at least classified into 10 types, I to X, based on clinical features, biochemical abnormalities of the connective tissue and the mode of inheritance. A case is reported of a 6-year, 8-month-old girl who showed the features of the Ehlers-Danlos syndrome type VII. The findings were as follows: 1) Soft and velvety touch skin, characteristic joint hypermobility, talipes varus, and ventricular septal defects were present. 2) Widely set eyes, slight stabismus, epicanthal folds of the eyes, antimonogoloid slanted eyes, a broad bridge to the nose and low set ears were observed. 3) Gingival bleeding following toothbrushing, Enamel hypoplasia, high arched palate and laxity of the temporomandibular joint were present. 4) In the findings of the maxillary and mandibular dental arches, there was no tendency towards conical teeth in the deciduous dentition, and the width and length of the dental arches were remarkably narrow and short. 5) The roentogenographic findings revealed that there were no minor root deformities, pulpstones and calcification of the pulp. 6) The measurement of the width, length, and depth of the palate through moiré topography revealed that the palate was narrow and deep compared with the control. 7) As a result of cephalometric diagnosis, the nasion appeared to be protruded, therefore maxillary and mandibular seemed to be relatively retracted, and the gonial angle was obtuse. The dental management of patients with Ehlers-Danlos syndrome was discussed and reviewed.


Heparan sulfate mimetic PG545-mediated antilymphoma effects require TLR9-dependent NK cell activation.
Heparan sulfate (HS) is an essential component of the extracellular matrix (ECM), which serves as a barrier to tumor invasion and metastasis. Heparanase promotes tumor growth by cleaving HS chains of proteoglycan and releasing HS-bound angiogenic growth factors and facilitates tumor invasion and metastasis by degrading the ECM. HS mimetics, such as PG545, have been developed as antitumor agents and are designed to suppress angiogenesis and metastasis by inhibiting heparanase and competing for the HS-binding domain of angiogenic growth factors. However, how PG545 exerts its antitumor effect remains incompletely defined. Here, using murine models of lymphoma, we determined that the antitumor effects of PG545 are critically dependent on NK cell activation and that NK cell activation by PG545 requires TLR9. We demonstrate that PG545 does not activate TLR9 directly but instead enhances TLR9 activation through the elevation of the TLR9 ligand CpG in DCs. Specifically, PG545 treatment resulted in CpG accumulation in the lysosomal compartment of DCs, leading to enhanced production of IL-12, which is essential for PG545-mediated NK cell activation. Overall, these results reveal that PG545 activates NK cells and that this activation is critical for the antitumor effect of PG545. Moreover, our findings may have important implications for improving NK cell-based antitumor therapies.


Intracellular trafficking of Brucella abortus in J774 macrophages.
Brucella abortus is a facultative intracellular bacterium capable of surviving inside professional and nonprofessional phagocytes. The microorganism remains in membrane-bound compartments that in several cell types resemble modified endoplasmic reticulum structures. To monitor the intracellular transport of B. abortus in macrophages, the kinetics of fusion of phagosomes with preformed lysosomes labeled with colloidal gold particles was observed by electron microscopy. The results indicated that phagosomes containing live B. abortus were reluctant to fuse with lysosomes. Furthermore, newly endocytosed material was not incorporated into these phagosomes. These observations indicate that the bacteria strongly affect the normal maturation process of macrophage phagosomes. However, after overnight incubation, a significant percentage of the microorganisms were found in large phagosomes containing gold particles, resembling phagolysosomes. Most of the Brucella bacteria present in phagolysosomes were not morphologically altered, suggesting that they can also resist the harsh conditions prevalent in this compartment. About 50% colocalization of B. abortus with LysoSensor, a weak base that accumulates in acidic compartments, was observed, indicating that the B. abortus bacteria do not prevent phagosome acidification. In contrast to what has been described for HeLa cells, only a minor percentage of the microorganisms were found in compartments labeled with monodansylcadaverine, a marker for autophagosomes, and with DiOC6 (3,3'-dihexyloxacarbocyanine iodide), a marker for the endoplasmic reticulum. These results indicate that B. abortus bacteria alter phagosome maturation in macrophages. However, acidification does occur in these phagosomes, and some of them can eventually mature to phagolysosomes.


Studies on a modulatory role for adenosine in antigen and arachidonic acid induced contractions of guinea pig airways.
The modulatory effects of adenosine and selected derivatives were examined on antigen and arachidonic acid (AA) induced contractions of indomethacin-treated tracheal spirals and lung parenchymal strips from actively sensitized guinea pigs. Adenosine (up to 2 X 10(-4) M) had no effect on antigen-induced contractions, but inhibited AA-induced contractions by 30-40% if added 30 min prior to challenge. The weak effect of adenosine suggests that endogenous adenosine may only have a limited modulatory role in allergic bronchospasm. 2-Chloroadenosine (10(-6)-10(-4) M) dose-dependently inhibited antigen- and AA-induced contractions of trachea, but was considerably less effective on parenchyma. The substituted adenosine derivatives, R-phenylisopropyladenosine (R-PIA) and 5'-(N-ethylcarboxamido)-adenosine (NECA), and the adenosine transport inhibitor, 6-[p-nitrobenzyl)thio]-9-beta-D-ribofuranosyl purine, were also active as modulators, but their activity was relatively weak and varied with the stimulus and the tissue. An order of potency for R-PIA, NECA, and 2-chloroadenosine could not be determined and 8-phenyltheophylline (10(-5) M) was not an effective inhibitor of the effects of adenosine or the adenosine derivatives. This suggests that adenosine and its derivatives may modulate cells through mechanisms other than activation of conventional A1 and A2 receptors. A lack of specificity for the adenosine derivatives must also be considered.


Effects of Victimization and Violence on Suicidal Ideation and Behaviors Among Sexual Minority and Heterosexual Adolescents.
Sexual minority youth (SMY) are at higher risk for victimization and suicide than are heterosexual youth (HY). Relatively little research has examined which types of victimization are most closely linked to suicide, which is necessary to develop targeted prevention interventions. The present study was conducted to address this deficit. The data come from the 2011 Chicago Youth Risk Behavior Survey (n = 1,907). Structural equation modeling (SEM) in Mplus evaluated the direct, indirect, and total effects of sexual orientation on a latent indicator of suicidal ideation and behaviors via seven types of victimization. Four indicators of victimization were school-specific (e.g., harassment due to sexual orientation or gender identity (SO/GID), bullying, threatened or injured with a weapon, and skipping school due to safety concerns), and three indicators assessed other types of victimization (e.g., electronic bullying, intimate partner violence, and sexual abuse). Thirteen percent of youth were classified as SMY. Significantly more SMY than HY reported suicidal ideation (27.95% vs. 13.64%), a suicide plan (22.78% vs. 12.36%), and at least one suicide attempt (29.92% vs. 12.43%) in the past year (all P < .001). A greater percentage of SMY reported SO/GID-related harassment, skipping school, electronic bullying, and sexual abuse. Sexual orientation was not directly related to suicidal ideation and behaviors in SEM. Rather, SMY's elevated risk of suicidality functioned indirectly through two forms of school-based victimization: being threatened or injured with a weapon (B = .19, SE = .09, P ≤ .05) and experiencing SO/GID-specific harassment (B = .40, SE = .15, P ≤ .01). There also was a trend for SMY to skip school as a strategy to reduce suicide risk. Although SMY experience higher rates of victimization than do HY, school-based victimization that involves weapons or is due to one's SO/GID appear to be the most deleterious. That SMY may skip school to reduce their risk of suicidal ideation and behaviors is problematic, and schools should be encouraged to enact and enforce policies that explicitly protect SMY from victimization.


A conserved proline residue in Dothideomycete Avr4 effector proteins is required to trigger a Cf-4-dependent hypersensitive response.
CfAvr4, a chitin-binding effector protein produced by the Dothideomycete tomato pathogen Cladosporium fulvum, protects the cell wall of this fungus against hydrolysis by secreted host chitinases during infection. However, in the presence of the Cf-4 immune receptor of tomato, CfAvr4 triggers a hypersensitive response (HR), which renders the pathogen avirulent. Recently, several orthologues of CfAvr4 have been identified from phylogenetically closely related species of Dothideomycete fungi. Of these, DsAvr4 from Dothistroma septosporum also triggers a Cf-4-dependent HR, but CaAvr4 and CbAvr4 from Cercospora apii and Cercospora beticola, respectively, do not. All, however, bind chitin. To identify the region(s) and specific amino acid residue(s) of CfAvr4 and DsAvr4 required to trigger a Cf-4-dependent HR, chimeric and mutant proteins, in which specific protein regions or single amino acid residues, respectively, were exchanged between CfAvr4 and CaAvr4 or DsAvr4 and CbAvr4, were tested for their ability to trigger an HR in Nicotiana benthamiana plants transgenic for the Cf-4 immune receptor gene. Based on this approach, a single region common to CfAvr4 and DsAvr4 was determined to carry a conserved proline residue necessary for the elicitation of this HR. In support of this result, a Cf-4-dependent HR was triggered by mutant CaAvr4 and CbAvr4 proteins carrying an arginine-to-proline substitution at this position. This study provides the first step in deciphering how Avr4 orthologues from different Dothideomycete fungi trigger a Cf-4-dependent HR.


Recurrent polyploid origins and chloroplast phylogeography in the Arabis holboellii complex (Brassicaceae).
Arabis holboellii is a North American member of the Brassicaceae that can reproduce via sex or apomixis. Previous studies have shown sexual individuals to be diploid, whilst apomictic individuals can be diploid (and aneuploid) or polyploid (typically 3x). Apomictic individuals can furthermore be facultative (i.e. both sexual and apomictic seed production in a single individual). Using flow cytometry, ploidy variation in 245 accessions of A. holboellii and A. drummondii from western North America and Greenland has been examined. Additionally, the chloroplast trnL intron region from each accession was sequenced for phylogenetic analysis of ploidy variation. Based upon 17 informative single nucleotide and insertion-deletion polymorphisms, we identified seven and 14 chloroplast haplotypes for A. drummondii and A. holboellii, respectively. Six of the haplotypes were found in both species. Ten of the chloroplast haplotypes were characterized by diploid, aneuploid, and triploid individuals, and thus we conclude that polyploidy has repeatedly and independently arisen within the species complex. As triploid individuals, which undergo normal meiosis, can only reproduce through apomixis, this may imply that the phenotype apomixis has also arisen multiple times. Arabis holboellii thus appears to have some predisposition to evolve apomictic reproduction.


Modelflow underestimates cardiac output in heat-stressed individuals.
An estimation of cardiac output can be obtained from arterial pressure waveforms using the Modelflow method. However, whether the assumptions associated with Modelflow calculations are accurate during whole body heating is unknown. This project tested the hypothesis that cardiac output obtained via Modelflow accurately tracks thermodilution-derived cardiac outputs during whole body heat stress. Acute changes of cardiac output were accomplished via lower-body negative pressure (LBNP) during normothermic and heat-stressed conditions. In nine healthy normotensive subjects, arterial pressure was measured via brachial artery cannulation and the volume-clamp method of the Finometer. Cardiac output was estimated from both pressure waveforms using the Modeflow method. In normothermic conditions, cardiac outputs estimated via Modelflow (arterial cannulation: 6.1 ± 1.0 l/min; Finometer 6.3 ± 1.3 l/min) were similar with cardiac outputs measured by thermodilution (6.4 ± 0.8 l/min). The subsequent reduction in cardiac output during LBNP was also similar among these methods. Whole body heat stress elevated internal temperature from 36.6 ± 0.3 to 37.8 ± 0.4°C and increased cardiac output from 6.4 ± 0.8 to 10.9 ± 2.0 l/min when evaluated with thermodilution (P < 0.001). However, the increase in cardiac output estimated from the Modelflow method for both arterial cannulation (2.3 ± 1.1 l/min) and Finometer (1.5 ± 1.2 l/min) was attenuated compared with thermodilution (4.5 ± 1.4 l/min, both P < 0.01). Finally, the reduction in cardiac output during LBNP while heat stressed was significantly attenuated for both Modelflow methods (cannulation: -1.8 ± 1.2 l/min, Finometer: -1.5 ± 0.9 l/min) compared with thermodilution (-3.8 ± 1.19 l/min). These results demonstrate that the Modelflow method, regardless of Finometer or direct arterial waveforms, underestimates cardiac output during heat stress and during subsequent reductions in cardiac output via LBNP.


Epidemiology, Patterns of Care, and Mortality for Patients With Acute Respiratory Distress Syndrome in Intensive Care Units in 50 Countries.
Limited information exists about the epidemiology, recognition, management, and outcomes of patients with the acute respiratory distress syndrome (ARDS). To evaluate intensive care unit (ICU) incidence and outcome of ARDS and to assess clinician recognition, ventilation management, and use of adjuncts-for example prone positioning-in routine clinical practice for patients fulfilling the ARDS Berlin Definition. The Large Observational Study to Understand the Global Impact of Severe Acute Respiratory Failure (LUNG SAFE) was an international, multicenter, prospective cohort study of patients undergoing invasive or noninvasive ventilation, conducted during 4 consecutive weeks in the winter of 2014 in a convenience sample of 459 ICUs from 50 countries across 5 continents. Acute respiratory distress syndrome. The primary outcome was ICU incidence of ARDS. Secondary outcomes included assessment of clinician recognition of ARDS, the application of ventilatory management, the use of adjunctive interventions in routine clinical practice, and clinical outcomes from ARDS. Of 29,144 patients admitted to participating ICUs, 3022 (10.4%) fulfilled ARDS criteria. Of these, 2377 patients developed ARDS in the first 48 hours and whose respiratory failure was managed with invasive mechanical ventilation. The period prevalence of mild ARDS was 30.0% (95% CI, 28.2%-31.9%); of moderate ARDS, 46.6% (95% CI, 44.5%-48.6%); and of severe ARDS, 23.4% (95% CI, 21.7%-25.2%). ARDS represented 0.42 cases per ICU bed over 4 weeks and represented 10.4% (95% CI, 10.0%-10.7%) of ICU admissions and 23.4% of patients requiring mechanical ventilation. Clinical recognition of ARDS ranged from 51.3% (95% CI, 47.5%-55.0%) in mild to 78.5% (95% CI, 74.8%-81.8%) in severe ARDS. Less than two-thirds of patients with ARDS received a tidal volume 8 of mL/kg or less of predicted body weight. Plateau pressure was measured in 40.1% (95% CI, 38.2-42.1), whereas 82.6% (95% CI, 81.0%-84.1%) received a positive end-expository pressure (PEEP) of less than 12 cm H2O. Prone positioning was used in 16.3% (95% CI, 13.7%-19.2%) of patients with severe ARDS. Clinician recognition of ARDS was associated with higher PEEP, greater use of neuromuscular blockade, and prone positioning. Hospital mortality was 34.9% (95% CI, 31.4%-38.5%) for those with mild, 40.3% (95% CI, 37.4%-43.3%) for those with moderate, and 46.1% (95% CI, 41.9%-50.4%) for those with severe ARDS. Among ICUs in 50 countries, the period prevalence of ARDS was 10.4% of ICU admissions. This syndrome appeared to be underrecognized and undertreated and associated with a high mortality rate. These findings indicate the potential for improvement in the management of patients with ARDS. clinicaltrials.gov Identifier: NCT02010073.


A unified model of atrioventricular nodal conduction predicts dynamic changes in Wenckebach periodicity.
The atrioventricular (AV) node responds in a complex fashion to changes in activation rate. A variety of approaches have been used to explain these dynamic AV nodal responses, but none has been able to account fully for AV nodal behavior. Three specific rate-dependent properties of the AV node have been described: 1) time-dependent recovery after excitation, 2) an effect of short cycles to advance recovery ("facilitation"), and 3) a gradual slowing of conduction in response to sustained, high-frequency activation ("fatigue"). We hypothesized that a model incorporating quantitative descriptors of all three processes might be able to account for a wide variety of AV nodal behaviors. Quantitative descriptors of AV nodal recovery, facilitation, and fatigue were developed based on AV nodal conduction changes during selective pacing protocols in seven autonomically blocked dogs. These descriptors were incorporated into a set of mathematical equations that define AV nodal conduction of any beat based on activation history. The equations were then applied to predict pacing-induced Wenckebach periodicity in each dog. Experimental data were obtained after nine to 19 step decreases in atrial cycle length into the Wenckebach zone in each animal. Observed behaviors included complex patterns of block, a progressive increase in the level of block over 5 minutes of rapid pacing, and periods of alternating patterns of block. The model accurately predicted the onset of AV block at each cycle length, the relation between conduction ratio and cycle length as a function of time, and the changing patterns of Wenckebach periodicity during sustained atrial pacing. All three terms of the model equation (describing recovery, facilitation, and fatigue) were essential to account fully for the observed behaviors. Elimination of AV nodal fatigue from the model resulted in failure to account for time-dependent changes in Wenckebach patterns, whereas exclusion of facilitation led to consistent overestimation of the degree of AV block at each cycle length. We conclude that a mathematical model incorporating terms to describe recovery, facilitation, and fatigue accurately predicts a wide range of Wenckebach-type behavior and that complex conduction patterns of the AV node can be fully accounted for by simple functional AV nodal properties.


Lamotrigine in the treatment of bipolar disorder.
Lamotrigine is a novel anticonvulsant agent that has recently been introduced as a long-term treatment in bipolar disorder. Its role in the treatment of epilepsy is based on its actions to decrease ion channel conductance and antagonise glutamatergic function. Therefore, it has a mode of action unlike other agents used on a long-term basis in mood disorders. The evidence for efficacy is stronger for the prevention of depressive, rather than manic, episodes. The pivotal trials are in bipolar I disorder, but there is interest in its actions in patients with bipolar II and spectrum conditions. Its efficacy in other psychiatric conditions remains to be properly established. It is well tolerated and, with careful prescribing, the incidence of rash occurs no more than with placebo; however this is still a concern. Although usually well tolerated, headache, insomnia and drowsiness are probably the most common side effects.


Active Smarter Kids (ASK): Rationale and design of a cluster-randomized controlled trial investigating the effects of daily physical activity on children's academic performance and risk factors for non-communicable diseases.
Evidence is emerging from school-based studies that physical activity might favorably affect children's academic performance. However, there is a need for high-quality studies to support this. Therefore, the main objective of the Active Smarter Kids (ASK) study is to investigate the effect of daily physical activity on children's academic performance. Because of the complexity of the relation between physical activity and academic performance it is important to identify mediating and moderating variables such as cognitive function, fitness, adiposity, motor skills and quality of life (QoL). Further, there are global concerns regarding the high prevalence of lifestyle-related non-communicable diseases (NCDs). The best means to address this challenge could be through primary prevention. Physical activity is known to play a key role in preventing a host of NCDs. Therefore, we investigated as a secondary objective the effect of the intervention on risk factors related to NCDs. The purpose of this paper is to describe the design of the ASK study, the ASK intervention as well as the scope and details of the methods we adopted to evaluate the effect of the ASK intervention on 5 (th) grade children. The ASK study is a cluster randomized controlled trial that includes 1145 fifth graders (aged 10 years) from 57 schools (28 intervention schools; 29 control schools) in Sogn and Fjordane County, Norway. This represents 95.3 % of total possible recruitment. Children in all 57 participating schools took part in a curriculum-prescribed physical activity intervention (90 min/week of physical education (PE) and 45 min/week physical activity, in total; 135 min/week). In addition, children from intervention schools also participated in the ASK intervention model (165 min/week), i.e. a total of 300 min/week of physical activity/PE. The ASK study was implemented over 7 months, from November 2014 to June 2015. We assessed academic performance in reading, numeracy and English using Norwegian National tests delivered by The Norwegian Directorate for Education and Training. We assessed physical activity objectively at baseline, midpoint and at the end of the intervention. All other variables were measured at baseline and post-intervention. In addition, we used qualitative methodologies to obtain an in-depth understanding of children's embodied experiences and pedagogical processes taking place during the intervention. If successful, ASK could provide strong evidence of a relation between physical activity and academic performance that could potentially inform the process of learning in elementary schools. Schools might also be identified as effective settings for large scale public health initiatives for the prevention of NCDs. Clinicaltrials.gov ID nr: NCT02132494 . Date of registration, 6(th) of May, 2014.


Theoretical simulation of tumour oxygenation and results from acute and chronic hypoxia.
The tumour microenvironment is considered to be responsible for the outcome of cancer treatment and therefore it is extremely important to characterize and quantify it. Unfortunately, most of the experimental techniques available now are invasive and generally it is not known how this influences the results. Non-invasive methods on the other hand have a geometrical resolution that is not always suited for the modelling of the tumour response. Theoretical simulation of the microenvironment may be an alternative method that can provide quantitative data for accurately describing tumour tissues. This paper presents a computerized model that allows the simulation of the tumour oxygenation. The model simulates numerically the fundamental physical processes of oxygen diffusion and consumption in a two-dimensional geometry in order to study the influence of the different parameters describing the tissue geometry. The paper also presents a novel method to simulate the effects of diffusion-limited (chronic) hypoxia and perfusion-limited (acute) hypoxia. The results show that all the parameters describing tissue vasculature are important for describing tissue oxygenation. Assuming that vascular structure is described by a distribution of inter-vessel distances, both the average and the width of the distribution are needed in order to fully characterize the tissue oxygenation. Incomplete data, such as distributions measured in a non-representative region of the tissue, may not give relevant tissue oxygenation. Theoretical modelling of tumour oxygenation also allows the separation between acutely and chronically hypoxic cells, a distinction that cannot always be seen with other methods. It was observed that the fraction of acutely hypoxic cells depends not only on the fraction of collapsed blood vessels at any particular moment, but also on the distribution of vessels in space as well. All these suggest that theoretical modelling of tissue oxygenation starting from the basic principles is a robust method that can be used to quantify the tissue oxygenation and to provide input parameters for other simulations.


Differentiation between aberrant ventricular conduction and ventricular ectopy in atrial fibrillation using RR interval scattergram.
Differentiation between aberrant ventricular conduction and ventricular ectopy during atrial fibrillation (AF) is of etiologic, prognostic, and therapeutic importance. We developed a noninvasive technique to diagnose aberrant ventricular conduction and ventricular ectopy in AF. We studied the Holter ECGs of 34 patients with paroxysmal AF and 62 patients with chronic AF. In all the patients, frequent wide QRS complexes were observed, and 32 patients were shown by electrophysiological examination to have ventricular ectopies or aberrant ventricular conductions. We obtained the RR interval scattergrams by plotting sequential pairs of RR intervals. Each point has the (n)th RR interval as its x value and the (n + 1)th RR interval as its y value. The irregularity of the RR intervals in AF resulted in widely scattered points delineated by the envelope along the axes. The y value of the envelope along the x axis indicates the shortest coupling interval to the preceding RR interval. Therefore, this curve defines the functional refractory period of atrioventricular conduction. The scattergram of the RR interval pairs immediately preceding the aberrant conduction (coupling points of aberrant conduction) specifically distributed along the envelope. In contrast, the coupling points of ventricular ectopies showed different distributions that had no relation to the envelope. That is, it included three typical patterns, ie, linear distribution below the envelope, linear distribution partially overlapped in the area of normal AF conduction, and chaotic distribution in the AF area. None of the scattergrams of ventricular ectopies showed curvilinear distribution along the envelope as aberrant conduction did. The specific distribution of the aberrant conduction on the RR interval scattergram suggested that aberrant conduction in AF could result from the difference of refractory periods between the AV node and bundle branch block. We conclude that the RR interval scattergram makes it possible to differentiate between aberrant ventricular conduction and ventricular ectopy in atrial fibrillation, and thus, it is a useful noninvasive clinical tool.


Interaction With Social Robots: Improving Gaze Toward Face but Not Necessarily Joint Attention in Children With Autism Spectrum Disorder.
It is widely recognized that robot-based interventions for autism spectrum disorders (ASD) hold promise, but the question remains as to whether social humanoid robots could facilitate joint attention performance in children with ASD. In this study, responsive joint attention was measured under two conditions in which different agents, a human and a robot, initiated joint attention via video. The participants were 15 children with ASD (mean age: 4.96 ± 1.10 years) and 15 typically developing (TD) children (mean age: 4.53 ± 0.90 years). In addition to analyses of fixation time and gaze transitions, a longest common subsequence approach (LCS) was employed to compare participants' eye movements to a predefined logical reference sequence. The fixation of TD toward agent's face was earlier and longer than children with ASD. Moreover, TD showed a greater number of gaze transitions between agent's face and target, and higher LCS scores than children with ASD. Both groups showed more interests in the robot's face, but the robot induced a lower proportion of fixation time on the target. Meanwhile participants showed similar gaze transitions and LCS results in both conditions, suggesting that they could follow the logic of the joint attention task induced by the robot as well as human. We have discussed the implications for the effects and applications of social humanoid robots in joint attention interventions.


Fifty important research questions in microbial ecology.
Microbial ecology provides insights into the ecological and evolutionary dynamics of microbial communities underpinning every ecosystem on Earth. Microbial communities can now be investigated in unprecedented detail, although there is still a wealth of open questions to be tackled. Here we identify 50 research questions of fundamental importance to the science or application of microbial ecology, with the intention of summarising the field and bringing focus to new research avenues. Questions are categorised into seven themes: host-microbiome interactions; health and infectious diseases; human health and food security; microbial ecology in a changing world; environmental processes; functional diversity; and evolutionary processes. Many questions recognise that microbes provide an extraordinary array of functional diversity that can be harnessed to solve real-world problems. Our limited knowledge of spatial and temporal variation in microbial diversity and function is also reflected, as is the need to integrate micro- and macro-ecological concepts, and knowledge derived from studies with humans and other diverse organisms. Although not exhaustive, the questions presented are intended to stimulate discussion and provide focus for researchers, funders and policy makers, informing the future research agenda in microbial ecology.


Father group leaders' experiences of creating an arena for father support - A qualitative study.
Parental classes consisting of only fathers, hereafter referred to as father groups, have existed in Sweden since the mid-1990s. To describe the father group leaders' perspectives on and experiences of father groups. A qualitative study was conducted using content analysis. Eleven individual interviews with father group leaders were conducted. An interview guide was used. The study was set in different parts of Sweden. The participants were eleven men who are father group leaders. They were recruited using the snowball method. The father group leaders described how participants often have high education levels and orderly social circumstances. The leaders described that the groups increased the fathers' reflection about parenthood, which could benefit the whole family in both the short and long term. The father group leaders reported that some of the topics in the sessions directly influenced fathers to discuss issues regarding equality and their co-parenting relationship. They also argued that participating in father groups might help convince fathers to take more parental leave and build stronger relationships with their partner and child.


Rapid evolution of α-gliadin gene family revealed by analyzing Gli-2 locus regions of wild emmer wheat.
α-Gliadins are a major group of gluten proteins in wheat flour that contribute to the end-use properties for food processing and contain major immunogenic epitopes that can cause serious health-related issues including celiac disease (CD). α-Gliadins are also the youngest group of gluten proteins and are encoded by a large gene family. The majority of the gene family members evolved independently in the A, B, and D genomes of different wheat species after their separation from a common ancestral species. To gain insights into the origin and evolution of these complex genes, the genomic regions of the Gli-2 loci encoding α-gliadins were characterized from the tetraploid wild emmer, a progenitor of hexaploid bread wheat that contributed the AABB genomes. Genomic sequences of Gli-2 locus regions for the wild emmer A and B genomes were first reconstructed using the genome sequence scaffolds along with optical genome maps. A total of 24 and 16 α-gliadin genes were identified for the A and B genome regions, respectively. α-Gliadin pseudogene frequencies of 86% for the A genome and 69% for the B genome were primarily caused by C to T substitutions in the highly abundant glutamine codons, resulting in the generation of premature stop codons. Comparison with the homologous regions from the hexaploid wheat cv. Chinese Spring indicated considerable sequence divergence of the two A genomes at the genomic level. In comparison, conserved regions between the two B genomes were identified that included α-gliadin pseudogenes containing shared nested TE insertions. Analyses of the genomic organization and phylogenetic tree reconstruction indicate that although orthologous gene pairs derived from speciation were present, large portions of α-gliadin genes were likely derived from differential gene duplications or deletions after the separation of the homologous wheat genomes ~ 0.5 MYA. The higher number of full-length intact α-gliadin genes in hexaploid wheat than that in wild emmer suggests that human selection through domestication might have an impact on α-gliadin evolution. Our study provides insights into the rapid and dynamic evolution of genomic regions harboring the α-gliadin genes in wheat.


[Theoretic basis on the same therapeutic program for different degenerative brain diseases in terms of the Governor Vessel: Alzheimer's disease and Parkinson's disease].
Through the consultation of TCM ancient classical theory, the relationship of kidney essence, marrow and brain is analyzed. It is discovered that the degenerative brain diseases, represented by Alzheimer's disease (AD) and Parkinson's disease (PD) share the same etiological basis as "kidney essence deficiency and brain marrow emptiness" and have the mutual pathological outcomes as yang qi declining. The Governor Vessel gathers yang qi of the whole body and maintains the normal functional activity of zangfu organs in the human body through the storage, regulation and invigoration of yang qi. It is viewed that the theory of the Governor Vessel is applied to treat the different degenerative brain diseases, which provides the theoretic support and practice guide for the thought of TCM as the same therapeutic program for the different diseases. As a result, the degenerative brain diseases can be retarded and the approach is provided to the effective prevention and treatment of degenerative diseases in central nerve system:


Association between vancomycin day 1 exposure profile and outcomes among patients with methicillin-resistant Staphylococcus aureus infective endocarditis.
Given the critical importance of early appropriate therapy, a retrospective cohort (2002 to 2013) was performed at the Detroit Medical Center to evaluate the association between the day 1 vancomycin exposure profile and outcomes among patients with MRSA infective endocarditis (IE). The day 1 vancomycin area under the concentration-time curve (AUC0-24) and the minimum concentration at 24 h (Cmin 24) was estimated for each patient using the Bayesian procedure in ADAPT 5, an approach shown to accurately predict the vancomycin exposure with low bias and high precision with limited pharmacokinetic sampling. Initial MRSA isolates were collected and vancomycin MIC was determined by broth microdilution (BMD) and Etest. The primary outcome was failure, defined as persistent bacteremia (≥7 days) or 30-day attributable mortality. Classification and regression tree analysis (CART) was used to determine the vancomycin exposure variables associated with an increased probability of failure. In total, 139 patients met study criteria; 76.3% had right-sided IE, 16.5% had left-sided IE, and 7.2% had both left and right-sided IE. A total of 89/139 (64%) experienced failure by composite definition. In the CART analysis, failure was more pronounced in patients with an AUC0-24/MIC as determined by BMD of ≤600 relative to those with AUC0-24/MIC as determined by BMD of >600 (69.8% versus 54.7%, respectively, P = 0.073). In the logistic regression analysis, an AUC/MIC as determined by BMD of ≤600 (adjusted odds ratio, 2.3; 95% confidence interval, 1.01 to 5.37; P = 0.047) was independently associated with failure. Given the retrospective nature of the present study, further prospective studies are required but these data suggest that patients with an AUC0-24/MIC as determined by BMD of ≤600 present an increased risk of failure.


The therapeutic mode of action of 4-aminopyridine in cerebellar ataxia.
Episodic ataxia type 2 (EA2) is a hereditary cerebellar ataxia associated with mutations in the P/Q-type voltage-gated calcium (Ca(2+)) channels. Therapeutic approaches for treatment of EA2 are very limited. Presently, the potassium (K(+)) channel blocker 4-aminopyridine (4-AP) constitutes the most promising treatment, although its mechanism of action is not understood. Here we show that, in contrast to what is commonly believed, therapeutic concentrations of 4-AP do not increase the inhibitory drive of cerebellar Purkinje cells. Instead, 4-AP restores the severely diminished precision of pacemaking in Purkinje cells of EA2 mutant mice by prolonging the action potential and increasing the action potential afterhyperpolarization. Consistent with this mode of action, the therapeutic efficacy of 4-AP was comparable, and not additive, to chlorzoxazone, an activator of Ca(2+)-dependent K(+) channels that also restores the precision of Purkinje cell pacemaking. The likely target of 4-AP at the concentrations used are the K(v)1 family of K(+) channels, possibly the K(v)1.5 subtype. Because at higher concentrations 4-AP blocks a large array of K(+) channels and is a proconvulsant, use of selective K(v)1 channel blockers is likely to be a safer substitute for treatment of cerebellar ataxia.


Hedgehog Signaling Strength Is Orchestrated by the mir-310 Cluster of MicroRNAs in Response to Diet.
Since the discovery of microRNAs (miRNAs) only two decades ago, they have emerged as an essential component of the gene regulatory machinery. miRNAs have seemingly paradoxical features: a single miRNA is able to simultaneously target hundreds of genes, while its presence is mostly dispensable for animal viability under normal conditions. It is known that miRNAs act as stress response factors; however, it remains challenging to determine their relevant targets and the conditions under which they function. To address this challenge, we propose a new workflow for miRNA function analysis, by which we found that the evolutionarily young miRNA family, the mir-310s (mir-310/mir-311/mir-312/mir-313), are important regulators of Drosophila metabolic status. mir-310s-deficient animals have an abnormal diet-dependent expression profile for numerous diet-sensitive components, accumulate fats, and show various physiological defects. We found that the mir-310s simultaneously repress the production of several regulatory factors (Rab23, DHR96, and Ttk) of the evolutionarily conserved Hedgehog (Hh) pathway to sharpen dietary response. As the mir-310s expression is highly dynamic and nutrition sensitive, this signal relay model helps to explain the molecular mechanism governing quick and robust Hh signaling responses to nutritional changes. Additionally, we discovered a new component of the Hh signaling pathway in Drosophila, Rab23, which cell autonomously regulates Hh ligand trafficking in the germline stem cell niche. How organisms adjust to dietary fluctuations to sustain healthy homeostasis is an intriguing research topic. These data are the first to report that miRNAs can act as executives that transduce nutritional signals to an essential signaling pathway. This suggests miRNAs as plausible therapeutic agents that can be used in combination with low calorie and cholesterol diets to manage quick and precise tissue-specific responses to nutritional changes.


Oral glucose augments the counterregulatory hormone response during insulin-induced hypoglycemia in humans.
It has been suggested that the counterregulatory hormone (CRH) response to acute hypoglycemia is triggered via glucose sensors situated in either the hypothalamus or the portohepatic area. If the latter were critical during hypoglycemia, one would anticipate that ingestion of glucose, by raising glucose levels in the portal circulation, should attenuate CRH responses previously described in animal studies. To evaluate the effect of raising portal, but not peripheral, glucose levels during insulin-induced hypoglycemia, we performed hypoglycemic clamp studies in five healthy adult males on two occasions. On one occasion, subjects received oral glucose (OG) (25 g) during hypoglycemia; and on one occasion, noncarbohydrate-containing drink of equal volume, while maintaining plasma glucose at 55 +/- 2 mg/dL (3.08 mmol/L). As a result, there were no significant differences in systemic plasma glucose levels between the two hypoglycemic clamp studies, and basal CRH concentrations were also similar. As expected, there was a brisk rise in all CRH during the control (hypoglycemia+noncarbohydrate drink) study. In the experimental study, administration of OG (hypoglycemia+OG), to raise intraportal glucose levels during systemic hypoglycemia, did not attenuate CRH responses. Indeed, OG enhanced the rise in epinephrine, glucagon, and GH. Increases in cortisol and norepinephrine did not differ between the two studies. Therefore, our data suggest that increasing the level of glucose in the portal vein above that in the systemic circulation, during hypoglycemia, enhances (rather than suppresses) CRH responses. Thus, ingestion of glucose may reverse hypoglycemia directly by provision of substrate, as well as indirectly by stimulating counteregulatory mechanisms.


Brain systems mediating semantic and syntactic processing in deaf native signers: biological invariance and modality specificity.
Studies of written and spoken language suggest that nonidentical brain networks support semantic and syntactic processing. Event-related brain potential (ERP) studies of spoken and written languages show that semantic anomalies elicit a posterior bilateral N400, whereas syntactic anomalies elicit a left anterior negativity, followed by a broadly distributed late positivity. The present study assessed whether these ERP indicators index the activity of language systems specific for the processing of aural-oral language or if they index neural systems underlying any natural language, including sign language. The syntax of a signed language is mediated through space. Thus the question arises of whether the comprehension of a signed language requires neural systems specific for this kind of code. Deaf native users of American Sign Language (ASL) were presented signed sentences that were either correct or that contained either a semantic or a syntactic error (1 of 2 types of verb agreement errors). ASL sentences were presented at the natural rate of signing, while the electroencephalogram was recorded. As predicted on the basis of earlier studies, an N400 was elicited by semantic violations. In addition, signed syntactic violations elicited an early frontal negativity and a later posterior positivity. Crucially, the distribution of the anterior negativity varied as a function of the type of syntactic violation, suggesting a unique involvement of spatial processing in signed syntax. Together, these findings suggest that biological constraints and experience shape the development of neural systems important for language.


The colorectal cancer disease-specific transcriptome may facilitate the discovery of more biologically and clinically relevant information.
To date, there are no clinically reliable predictive markers of response to the current treatment regimens for advanced colorectal cancer. The aim of the current study was to compare and assess the power of transcriptional profiling using a generic microarray and a disease-specific transcriptome-based microarray. We also examined the biological and clinical relevance of the disease-specific transcriptome. DNA microarray profiling was carried out on isogenic sensitive and 5-FU-resistant HCT116 colorectal cancer cell lines using the Affymetrix HG-U133 Plus2.0 array and the Almac Diagnostics Colorectal cancer disease specific Research tool. In addition, DNA microarray profiling was also carried out on pre-treatment metastatic colorectal cancer biopsies using the colorectal cancer disease specific Research tool. The two microarray platforms were compared based on detection of probesets and biological information. The results demonstrated that the disease-specific transcriptome-based microarray was able to out-perform the generic genomic-based microarray on a number of levels including detection of transcripts and pathway analysis. In addition, the disease-specific microarray contains a high percentage of antisense transcripts and further analysis demonstrated that a number of these exist in sense:antisense pairs. Comparison between cell line models and metastatic CRC patient biopsies further demonstrated that a number of the identified sense:antisense pairs were also detected in CRC patient biopsies, suggesting potential clinical relevance. Analysis from our in vitro and clinical experiments has demonstrated that many transcripts exist in sense:antisense pairs including IGF2BP2, which may have a direct regulatory function in the context of colorectal cancer. While the functional relevance of the antisense transcripts has been established by many studies, their functional role is currently unclear; however, the numbers that have been detected by the disease-specific microarray would suggest that they may be important regulatory transcripts. This study has demonstrated the power of a disease-specific transcriptome-based approach and highlighted the potential novel biologically and clinically relevant information that is gained when using such a methodology.


Pivotal study of iodine I 131 tositumomab for chemotherapy-refractory low-grade or transformed low-grade B-cell non-Hodgkin's lymphomas.
To evaluate the efficacy and safety of tositumomab and iodine I 131 tositumomab (Bexxar; Corixa Corp, Seattle, WA, and GlaxoSmithKline, Philadelphia, PA) in patients with chemotherapy-refractory low-grade or transformed low-grade non-Hodgkin's lymphoma (NHL) and to compare its efficacy to the patients' last qualifying chemotherapy (LQC) regimens. Sixty patients who had been treated with at least two protocol-specified qualifying chemotherapy regimens and had not responded or progressed within 6 months after their LQC were treated with a single course of iodine I 131 tositumomab. Patients had received a median of four prior chemotherapy regimens. A partial or complete response (CR) was observed in 39 patients (65%) after iodine I 131 tositumomab, compared with 17 patients (28%) after their LQC (P <.001). The median duration of response (MDR) was 6.5 months after iodine I 131 tositumomab, compared with 3.4 months after the LQC (P <.001). Two patients (3%) had a CR after their LQC, compared with 12 (20%) after iodine I 131 tositumomab (P <.001). The MDR for CR was 6.1 months after the LQC and had not been reached with follow-up of more than 47 months after iodine I 131 tositumomab. An independent review panel verified that 32 (74%) of the 43 patients with nonequivalent durations of response (> 30 days difference) had a longer duration of response after iodine I 131 tositumomab (P <.001). Only one patient was hospitalized for neutropenic fever. Five patients (8%) developed human antimurine antibodies, and one (2%) developed an elevated TSH level after treatment. Myelodysplasia was diagnosed in four patients in follow-up. A single course of iodine I 131 tositumomab was significantly more efficacious than the LQC received by extensively pretreated patients with chemotherapy-refractory, low-grade, or transformed low-grade NHL and had an acceptable safety profile.


Antimicrobial resistance in 11 hospitals in Puerto Rico: results of an antimicrobial resistance management (ARM) program.
The Antimicrobial Resistance Management Program (ARMP) was established in 1997 at the University of Florida as an ongoing project to document trends in antimicrobial susceptibility patterns in inpatient/outpatient isolates and track resistance that may occur with specific antibiotic use. Institutions are enrolled at no charge and provide a minimum of 3 years of antibiogram/sensitivity report data, which are reviewed to create a customized analysis of antimicrobial susceptibility trends benchmarked against national/regional comparators. The data, in a HIPAA-compliant non-identifying format, comprise a national aggregate database of 28.4 million isolates from 358 institutions. This database was interrogated to determine resistance rates for eleven hospitals in Puerto Rico from 1998-2003 and, as comparators, those in the database from the State of Florida and all U.S. institutions. Between 1996-2003, data on 328,837 isolates collected from 11 hospitals throughout Puerto Rico, 5,388,897 isolates from 46 institutions in Florida, and 24,951,098 isolates from 358 U.S. institutions for the following organisms (number of antibiotics tested against) were reviewed for susceptibility: coagulase-negative staphylococci (14)/Staphylococcus epidermidis (18), Enterococcus faecalis (7), Enterococcus faecium (5), Enterococcus species (4), Escherichia coli (24), Klebsiella pneumoniae (24), Proteus mirabilis (22), Pseudomonas aeruginosa (14), Serratia marcescens (22), Staphylococcus aureus (23), and Streptococcus pneumoniae (9). Antimicrobial resistance in Puerto Rico varied organism to organism from that observed in Florida and nationally. This first broad analysis of antimicrobial resistance in Puerto Rico provides important baseline data, both for sentinel surveillance programs and for determining strategies for intervention.


Actin-based motility of intracellular pathogens.
The actin cytoskeleton is harnessed by several pathogenic bacteria that are capable of entering into non-phagocytic cells, the so-called 'invasive bacteria'. Among them, a few also exploit the host actin cytoskeleton to move intra- and inter-cellularly. Our knowledge of the basic mechanisms underlying actin-based motility has dramatically increased and the list of bacteria that are able to move in this way is also increasing including not only Listeria, Shigella and Rickettsia species but also Mycobacterium marinum and Burkholderia pseudomallei. In all cases the central player is the Arp2/3 complex. Vaccinia virus moves intracellularly on microtubules and just after budding, triggers actin polymerization and the formation of protrusions similar to that of adherent enteropathogenic Escherichia coli, that involve the Arp2/3 complex and facilitate its inter-cellular spread.


Serum melatonin levels and antioxidant capacities after consumption of pineapple, orange, or banana by healthy male volunteers.
Melatonin is a naturally occurring molecule biosynthesized by the pineal gland of vertebrates; it also has been identified in many plants. It is considered an important antioxidant and may retard the development of some neurodegenerative diseases and cancer. Previous studies in humans have measured melatonin metabolites in urine and have indicated that melatonin-containing foods may provide dietary melatonin. This study tested whether the consumption of fruits or fruit juice containing melatonin would influence the serum melatonin concentration and antioxidant status. In this crossover study, 12 healthy male volunteers took either juice extracted from one kilogram of orange or pineapple or two whole bananas, with a 1-wk washout period between the fruit or fruit juices. An enzyme-linked immunosorbent (ELISA) assay was used to determine the serum melatonin concentration. Serum antioxidant capacity was determined by ferric reducing antioxidant power (FRAP) assay and oxygen radical antioxidant capacity (ORAC) assay. The highest serum melatonin concentration was observed at 120 min after fruit consumption, and compared with before consumption levels, their values were significantly increased for pineapple (146 versus 48 pg/mL P = 0.002), orange (151 versus 40 pg/mL, P = 0.005), and banana (140 versus 32 pg/mL, P = 0.008), respectively. Serum antioxidant capacity following fruit consumption also significantly increased in both the FRAP (7-14% increase, P ≤ 0.004) and ORAC (6-9% increase, P = 0.002) assays. Both the serum FRAP and ORAC values strongly correlated with serum melatonin concentration for all three fruits. These findings suggest that tropical fruit consumption increases the serum melatonin concentrations and also raises the antioxidant capacity in the serum of healthy volunteers in proportion to serum melatonin levels.


The relationship between SCHIP enrollment and hospitalizations for ambulatory care sensitive conditions in California.
The State Children's Health Insurance Program (SCHIP) was implemented in 1998, providing new funds for states to cover uninsured children. This study examines the relationship between SCHIP implementation in California and hospitalizations for ambulatory care sensitive conditions (ACSCs), an indicator of primary care access and quality. We use administrative SCHIP enrollment records for urban California counties, linked with corresponding rates of hospitalization for seven ACSCs among children ages 1-18 for 1996-2000. Results from multivariate regression models indicate that increases of 1 percentage point in SCHIP enrollment are associated with reductions of 0.42 ACSC admissions per 100,000 children age 1-18 (p = 0.009). Models that use lagged effects of SCHIP enrollment indicate an even stronger relationship. These are population-level relationships, and translate to much larger effects on the specific population subset that enrolled in SCHIP. These results suggest a strong beneficial effect of SCHIP on primary care among the children covered.


Subjective and physiological effects after controlled Sativex and oral THC administration.
Sativex is a cannabis-plant extract delivering nearly 1:1 Δ(9)-tetrahydrocannabinol (THC) and cannabidiol (CBD) by oromucosal spray. It has been suggested that CBD attenuates THC-induced tachycardia, anxiety, and euphoria. In this study, pharmacodynamic effects were compared over 10.5 h in nine cannabis smokers randomly assigned to receive placebo, 5 and 15 mg oral synthetic THC, and low (5.4 mg THC, 5.0 mg CBD) and high (16.2 mg THC, 15.0 mg CBD) doses of Sativex. At therapeutic doses, no substantial CBD-induced modulation of THC's effects was evident. Oral THC and Sativex produced similar, clinically insignificant increases in heart rate, anxiety, and "good drug effects" with no serious adverse events. Oral and oromucosal THC have slower absorption, lower rate of THC delivery to the brain, and fewer associated adverse events as compared with smoked cannabis. These results indicate that Sativex has a pharmacodynamic safety profile comparable to that of oral THC at low, therapeutic doses.


The kaiser permanente northwest cardiovascular risk factor management program: a model for all.
Proof of the effectiveness of preventive measures that reduce established risk traits for atherothrombotic disorders has spurred attempts to systematically apply these interventions among susceptible populations. One such attempt is the Cardiovascular Risk Factor Management (CVRFM) Program, launched in 2003 to optimize clinical management and outcomes for 75,000 Kaiser Permanente Northwest Region (KPNW) members with atherosclerotic cardiovascular disease (CVD) or hypertension. The CVRFM Program is a centralized, multidisciplinary, proactive telephone-based clinical management intervention consisting of an "outreach" call, an interview, a mailed individualized care plan and information packet, regular follow-up (including protocolized medication management) and-when "goal status" is achieved-transfer of the patient to a maintenance plan.Quarterly evaluation of effectiveness entailed measurement of a range of clinical, utilization, and member satisfaction outcomes. Results by the fourth quarter were outstanding: For example, >98% of participants with coronary disease or diabetes had LDL cholesterol testing, >90% of coronary patients received aspirin or statin treatment, 99% were "extremely" or "very" satisfied with the program, and reductions were observed in the number of hospitalizations and visits to the emergency department and clinic. Mathematical models predict a decrease in myocardial infarctions and cardiovascular mortality within two years after implementing the program, the underlying principles of which should yield similar improvement in other Kaiser Permanente (KP) Regions and in other health care organizations.


Promoter methylation of E-cadherin, p16, and RAR-β(2) genes in breast tumors and dietary intake of nutrients important in one-carbon metabolism.
Aberrant DNA methylation plays a critical role in carcinogenesis, and the availability of dietary factors involved in 1-carbon metabolism may contribute to aberrant DNA methylation. We investigated the association of intake of folate, vitamins B(2), B(6), B(12), and methionine with promoter methylation of E-cadherin, p16, and RAR-β(2) genes in archived tumor tissues from incident, primary breast cancer cases in a population-based case-control study. Real-time methylation-specific PCR was performed on 803 paraffin-embedded samples; usual dietary intake was queried from a food frequency questionnaire. Unconditional logistic regression was used to derive adjusted odds ratios and 95% confidence intervals for likelihood of promoter methylation for high compared to low intake of those 1-carbon nutrients. Overall, in case-case comparisons, dietary intakes of folate, vitamins B(2), B(6), B(12), and methionine were not associated with likelihood of promoter methylation of E- cadherin, p16, and RAR-β(2) for all cases combined or within strata defined by menopausal status and estrogen receptor status in this study. This finding, however, does not exclude the possibility that intake of such nutrients might have the ability to modulate promoter methylation in normal or premalignant (dysplastic) breast tissue.


Two-stage regression of high-density scalp electroencephalograms visualizes force regulation signaling during muscle contraction.
A critical feature for the maintenance of precise skeletal muscle force production by the human brain is its ability to configure motor function activity dynamically and adaptively in response to visual and somatosensory information. Existing studies have concluded that not only the sensorimotor area but also distributed cortical areas act cooperatively in the generation of motor commands for voluntary force production to the desired level. However, less attention has been paid to such physiological mechanisms in conventional brain-computer interface (BCI) design and implementation. We proposed a new, physiologically inspired two-stage decoding method to see its contribution on accuracy improvement of BCI. We performed whole-head high-density scalp electroencephalographic (EEG) recording during a right finger force-matching task at three strength levels (20%, 40%, and 60% maximal voluntary contraction following a resting state). A two-stage regression approach was employed that decodes muscle contraction level from EEG signals in the multi-level force-matching task and translates them into: (1) presence/absence of muscle contraction as a first stage; and (2) muscle contraction level as a second stage. Dimensionality reduction of the EEG signals, using principal component analysis, avoided multicollinearity during multiple regression, and data-driven stepwise multiple regression identified EEG components that were involved in the multi-level force-matching task. An alternatively tuned two-stage regressor accurately decoded muscle contraction level with online processing rather than the conventional decoders, and identified EEG components that were related to voluntary force production. Relaxation/contraction state-dependent EEG components were localized dominantly in the contralateral parieto-temporal regions, whereas multi-level force regulation-dependent EEG components came from the fronto-parietal regions. Our findings identify respective cortical signalings during relaxation/contraction and multi-level force regulation using a sensor-based approach with EEG. Simulation-based assessment of the current physiologically inspired decoding technique proved improved accuracy in online BCI control.


Geography of follicle formation in the embryonic mouse ovary impacts activation pattern during the first wave of folliculogenesis.
During embryonic development, mouse female germ cells enter meiosis in an anterior-to-posterior wave believed to be driven by retinoic acid. It has been proposed that ovarian follicle formation and activation follow the same general wave of meiotic progression; however, the precise anatomic specification of these processes has not been delineated. Here, we created a mouse line using Mvh, Gdf9, and Zp3 promoters to drive distinct temporal expression of three fluorescent proteins in the oocytes and to identify where the first follicle cohort develops. The fluorescent profile revealed that the first growing follicles consistently appeared in a specific region of the ovary, the anterior-dorsal region, which led us to analyze if meiotic onset occurred earlier in the dorsal ovarian region. Surprisingly, in addition to the anterior-to-posterior wave, we observed an early meiotic entry in the ventral region of the ovary. This additional anatomic stratification of meiosis contrasts with the localization of the initial follicle formation and activation in the dorsal region of the ovary. Therefore, our study suggests that the specification of cortical and medullar areas in the ventral and dorsal regions on the ovary, rather than the onset of meiosis, impacts where the first follicle activation event occurs.


Preclinical biological and physicochemical evaluation of two-photon engineered 3D biomimetic copolymer scaffolds for bone healing.
A major challenge in orthopedics is the repair of large non-union bone fractures. A promising therapy for this indication is the use of biodegradable bioinspired biomaterials that stabilize the fracture site, relieve pain and initiate bone formation and healing. This study uses a multidisciplinary evaluation strategy to assess immunogenicity, allergenicity, bone responses and physicochemical properties of a novel biomaterial scaffold. Two-photon stereolithography generated personalized custom-built scaffolds with a repeating 3D structure of Schwarz Primitive minimal surface unit cell with a specific pore size of ∼400 μm from three different methacrylated poly(d,l-lactide-co-ε-caprolactone) copolymers with lactide to caprolactone monomer ratios of 16 : 4, 18 : 2 and 9 : 1. Using in vitro and in vivo assays for bone responses, immunological reactions and degradation dynamics, we found that copolymer composition influenced the scaffold physicochemical and biological properties. The scaffolds with the fastest degradation rate correlated with adverse cellular effects and mechanical stiffness correlated with in vitro osteoblast mineralization. The physicochemical properties also correlated with in vivo bone healing and immune responses. Overall these observations provide compelling support for these scaffolds for bone repair and illustrate the effectiveness of a promising multidisciplinary strategy with great potential for the preclinical evaluation of biomaterials.


Genetic and functional evidence for a role for SLC11A1 in susceptibility to otitis media in early childhood in a Western Australian population.
Otitis media (OM) is a common disease in early childhood characterised by inflammation of the middle ear. Susceptibility to recurrent acute OM (rAOM; ≥3 episodes AOM in 6 months) and chronic OM with effusion (COME; middle ear effusion ≥3 months) is 40-70% heritable. Three bacterial pathogens commonly associated with OM, Streptococcus pneumoniae (Sp), non-typeable Haemophilus influenzae (NTHi) and Moraxella catarrhalis (Mc), have been observed within adenoids and as facultative intracellular pathogens that invade and survive in mononuclear cells. Case/pseudo-control conditional logistic regression analysis of variants in the SLC11A1 gene, initially identified for its role in resistance to intra-macrophage pathogens in mice, revealed association with OM at four polymorphisms (Pbest=0.025) in 531 families (660 affected children) from the Western Australian Family Study of Otitis Media. This included association at the functional promoter GTn polymorphism (rs34448891) with alleles that regulate high (allele 3; odds ratio=1.2, 95% CI 1.00-1.44, P=0.04) versus low (allele 2; odds ratio=0.83, 95% CI 0.69-0.99, P=0.04) SLC11A1 expression. Haplotype and stepwise conditional logistic regression analyses support a single genetic effect in the proximal region of SLC11A1, with the haplotype 3_C_C_G across rs34448891_rs2276631_rs3731865_rs2695343 significantly (P=0.008) over-transmitted to affected offspring. Stratified analysis showed no association with OM in children who had undergone adenoidectomy (296 children), whereas children with adenoids intact (364 children) showed improved significance at the GTn polymorphism (allele 3: odds ratio=1.38, 95% CI=1.10-1.75, P=0.006). Quantitative RT/PCR demonstrated high expression of SLC11A1 in mononuclear cells isolated from adenoid tissue, with a trend for decreased expression with increasing copies of GTn allele 2. Expression of SLC11A1 was enhanced at 12 (P=1.2×10(-3)) and 24h (P<1.0×10(-4)) after infection of Mono-Mac-6 cells with NTHi. This study identifies SLC11A1 as a novel candidate for OM susceptibility, particularly in children with adenoids intact. Further analysis in other cohorts is required to validate these observations.


Coumestrol from the national cancer Institute's natural product library is a novel inhibitor of protein kinase CK2.
Casein kinase 2 (CK2) is involved in various cellular events such as proliferation, apoptosis, and the cell cycle. CK2 overexpression is associated with multiple human cancers and may therefore be a promising target for cancer therapy. To identity novel classes of inhibitors for CK2, we screened a natural product library obtained from National Cancer Institute. The quantitative luminescent kinase assay ADP-Glo™ was used to screen CK2 inhibitors from the natural product library. The same assay was used to determine cell-free dose-dependent response of CK2 inhibitors and conduct a kinetic study. Docking was performed to predict the binding patterns of selected CK2 inhibitors. Western blot analysis was used to evaluate Akt phosphorylation specific to CK2 and apoptosis effect. The cell viability assay CellTiter-Glo(®) was used to evaluate the inhibition effects of CK2 inhibitors on cancer cells. We identified coumestrol as a novel reversible ATP competitive CK2 inhibitor with an IC(50) value of 228 nM. Coumestrol is a plant-derived compound that belongs to the class of phytoestrogens, natural compounds that mimic the biological activity of estrogens. In our study, coumestrol showed high selectivity among 13 kinases. The hydrogen bonds formed between coumestrol and the amino acids in the ATP binding site were first reviewed by a molecular docking study that suggested a possible interaction of coumestrol with the hinge region of ATP site of CK2. In addition, coumestrol inhibited cancer cell growth partially through down-regulation of CK2-specific Akt phosphorylation. Finally, coumestrol exerted strong inhibition effects on the growth of three cancer cell lines. Our study shows that coumestrol, a novel ATP competitive and cell permeable CK2 inhibitor with submicromolar IC(50), had inhibition effects on the growth of three cancer cell lines and may represent a promising class of CK2 inhibitors.


Nitrate reduction and the growth of Veillonella alcalescens.
Veillonella alcalescens, a strict anaerobe, was found to possess a nitrate reductase system which has characteristics of both assimilatory and respiratory nitrate reduction. The nitrate reductase has been identified tentatively as a particulate enzyme which utilizes a variety of electron donors for the reduction of nitrate. By use of (15)N-labeled nitrate, it was shown that under appropriate conditions nitrate nitrogen is incorporated into cell material. V. alcalescens grown on pyruvate and nitrate has a greater growth rate than cells grown on pyruvate alone. Growth can occur in a medium with hydrogen and nitrate as the sole energy source. Ammonium chloride decreases the rate of nitrate reduction but does not completely inhibit reduction or incorporation. The results suggest that nitrate assimilation and respiration are not as distinct as in some other organisms.


Tuning the Exchange Bias on a Single Atom from 1 mT to 10 T.
Shrinking spintronic devices to the nanoscale ultimately requires localized control of individual atomic magnetic moments. At these length scales, the exchange interaction plays important roles, such as in the stabilization of spin-quantization axes, the production of spin frustration, and creation of magnetic ordering. Here, we demonstrate the precise control of the exchange bias experienced by a single atom on a surface, covering an energy range of 4 orders of magnitude. The exchange interaction is continuously tunable from milli-eV to micro-eV by adjusting the separation between a spin-1/2 atom on a surface and the magnetic tip of a scanning tunneling microscope. We seamlessly combine inelastic electron tunneling spectroscopy and electron spin resonance to map out the different energy scales. This control of exchange bias over a wide span of energies provides versatile control of spin states, with applications ranging from precise tuning of quantum state properties, to strong exchange bias for local spin doping. In addition, we show that a time-varying exchange interaction generates a localized ac magnetic field that resonantly drives the surface spin. The static and dynamic control of the exchange interaction at the atomic scale provides a new tool to tune the quantum states of coupled-spin systems.


Co-designing obesity prevention interventions together with children: intervention mapping meets youth-led participatory action research.
Youth-led Participatory Action Research (YPAR) involves children throughout the process of developing and implementing interventions. Combining YPAR with a structural approach for designing and planning interventions, such as Intervention Mapping (IM), may further improve implementation and effectiveness of interventions. This paper describes how YPAR and IM were combined in the Kids in Action study. The Kids in Action study aims to improve health behaviors of 9-12-year old children living in a low socioeconomic neighborhood in Amsterdam, by co-designing interventions with these children. At each of four schools 6-8 children (N = 18-24 total per year) and two academic researchers formed participatory groups that met weekly or every fortnight during two school years. An IM expert panel advised the participatory groups on the application of IM. Following the IM protocol, we conducted a participatory needs assessment with children, parents and professionals, in IM-step 1. In IM-step 2, the IM expert panel constructed matrices of program objectives, and the children provided feedback. In collaboration with children programs were designed and produced using an iterative process during IM-steps 3-4. In IM-step 5, the participatory groups and professional community partners designed the implementation plan. Finally, in IM-step 6, the protocol of the process and effect evaluation - executed by academic researchers with input from children - was developed. By combining YPAR and IM, several interventions have been developed and implemented, varying from a school water policy to extracurricular sports activities. Sharing responsibility with children was challenging when combining IM with YPAR. In YPAR children are given as much autonomy as possible, while traditional IM development work is primarily done by academic researchers. Strengths in combining IM and YPAR include the involvement of the end-users - children - throughout the process while at the same time developing interventions based on existing evidence. Time-management, a multidisciplinary team, and flexibility are important conditions when combining IM with YPAR. A strong community project group, with professionals who were willing to help children develop and execute their ideas, was an important success factor. This study can serve as an example to other YPAR studies developing interventions using the IM protocol.


Reactivity of Boronic Acids toward Catechols in Aqueous Solution.
Fundamental information on the reactivities of boronic acids toward catechols in aqueous solution is required in all the fields dealing with boronic acid. However, comprehensive studies on reactivity are often hindered by so-called "proton ambiguity," which makes it impossible for the rate constants of boronic acid and boronate ion to be determined separately. Herein, we set up two reaction systems without proton ambiguity: (1) Alizarin Red S and (2) Tiron with several boronic acids (RB(OH)2) with different pKas and performed kinetic and equilibrium studies on the reaction systems. It was shown that the logarithms of the rate constants of RB(OH)2 and its conjugate boronate ion (RB(OH)3-) decreased and increased linearly, respectively, with increasing pKa of RB(OH)2 for both systems. Consequently, the reactivities of RB(OH)2 and RB(OH)3- were reversed at high RB(OH)2 pKa. It was also shown that the bulky o- substituents of phenylboronic acids retarded the backward reactions, resulting in enhancement of the formation constants of boronic acid-catechol esters.


Degenerative mitral valve regurgitation: best practice revolution.
Degenerative mitral valve disease often leads to leaflet prolapse due to chordal elongation or rupture, and resulting in mitral valve regurgitation. Guideline referral for surgical intervention centres primarily on symptoms and ventricular dysfunction. The recommended treatment for degenerative mitral valve disease is mitral valve reconstruction, as opposed to valve replacement with a bioprosthetic or mechanical valve, because valve repair is associated with improved event free survival. Recent studies have documented a significant number of patients are not referred in a timely fashion according to established guidelines, and when they are subjected to surgery, an alarming number of patients continue to undergo mitral valve replacement. The debate around appropriate timing of intervention for asymptomatic severe mitral valve regurgitation has put additional emphasis on targeted surgeon referral and the need to ensure a very high rate of mitral valve repair, particularly in the non-elderly population. Current clinical practice remains suboptimal for many patients, and this review explores the need for a 'best practice revolution' in the field of degenerative mitral valve regurgitation.


Regulation of insulin-like growth factor-I in skeletal muscle and muscle cells.
Growth hormone (GH) and insulin-like growth factor-I (IGF-I) are potent regulators of muscle mass. Transgenic mice that over-express these proteins exhibit dramatically enlarged skeletal muscles. In contrast, malnutrition, critical illness, sepsis, and aging are all associated with a dramatic reduction in muscle mass and function. The circulating concentration of IGF-I and the expression of IGF-I in skeletal muscle are also reduced during catabolic states. Consequently, GH has been used clinically to increase lean body mass in patients with muscle wasting. Likewise, delivery of IGF-I specifically into muscle has been proposed as a genetic therapy for muscle disorders. A better understanding of the regulation of IGF-I expression in skeletal muscle and muscle cells is therefore of importance. Yet, our knowledge in this area has been limited by a lack of GH responsive muscle cells. In addition the IGF-I gene spans over 90 kb of genomic DNA and it exhibits a very complex regulatory pattern. This review will summarize our knowledge of the control of muscle mass by GH, IGF-I, anabolic steroids, exercise and other growth enhancing hormones. We will also highlight recent advances in the regulation of IGF-I and signal transducers and activators of transcription (Stats) by GH. A special emphasis will be placed on the interaction of IGF-I and proinflammatory cytokines in skeletal muscle and muscle cells.


Coupling of dopamine D1 recognition sites with adenylate cyclase in nuclei accumbens and caudatus of schizophrenics.
Sodium fluoride, guanylimidodiphosphate, and the D1 dopamine receptor agonist SKF 38393 elicited a greater activation of adenylate cyclase in homogenates of caudate nucleus in schizophrenic than in nonschizophrenic subjects used as controls. Similarly, a greater activation of adenylate cyclase by sodium fluoride was observed in the nucleus accumbens of schizophrenics. These findings suggest that the coupling of dopamine D1 recognition sites with adenylate cyclase is more efficient in the brain of the schizophrenic, presumably because of an increased affinity of the G/F protein for guanosine 5'-triphosphate.