An individualized recruitment maneuver for mechanically ventilated patients after cardiac surgery.
The recruitment maneuver (RM) has been shown to improve oxygenation for post-cardiopulmonary bypass (CPB) patients; however, sustained inflation of the lung gives rise to hypotension. The primary goal of our study was to evaluate the safety and efficacy of our proposed RM, defined on the basis of dynamic lung compliance (Cdyn). Twenty-eight patients undergoing elective cardiac surgery with CPB were assigned to two treatment groups: an individualized RM group, in which a pressure equal to 15 ml x real body weight/Cdyn + positive end-expiratory pressure (PEEP) cmH2O was applied for 15 s; and a control RM group, in which a pressure of 20 cmH2O was applied for 25 s. Arterial blood pressure, cardiac output, pulmonary artery pressure, and heart rate (HR) were monitored. Tidal volume (V(T)), and airway pressure were continuously obtained from an expiratory flow meter and pressure monitor. Blood samples were obtained and analyzed with a blood gas analyzer. The changes in HR, mean arterial pressure, mean pulmonary artery pressure, and cardiac index at the end of the RM were not significantly different between the two groups. The mean airway pressure of sustained inflation was 28.3 +/- 1.3 cmH2O in the individualized RM group. The individualized RM significantly improved the Cdyn and partial pressure arterial oxygen/inspiratory fraction of oxygen (P/F) ratio compared with values in the control RM group (P = 0.026 and P = 0.012, respectively). The present study indicates that the individualized RM resulted in minimum changes of hemodynamics and brought about improvement in oxygenation and lung compliance.


Controlled MOCVD growth of Bi2Se3 topological insulator nanoribbons.
Topological insulators are a new class of materials that support topologically protected electronic surface states. Potential applications of the surface states in low dissipation electronic devices have motivated efforts to create nanoscale samples with large surface-to-volume ratios and highly controlled stoichiometry. Se vacancies in Bi(2)Se(3) give rise to bulk conduction, which masks the transport properties of the surface states. We have therefore developed a new route for the synthesis of topological insulator nanostructures using metalorganic chemical vapour deposition (MOCVD). MOCVD allows control of the Se/Bi flux ratio during growth. With the aim of rational growth, we vary the Se/Bi flux ratio, growth time, and substrate temperature, and observe morphological changes which indicate a growth regime in which nanoribbon formation is limited by the Bi precursor mass flow. MOCVD growth of Bi(2)Se(3) nanostructures occurs via a distinct growth mechanism that is nucleated by gold nanoparticles at the base of the nanoribbon. By tuning the reaction conditions, we obtain either single-crystalline ribbons up to 10 μm long or thin micron-sized platelets.


Promoting Personal Recovery in People with Persisting Psychotic Disorders: Development and Pilot Study of a Novel Digital Intervention.
For people with persisting psychotic disorders, personal recovery has become an important target of mental health services worldwide. Strongly influenced by mental health service consumer perspectives, personal recovery refers to being able to live a satisfying and contributing life irrespective of ongoing symptoms and disability. Contact with peers with shared lived experience is often cited as facilitative of recovery. We aimed to develop and pilot a novel recovery-based digitally supported intervention for people with a psychotic illness. We developed a website to be used on a tablet computer by mental health workers to structure therapeutic discussions about personal recovery. Central to the site was a series of video interviews of people with lived experience of psychosis discussing how they had navigated issues within their own recovery based on the Connectedness-Hope-Identity-Meaning-Empowerment model of recovery. We examined the feasibility and acceptability of an 8-session low intensity intervention using this site in 10 participants with persisting psychotic disorders and conducted a proof-of-concept analysis of outcomes. All 10 participants completed the full course of sessions, and it was possible to integrate use of the website into nearly all sessions. Participant feedback confirmed that use of the website was a feasible and acceptable way of working. All participants stated that they would recommend the intervention to others. Post-intervention, personal recovery measured by the Questionnaire for the Process of Recovery had improved by an average standardized effect of d = 0.46, 95% CI [0.07, 0.84], and 8 of the 10 participants reported that their mental health had improved since taking part in the intervention. In-session use of digital resources featuring peer accounts of recovery is feasible and acceptable and shows promising outcomes. A randomized controlled trial is the next step in evaluating the efficacy of this low intensity intervention when delivered in conjunction with routine mental health care.


Chronic allergic fungal sinusitis invading the skull base in an immunocompetent male: illustrative case.
Allergic fungal sinusitis (AFS) is an immunoglobulin E-mediated reaction to fungal organisms in the sinonasal region and can be categorized as acute or chronic. Acute infection is typical in immunocompromised patients, while chronic infection is classically seen in immunocompetent patients. Spread of infection to the skull base is a rare and potentially lethal complication of prolonged infection. Surgical management is frequently augmented with steroid therapy to prevent recurrence. The authors present a case of a 20-year-old African American male with prolonged headaches and blurred vision who was diagnosed with chronic invasive fungal sinusitis resulting in invasion of fungal burden into the anterior skull base and the posterior aspect of the clivus, in addition to complete obliteration of the maxillary sinus. The patient was managed surgically without complication and with gradual improvement in vision. Early management and detection of AFS should be a focus to prevent erosion of the fungal burden into the skull base. Neurosurgery and ear, nose, and throat surgery have a multidisciplinary role in the management of advanced AFS cases.


Pressurized intraperitoneal aerosol chemotherapy (PIPAC) in combination with standard of care chemotherapy in primarily untreated chemo naïve upper gi-adenocarcinomas with peritoneal seeding - a phase II/III trial of the AIO/CAOGI/ACO.
Peritoneal metastasis is a common and dismal evolution of several gastrointestinal (GI) tumors, including gastric, colorectal, hepatobiliary, pancreatic, and other cancers. The therapy of peritoneal metastasis is largely palliative; with the aim of prolonging life and preserving its quality. In the meantime, a significant pharmacological advantage of intraperitoneal chemotherapy was documented in the preclinical model, and numerous clinical studies have delivered promising clinical results. This is a prospective, open, randomized multicenter phase III clinical study with two arms that aims to evaluate the effects of pressurized intraperitoneal aerosol chemotherapy (PIPAC) combined with systemic chemotherapy vs. intravenous systemic chemotherapy alone on patients with metastatic upper GI tumors with a peritoneal seeding. Upper GI-adenocarcinomas originated from biliary tract, pancreas and stomach, or esophago- gastric junction are eligible. Patients in the study are treated with standard of care systemic palliative chemotherapy (mFOLFOX6) vs. PIPAC with intravenous (i.v.) chemotherapy (mFOLFOX6). Patients in first line with first diagnosed peritoneal seeding are eligible. Primary outcome is progression free survival (PFS). PIPAC-procedure is explicit a palliative method but it delivers cytotoxic therapy like in hyperthermic intraperitoneal chemotherapy (HIPEC)-procedure directly to the tumor in a minimally invasive technique, without the need for consideration of the peritoneal-plasma barrier. The technique of PIPAC is minimally invasive and very gentle and the complete procedure takes only round about 45 min and, therefore, optimal in a clearly palliative situation where cure is not the goal. It is also ideal for using this approach in a first line situation, where deepest response should be achieved. The symbiosis of systemic therapy and potentially effective surgery has to be well-planned without deterioration of the patient due to aggressive way of surgery like in cytoreductive surgery (CRS)+HIPEC. EudraCT: 2018-001035-40.


What is Trained Develops! Theoretical Perspective on Skill Learning.
Knowledge about developmental theories is important for experts or specialists working with children following normal development and children who have various kinds of dysfunction, in order to better understand what happens with processes associated with motor behavior. In this article, we have explored how theories of development and learning can be used to understand processes associated with motor behavior. A probabilistic perspective emphasizes that the changes taking place in the development is a result of interaction: structural changes in the nervous system leading to changes in function and behavior and opposite, functional changes resulting in changes in structure. This bidirectional interaction between biological and experiential aspects is a continuous process which cannot be reduced to either organism or environment. Dynamical systems theory (DST) emphasizes that it is the interaction between the person, the environment, and the task that changes how our movements are, also in terms of how we develop and learn new movements. The interplay between these factors will, over time, lead to changes in motor development. The importance of experience is central to Edelman's theory of neuronal group selection (NGST). Activation of the nervous system increases the connections between certain areas of the brain, and the selection processes in the brain are a result of enhancement of neural connections involved in a "successful" motion. The central nervous system adapts its structure and function in response to internal and external influences, and hence neural plasticity is a prerequisite for learning and development. We argue that Edelman´s approach supports the theory of specificity of learning. From the perspectives of probabilistic epigenesis, DST, and NGST, we can see that being physically active and having the opportunity to get different movement experiences are of great significance for promoting motor development and learning. A variation of purposeful or rewarding physical activity in a variety of contexts will provide individual opportunities for changes of behavior in terms of both quantitative and qualitative changes in motor development.


A Novel 9-Class Auditory ERP Paradigm Driving a Predictive Text Entry System.
Brain-computer interfaces (BCIs) based on event related potentials (ERPs) strive for offering communication pathways which are independent of muscle activity. While most visual ERP-based BCI paradigms require good control of the user's gaze direction, auditory BCI paradigms overcome this restriction. The present work proposes a novel approach using auditory evoked potentials for the example of a multiclass text spelling application. To control the ERP speller, BCI users focus their attention to two-dimensional auditory stimuli that vary in both, pitch (high/medium/low) and direction (left/middle/right) and that are presented via headphones. The resulting nine different control signals are exploited to drive a predictive text entry system. It enables the user to spell a letter by a single nine-class decision plus two additional decisions to confirm a spelled word. This paradigm - called PASS2D - was investigated in an online study with 12 healthy participants. Users spelled with more than 0.8 characters per minute on average (3.4 bits/min) which makes PASS2D a competitive method. It could enrich the toolbox of existing ERP paradigms for BCI end users like people with amyotrophic lateral sclerosis disease in a late stage.


Normal myoblast fusion requires myoferlin.
Muscle growth occurs during embryonic development and continues in adult life as regeneration. During embryonic muscle growth and regeneration in mature muscle, singly nucleated myoblasts fuse to each other to form myotubes. In muscle growth, singly nucleated myoblasts can also fuse to existing large, syncytial myofibers as a mechanism of increasing muscle mass without increasing myofiber number. Myoblast fusion requires the alignment and fusion of two apposed lipid bilayers. The repair of muscle plasma membrane disruptions also relies on the fusion of two apposed lipid bilayers. The protein dysferlin, the product of the Limb Girdle Muscular Dystrophy type 2 locus, has been shown to be necessary for efficient, calcium-sensitive, membrane resealing. We now show that the related protein myoferlin is highly expressed in myoblasts undergoing fusion, and is expressed at the site of myoblasts fusing to myotubes. Like dysferlin, we found that myoferlin binds phospholipids in a calcium-sensitive manner that requires the first C2A domain. We generated mice with a null allele of myoferlin. Myoferlin null myoblasts undergo initial fusion events, but they form large myotubes less efficiently in vitro, consistent with a defect in a later stage of myogenesis. In vivo, myoferlin null mice have smaller muscles than controls do, and myoferlin null muscle lacks large diameter myofibers. Additionally, myoferlin null muscle does not regenerate as well as wild-type muscle does, and instead displays a dystrophic phenotype. These data support a role for myoferlin in the maturation of myotubes and the formation of large myotubes that arise from the fusion of myoblasts to multinucleate myotubes.


Enzymatic and immunological studies of uroporphyrinogen decarboxylase in familial porphyria cutanea tarda and hepatoerythropoietic porphyria.
Uroporphyrinogen decarboxylase activity was measured in hemoglobin-free lysates from two patients with hepatoerythropoietic porphyria (HEP) and from 12 unrelated patients with familial porphyria cutanea tarda (PCT). In HEP patients, enzyme activities were 5% of normal, and familial studies clearly confirmed that patients with HEP are cases of homozygous PCT. Immunoreactive uroporphyrinogen decarboxylase was measured by developing a direct and noncompetitive enzyme immunoassay (EIA). For the 12 familial PCT patients, we found an immunoreactive protein decreased (51%) to the same extent as the catalytic activity (48%) [cross-reactive immunological material ( CRIM ) negative]. The children from the HEP family were also CRIM negative, contrasting with another HEP family previously described as CRIM positive; our data support the hypothesis of a heterogeneity in familial uroporphyrinogen decarboxylase deficiency.


Reversible Photohydration of Trenbolone Acetate Metabolites: Mechanistic Understanding of Product-to-Parent Reversion through Complementary Experimental and Theoretical Approaches.
Photolysis experiments (in H2O and D2O) and quantum chemical calculations were performed to explore the pH-dependent, reversible photohydration of trenbolone acetate (TBA) metabolites. Photohydration of 17α-trenbolone (17α-TBOH) and 17β-trenbolone (17β-TBOH) occurred readily in simulated sunlight to yield hydrated products with incorporated H(+) at C4 and OH(-) at either C5 (5-OH-TBOH) or C12 (12-OH-TBOH) in the tetracyclic steroid backbone. Although unable to be elucidated analytically, theory suggests preferred orientations of cis-12-OH-TBOH (relative to C13 methyl) and trans-5-OH-TBOH, with the former most thermodynamically stable overall. Both experiment and theory indicate limited stability of trans-5-OH-TBOH at acidic pH where it undergoes concurrent, carbocation-mediated thermal rearrangement to cis-12-OH-TBOH and dehydration to regenerate its parent structure. Experiments revealed cis-12-OH-TBOH to be more stable at acidic pH, which is the only condition where its reversion to parent TBA metabolite occurred. At basic pH cis-12-OH-TBOH decayed quickly via hydroxide/water addition, behavior that theory attributes to the formation of a stable enolate resistant to dehydration but prone to thermal hydration. In a noteworthy deviation from predicted theoretical stability, 17α-TBOH photohydration yields major trans-5-OH-TBOH and minor cis-12-OH-TBOH, a distribution also opposite that observed for 17β-TBOH. Because H(+) and OH(-) loss from adjacent carbon centers allows trans-5-OH-TBOH to dehydrate at all pH values, the presumed kinetically controlled yield of 17α-TBOH photohydrates results in a greater propensity for 17α-TBOH reversion than 17β-TBOH. Additional calculations explored minor, but potentially bioactive, trenbolone analogs that could be generated via alternative rearrangement of the acidic carbocation intermediate.


The relative impact of bacterial virulence and host genetic background on cytokine expression during Mycobacterium avium infection of mice.
Resistance to Mycobacterium avium depends on both genetically encoded macrophage functions and acquired T-cell immunity. Cytokines may play a role in either type of resistance. We studied the expression of interleukin-2 (IL-2), IL-4 and interferon-gamma (IFN-gamma) in naturally susceptible BALB/c (Bcgs) and naturally resistant C.D2 (Bcgr) congenic mice infected with two strains of M. avium (one highly virulent and another of low virulence). We observed that cytokine expression patterns correlated better with the virulence of the micro-organism than with the genetic background of the host. The control of the infection by the low virulence strain in either mouse strain was associated with an increased expression of IFN-gamma and IL-2. Only Bcgs mice infected with a virulent strain of M. avium were unable to restrict bacterial growth. An increased expression of IL-4, early during infection, was detected in the course of the latter infection but played no role in determining the susceptibility to infection. Neutralization of IFN-gamma or IL-2 with specific monoclonal antibodies led to an exacerbation of the infection in Bcgr mice by the two strains of M. avium and in Bcgs mice infected with the low virulence strain of M. avium.


Novel multiple sclerosis susceptibility loci implicated in epigenetic regulation.
We conducted a genome-wide association study (GWAS) on multiple sclerosis (MS) susceptibility in German cohorts with 4888 cases and 10,395 controls. In addition to associations within the major histocompatibility complex (MHC) region, 15 non-MHC loci reached genome-wide significance. Four of these loci are novel MS susceptibility loci. They map to the genes L3MBTL3, MAZ, ERG, and SHMT1. The lead variant at SHMT1 was replicated in an independent Sardinian cohort. Products of the genes L3MBTL3, MAZ, and ERG play important roles in immune cell regulation. SHMT1 encodes a serine hydroxymethyltransferase catalyzing the transfer of a carbon unit to the folate cycle. This reaction is required for regulation of methylation homeostasis, which is important for establishment and maintenance of epigenetic signatures. Our GWAS approach in a defined population with limited genetic substructure detected associations not found in larger, more heterogeneous cohorts, thus providing new clues regarding MS pathogenesis.


Measuring willingness-to-pay for risk reduction: an application of conjoint analysis.
This study applies conjoint analysis (CA) to estimate the marginal willingness-to-pay (MWTP) of elderly individuals for a reduction of the risk of fracture of the femur. The good in question is a hypothetical hip protector which lowers the risk of a fracture by different amounts. Other attributes are ease of handling, wearing comfort, and out-of-pocket cost, which are traded off against risk reduction. In 500 face-to-face interviews, pensioners stated whether or not they would buy the product. Results suggest that MWTP for wearing comfort exceeds that for risk reduction. Indeed, willingness-to-pay for the product as a whole is negative, indicating that it should not be included as a mandatory benefit in health insurance.


Phosphate sink containing two-component signaling systems as tunable threshold devices.
Synthetic biology aims to design de novo biological systems and reengineer existing ones. These efforts have mostly focused on transcriptional circuits, with reengineering of signaling circuits hampered by limited understanding of their systems dynamics and experimental challenges. Bacterial two-component signaling systems offer a rich diversity of sensory systems that are built around a core phosphotransfer reaction between histidine kinases and their output response regulator proteins, and thus are a good target for reengineering through synthetic biology. Here, we explore the signal-response relationship arising from a specific motif found in two-component signaling. In this motif, a single histidine kinase (HK) phosphotransfers reversibly to two separate output response regulator (RR) proteins. We show that, under the experimentally observed parameters from bacteria and yeast, this motif not only allows rapid signal termination, whereby one of the RRs acts as a phosphate sink towards the other RR (i.e. the output RR), but also implements a sigmoidal signal-response relationship. We identify two mathematical conditions on system parameters that are necessary for sigmoidal signal-response relationships and define key parameters that control threshold levels and sensitivity of the signal-response curve. We confirm these findings experimentally, by in vitro reconstitution of the one HK-two RR motif found in the Sinorhizobium meliloti chemotaxis pathway and measuring the resulting signal-response curve. We find that the level of sigmoidality in this system can be experimentally controlled by the presence of the sink RR, and also through an auxiliary protein that is shown to bind to the HK (yielding Hill coefficients of above 7). These findings show that the one HK-two RR motif allows bacteria and yeast to implement tunable switch-like signal processing and provides an ideal basis for developing threshold devices for synthetic biology applications.


WE-G-213CD-08: Initial Experience with the Clinical Implementation of a Deep Inspiration Breath Hold for Left Breast Radiotherapy Using Surface Imaging and Visual Aid.
To evaluate the clinical implementation of a deep inspiration breath hold (DIBH) treatment for left breast radiotherapy using surface imaging and visual aid. A CT scan of the patient at DIBH is acquired and used for treatment planning. The plan and skin contour, containing isocenter and surface information are exported from the treatment planning system and imported into the surface imaging system (SIS). The skin contour constitutes the treatment reference surface or target DIBH position. A region of interest (ROI) consisting of the sternum and medial breasts is selected in the SIS. A set of video goggles allows the patient to view their breathing signal within the SIS, aiding in producing a reproducible and stable DIBH similar to simulation. Once the patient is set up at free breathing, she performs a DIBH while being monitored with the SIS. Shifts to minimize displacements from their reference DIBH surface are made. The surface image and patient setup are validated with weekly MV images. The beam is enabled when the two surfaces are within a predetermined tolerance. Data for evaluation of the implementation was acquired for 4 patients throughout treatment. Average treatment time was 16.8 minutes and 14.2 minutes for setup. The average displacement from the reference surface was 0.4 mm during DIBHs. The average reduction of heart mean dose and volume receiving 50% of the prescribed dose between DIBH and FB was 38% and 89% respectively. A total of 15 patients have completed this new treatment. 2 were excluded for inability to achieve reproducible and stable DIBH. The workflow we have implemented has proven to be effective and efficient for clinical purposes. Surface imaging provides adequate real time information valuable to the treatment process. Visual aid has helped patients achieve DIBH with high reproducibility and stability.


Classical and Next-Generation Vaccine Platforms to SARS-CoV-2: Biotechnological Strategies and Genomic Variants.
Several coronaviruses (CoVs) have been identified as human pathogens, including the α-CoVs strains HCoV-229E and HCoV-NL63 and the β-CoVs strains HCoV-HKU1 and HCoV-OC43. SARS-CoV, MERS-CoV, and SARS-CoV-2 are also classified as β-coronavirus. New SARS-CoV-2 spike genomic variants are responsible for human-to-human and interspecies transmissibility, consequences of adaptations of strains from animals to humans. The receptor-binding domain (RBD) of SARS-CoV-2 binds to receptor ACE2 in humans and animal species with high affinity, suggesting there have been adaptive genomic variants. New genomic variants including the incorporation, replacement, or deletion of the amino acids at a variety of positions in the S protein have been documented and are associated with the emergence of new strains adapted to different hosts. Interactions between mutated residues and RBD have been demonstrated by structural modelling of variants including D614G, B.1.1.7, B1.351, P.1, P2; other genomic variants allow escape from antibodies generated by vaccines. Epidemiological and molecular tools are being used for real-time tracking of pathogen evolution and particularly new SARS-CoV-2 variants. COVID-19 vaccines obtained from classical and next-generation vaccine production platforms have entered clinicals trials. Biotechnology strategies of the first generation (attenuated and inactivated virus-CoronaVac, CoVaxin; BBIBP-CorV), second generation (replicating-incompetent vector vaccines-ChAdOx-1; Ad5-nCoV; Sputnik V; JNJ-78436735 vaccine-replicating-competent vector, protein subunits, virus-like particles-NVX-CoV2373 vaccine), and third generation (nucleic-acid vaccines-INO-4800 (DNA); mRNA-1273 and BNT 162b (RNA vaccines) have been used. Additionally, dendritic cells (LV-SMENP-DC) and artificial antigen-presenting (aAPC) cells modified with lentiviral vector have also been developed to inhibit viral activity. Recombinant vaccines against COVID-19 are continuously being applied, and new clinical trials have been tested by interchangeability studies of viral vaccines developed by classical and next-generation platforms.


The skeleton in the closet: should historians of science care about the history of mathematics?
Up until the 1950s, the history of mathematics was an integral part of the history of science. To George Sarton and his contemporaries, mathematics was the rational skeleton that organized science and held it together, and its history was a fundamental component of the broader history of science. But when historians began focusing on the cultural roots of science rather than its rational structure, the study of mathematics was marginalized and ultimately excluded from the history of science. The alienation between the two fields is detrimental to both, and in recent years there has been a sustained effort to reestablish meaningful communication between the two. This time, however, mathematics is seen not as the static skeleton of science but, instead, as a dynamic and historically evolving field in its own right-just like science itself. The new approach allows for a culturally sensitive study of mathematics, as well as a new and fruitful relationship between the history of science and the history of mathematics. The essays in this Focus section offer a sampling of the new approaches, opening the way to a rapprochement between fields that have gone their separate ways but should by rights be closely interconnected.


Ethanol self-administration and alterations in the livers of the cynomolgus monkey, Macaca fascicularis.
Most of the studies of alcoholic liver disease use models in which animals undergo involuntary administration of high amounts of ethanol and consume diets that are often high in polyunsaturated fatty acids. The objectives of this study were (1) to evaluate whether cynomolgus monkeys (Macaca fascicularis) drinking ethanol voluntarily and consuming a diet with moderate amounts of lipid would demonstrate any indices of alcoholic liver disease past the fatty liver stage and (2) to determine whether these alterations were accompanied by oxidative stress. Six adult male and 6 adult female cynomolgus monkeys were allowed to consume ethanol voluntarily for 18 to 19 months. Additional monkeys were maintained on the same consumption protocol, but were not provided with ethanol. During the course of the study, liver biopsy samples were monitored for lipid deposition and inflammation, serum for levels of liver enzymes, and urine for concentrations of the isoprostane (IsoP) metabolite, 2,3-dinor-5,6-dihydro-15-F(2t)-IsoP, a biomarker for oxidative stress. Liver mitochondria were monitored for respiratory control and liver for concentrations of neutral lipids, adenine nucleotides, esterified F(2) isoprostanes, oxidized proteins, 4-hydroxynonenal (HNE)-protein adducts, and protein levels of cytochrome P-450 2E1 and 3A4. Ethanol consumption ranged from 0.9 to 4.05 g/kg/d over the period of the study. Serum levels of aspartate amino transferase were elevated in heavy-consuming animals compared with those in ethanol-naïve or moderate drinkers. Many of the ethanol consumers developed fatty liver and most showed loci of inflammation. Both hepatic energy charge and phosphorylation potential were decreased and NADH-linked respiration was slightly, but significantly depressed in coupled mitochondria as a result of heavy ethanol consumption. The urinary concentrations of 2,3-dinor-5,6-dihydro-15-F(2t)-IsoP increased as high as 33-fold over that observed in ethanol-abstinent animals. Liver cytochrome P-450 2E1 concentrations increased in ethanol consumers, but there were no ethanol-elicited increases in hepatic concentrations of the esterified F(2) isoprostanes, oxidized proteins, or HNE-protein adducts. Our studies show that cynomolgus monkeys undergoing voluntary ethanol consumption for 1.5 years exhibit many of the features observed in the early stages of human alcoholic liver disease. Ethanol-elicited fatty liver, inflammation, and elevated serum aspartate amino transferase were evident with a diet that contained modest amounts of polyunsaturated lipids. The dramatic increases in urinary IsoP demonstrated that the animals were being subjected to significant oxidative stress that correlated with their level of ethanol consumption.


Better Cognition in New Birth Cohorts of 70 Year Olds, But Greater Decline Thereafter.
To evaluate birth cohort differences in level of cognition and rate of change in old age. Data were drawn from three population-based Swedish samples including age-homogenous cohorts born 1901/02, 1906/07, and 1930, and measured on the same cognitive tests at ages 70, 75, and 79 as part of the Gerontological and Geriatric Populations Studies in Gothenburg (H70). We fitted growth curve models to the data using a Bayesian framework and derived estimates and inferences from the marginal posterior distributions. We found moderate to large birth cohort effects in level of performance on all cognitive outcomes. Later born cohorts, however, showed steeper linear rate of decline on reasoning, spatial ability, and perceptual- and motor-speed, but not on picture recognition memory and verbal ability. These findings provide strong evidence for substantial birth cohort effects in cognition in older ages and emphasize the importance of life long environmental factors in shaping cognitive aging trajectories. Inferences from cognitive testing, and standardization of test scores, in elderly populations must take into account the substantial birth cohort differences.


N-Acetylcysteine ethyl ester (NACET): a novel lipophilic cell-permeable cysteine derivative with an unusual pharmacokinetic feature and remarkable antioxidant potential.
Recent large clinical trials failed to confirm the supposed beneficial effects of N-acetylcysteine (NAC) in preventing oxidative stress-related diseases. This may be due to its low bioavailability. We thought that esterification of the carboxyl group of NAC to produce N-acetylcysteine ethyl ester (NACET) would drastically increase the lipophilicity of NAC, thus greatly improving its pharmacokinetics. In the present work, we report on representative chemical, pharmacological and anti-oxidant properties of NACET, especially in direct comparison with its congener NAC. We found that NACET is rapidly absorbed in rats after oral administration but reaches very low concentrations in plasma. This is due to a unique feature of NACET: it rapidly enters the cells where it is trapped being transformed into NAC and cysteine. After oral treatment, NACET (but not NAC) was able to increase significantly the glutathione content of most tissues examined, brain included, and to protect from paracetamol intoxication in the rat. NACET has also the unique feature to accumulate in human erythrocytes where it behaves as a potent protector against hydroperoxide-induced oxidative damage. Our study shows that being able to enter cells and to produce NAC and cysteine, NACET increases circulating hydrogen sulfide (H(2)S), thus representing a good candidate for the oral use as an H(2)S producer, with clear advantages over NAC. NACET has the potential to substitute NAC as a mucolytic agent, as a paracetamol antidote and as a GSH-related antioxidant.


Glioma-derived macrophage migration inhibitory factor (MIF) promotes mast cell recruitment in a STAT5-dependent manner.
Recently, glioma research has increased its focus on the diverse types of cells present in brain tumors. We observed previously that gliomas are associated with a profound accumulation of mast cells (MCs) and here we investigate the underlying mechanism. Gliomas express a plethora of chemoattractants. First, we demonstrated pronounced migration of human MCs toward conditioned medium from cultures of glioma cell lines. Subsequent cytokine array analyses of media from cells, cultured in either serum-containing or -free conditions, revealed a number of candidates which were secreted in high amounts in both cell lines. Among these, we then focused on macrophage migration inhibitory factor (MIF), which has been reported to be pro-inflammatory and -tumorigenic. Infiltration of MCs was attenuated by antibodies that neutralized MIF. Moreover, a positive correlation between the number of MCs and the level of MIF in a large cohort of human glioma tissue samples was observed. Further, both glioma-conditioned media and purified MIF promoted differential phosphorylation of a number of signaling molecules, including signal transducer and activator of transcription 5 (STAT5), in MCs. Inhibition of pSTAT5 signaling significantly attenuated the migration of MCs toward glioma cell-conditioned medium shown to contain MIF. In addition, analysis of tissue microarrays (TMAs) of high-grade gliomas revealed a direct correlation between the level of pSTAT5 in MCs and the level of MIF in the medium. In conclusion, these findings indicate the important influence of signaling cascades involving MIF and STAT5 on the recruitment of MCs to gliomas.


Distinct progenitor origin distinguishes a lineage of dendritic-like cells in spleen.
The dendritic cell (DC) compartment comprises subsets of cells with distinct phenotypes. Previously this lab reported methodology for hematopoiesis of dendritic-like cells in vitro dependent on a murine splenic stromal cell line (5G3). Co-cultures of lineage-depleted bone marrow (Lin(-) BM) over 5G3 continuously produced a distinct population of dendritic-like "L-DC" for up to 35 days. Here the progenitor of L-DC is investigated in relation to known BM-derived hematopoietic progenitors. It is shown here that L-DC-like cells also derive from the CD150(+)Flt3(-) long-term reconstituting-hematopoietic stem cells (HSC), and also from the Flt3(+) multipotential progenitor subset in BM. Lin(-) BM co-cultures also produce a transient population of cells resembling conventional (c) DC. Production of cDC-like cells is shown here to be transient and M-CSF dependent, and also appears following co-culture of described common dendritic progenitors or monocyte dendritic progenitors over 5G3. BM cells from C57BL/6-flt3L(tm1lmx) and C57BL/6-Csf2(tm1Ard) mice which lack cDC precursors and monocytes, are shown here to contain L-DC progenitors which can seed 5G3 co-cultures. L-DC are functionally distinct cells, in that they arise independently of M-CSF, and by direct differentiation from HSC.


Cloning and functional characterization of an NAD(+)-dependent DNA ligase from Staphylococcus aureus.
A Staphylococcus aureus mutant conditionally defective in DNA ligase was identified by isolation of complementing plasmid clones that encode the S. aureus ligA gene. Orthologues of the putative S. aureus NAD(+)-dependent DNA ligase could be identified in the genomes of Bacillus stearothermophilus and other gram-positive bacteria and confirmed the presence of four conserved amino acid motifs, including motif I, KXDG with lysine 112, which is believed to be the proposed site of adenylation. DNA sequence comparison of the ligA genes from wild type and temperature-sensitive S. aureus strain NT64 identified a single base alteration that is predicted to result in the amino acid substitution E46G. The S. aureus ligA gene was cloned and overexpressed in Escherichia coli, and the enzyme was purified to near homogeneity. NAD(+)-dependent DNA ligase activity was demonstrated with the purified enzyme by measuring ligation of (32)P-labeled 30-mer and 29-mer oligonucleotides annealed to a complementary strand of DNA. Limited proteolysis of purified S. aureus DNA ligase by thermolysin produced products with apparent molecular masses of 40, 22, and 21 kDa. The fragments were purified and characterized by N-terminal sequencing and mass analysis. The N-terminal fragment (40 kDa) was found to be fully adenylated. A fragment from residues 1 to 315 was expressed as a His-tagged fusion in E. coli and purified for functional analysis. Following deadenylation with nicotinamide mononucleotide, the purified fragment could self-adenylate but lacked detectable DNA binding activity. The 21- and 22-kDa C-terminal fragments, which lacked the last 76 amino acids of the DNA ligase, had no adenylation activity or DNA binding activity. The intact 30-kDa C terminus of the S. aureus LigA protein expressed in E. coli did demonstrate DNA binding activity. These observations suggest that, as in the case with the NAD(+)-dependent DNA ligase from B. stearothermophilus, two independent functional domains exist in S. aureus DNA ligase, consisting of separate adenylation and DNA binding activities. They also demonstrate a role for the extreme C terminus of the ligase in DNA binding. As there is much evidence to suggest that DNA ligase is essential for bacterial survival, its discovery in the important human pathogen S. aureus indicates its potential as a broad-spectrum antibacterial target for the identification of novel antibiotics.


Acetylcholinesterase (AChE) inhibition aggravates fasting-induced triglyceride accumulation in the mouse liver.
Although fasting induces hepatic triglyceride (TG) accumulation in both rodents and humans, little is known about the underlying mechanism. Because parasympathetic nervous system activity tends to attenuate the secretion of very-low-density-lipoprotein-triglyceride (VLDL-TG) and increase TG stores in the liver, and serum cholinesterase activity is elevated in fatty liver disease, the inhibition of the parasympathetic neurotransmitter acetylcholinesterase (AChE) may have some influence on hepatic lipid metabolism. To assess the influence of AChE inhibition on lipid metabolism, the effect of physostigmine, an AChE inhibitor, on fasting-induced increase in liver TG was investigated in mice. In comparison with ad libitum-fed mice, 30 h fasting increased liver TG accumulation accompanied by a downregulation of sterol regulatory element-binding protein 1 (SREBP-1) and liver-fatty acid binding-protein (L-FABP). Physostigmine promoted the 30 h fasting-induced increase in liver TG levels in a dose-dependent manner, accompanied by a significant fall in plasma insulin levels, without a fall in plasma TG. Furthermore, physostigmine significantly attenuated the fasting-induced decrease of both mRNA and protein levels of SREBP-1 and L-FABP, and increased IRS-2 protein levels in the liver. The muscarinic receptor antagonist atropine blocked these effects of physostigmine on liver TG, serum insulin, and hepatic protein levels of SREBP-1 and L-FABP. These results demonstrate that AChE inhibition facilitated fasting-induced TG accumulation with up regulation of the hepatic L-FABP and SREBP-1 in mice, at least in part via the activation of muscarinic acetylcholine receptors. Our studies highlight the crucial role of parasympathetic regulation in fasting-induced TG accumulation, and may be an important source of information on the mechanism of hepatic disorders of lipid metabolism.


Mapping dissociations in verb morphology.
Substantial behavioural and neuropsychological evidence has been amassed to support the dual-route model of morphological processing, which distinguishes between a rule-based system for regular items (walk-walked, call-called) and an associative system for the irregular items (go-went). Some neural-network models attempt to explain the neuropsychological and brain-mapping dissociations in terms of single-system associative processing. We show that there are problems in the accounts of homogeneous networks in the light of recent brain-mapping evidence of systematic double-dissociation. We also examine the superior capabilities of more internally differentiated connectionist models, which, under certain conditions, display systematic double-dissociations. It appears that the more differentiation models show, the more easily they account for dissociation patterns, yet without implementing symbolic computations.


Immigration and the ephemerality of a natural population bottleneck: evidence from molecular markers.
Population bottlenecks are often invoked to explain low levels of genetic variation in natural populations, yet few studies have documented the direct genetic consequences of known bottlenecks in the wild. Empirical studies of natural population bottlenecks are therefore needed, because key assumptions of theoretical and laboratory studies of bottlenecks may not hold in the wild. Here we present microsatellite data from a severe bottleneck (95% mortality) in an insular population of song sparrows (Melospiza melodia). The major findings of our study are as follows: (i) The bottleneck reduced heterozygosity and allelic diversity nearly to neutral expectations, despite non-random survival of birds with respect to inbreeding and wing length. (ii) All measures of genetic diversity regained pre-bottleneck levels within two to three years of the crash. This rapid recovery was due to low levels of immigration. (iii) The rapid recovery occurred despite a coincident, strong increase in average inbreeding. These results show that immigration at levels that are hard to measure in most field studies can lead to qualitatively very different genetic outcomes from those expected from mutations only. We suggest that future theoretical and empirical work on bottlenecks and metapopulations should address the impact of immigration.


Inhaled nitric oxide and hypoxic respiratory failure in infants with congenital diaphragmatic hernia. The Neonatal Inhaled Nitric Oxide Study Group (NINOS).
We designed and conducted a randomized, double-masked, controlled multicenter study to determine whether inhaled nitric oxide (INO) in term and near-term infants with congenital diaphragmatic hernia (CDH) would reduce the occurrence of death and/or the initiation of extracorporeal membrane oxygenation (ECMO). Infants of 34 weeks gestation or more, <14 days of age with CDH, without known structural heart disease, requiring assisted ventilation for hypoxemic respiratory failure with two oxygenation indices (OIs) of 25 or more at least 15 minutes apart, were eligible for this trial. Infants were centrally randomized and then received masked treatment with 20 ppm NO or 100% oxygen as control. Infants with less than a full response to 20 ppm NO (increase in PaO2 > 20 Torr) after 30 minutes were evaluated at 80 ppm NO/control study gas. The 28 control and 25 treated infants enrolled by the 13 participating centers were not significantly different at randomization for any of the measured variables including prerandomization therapies and initial OIs (45.8 +/- 16.3 for controls, 44.5 +/- 14.5 for INO). Death at <120 days of age or the need for ECMO occurred in 82% of control infants compared with 96% of INO infants (ns). Death occurred in 43% of controls and 48% of the INO group (ns), and ECMO treatment was used for 54% of control and 80% of INO-treated infants. There was no significant improvement in PaO2 (delta PaO2 7.8 +/- 19.8 vs 1.1 +/- 7.6 Torr, ns) nor significant reduction in OI (-2.7 +/- 23.4 vs 4.0 +/- 14.8, ns) associated with INO treatment. Mean peak nitrogen dioxide (NO2) concentration was 1.9 +/- 1.3 ppm and the mean peak methemoglobin was 1.6 +/- 0.8 mg/dL. No infant had study gas discontinued for toxicity. There were no differences between the control and INO groups for the occurrence of intracranial hemorrhage, specific grades of intracranial hemorrhage, periventricular leukomalacia, brain infarction, and pulmonary or gastrointestinal hemorrhages. Although the immediate short-term improvements in oxygenation seen in some treated infants may be of benefit in stabilizing responding infants for transport and initiation of ECMO, we conclude that for term and near-term infants with CDH and hypoxemic respiratory failure unresponsive to conventional therapy, inhaled NO therapy as used in this trial did not reduce the need for ECMO or death.


The Impact of Discriminatory Stress on Changes in Posttraumatic Stress Severity at the Intersection of Race/Ethnicity and Gender.
Given the diversity of military veterans and growing evidence of ethnoracial disparities in posttraumatic stress disorder (PTSD) within this population, elucidating the role of discrimination-related stress in contributing to these disparities is crucial. We examined the relative impact of discriminatory stress (i.e., due to race/ethnicity, religion, nationality, gender, sexual orientation, or physical appearance) on 6-month changes in PTSD symptom severity among trauma-exposed White (74%), Black (11%) and Hispanic/Latino/a/x (15%) veterans (17% female). PTSD symptoms were measured with the 8-item PTSD Checklist for DSM-5. A measure of the extent to which discrimination has caused stress for the respondent assessed discriminatory stress. Hierarchical regression analyses examined interactions among race/ethnicity, gender and discriminatory stress in predicting six-month changes in PTSD severity. Black and Hispanic/Latino/a/x veterans reported higher baseline PTSD severity and discriminatory stress than White veterans, with some variation by gender. Three-way interactions of race/ethnicity by discriminatory stress by gender were significant, controlling for income, education and age. The relationship between discriminatory stress and increases in PTSD severity was significantly stronger for Black women compared with Black men and did not differ between White men and women. There was also a stronger relationship between discriminatory stress and increases in PTSD severity for Hispanic/Latino/x men as compared to Black men. These findings suggest that discriminatory stress impacts PTSD severity differentially for various ethnoracial/gender groups and highlight the value of applying an intersectional framework that accounts for the synergistic connections among multiple identities to future screening, intervention, and research efforts.


Evidence that hsp90 is involved in the altered interactions of Acanthamoeba castellanii variants with bacteria.
There are many similarities between the interactions of environmental protozoa with pathogenic bacterial species and those observed in mammalian macrophages. Since single-celled protozoa predate mammalian hosts, it is likely that interactions in environmental biofilms have selected for many of the bacterial virulence mechanisms responsible for human disease. In order to better understand bacterial-phagocyte interactions, we developed a selection for Acanthamoeba castellanii variants that are more resistant to killing by bacterial pathogens. We identified four amoebal clones that display decreased phagocytosis of bacteria but no difference in uptake of latex beads compared to wild-type amoebae. These amoebal variants display differences in cellular morphology, partial resistance to killing by bacteria, more bactericidal activity, and higher frequencies of lysosome fusion with the bacterial vacuole. Three proteins are present at lower levels in these variants than in wild-type amoebae, and matrix-assisted laser desorption ionization-time of flight mass spectrometry allowed identification of two of them as actin and hsp90. We found that specific inhibitors of hsp90 produce a similar phenotypic effect in macrophages. These data suggest that hsp90 plays a role in phagocytic and, possibly, bactericidal pathways that affect interactions of phagocytic cells with bacteria.


Analysis of ftsZ mutations that confer resistance to the cell division inhibitor SulA (SfiA).
In Escherichia coli, the ftsZ gene is thought to be an essential cell division gene. Several dominant mutations that make lon mutant cells refractory to the cell division inhibitor SulA, sulB9, sulB25, and sfiB114, have been mapped to the ftsZ gene. DNA sequence analysis of these mutations and the sfiB103 mutation confirmed that all of these mutations mapped within the ftsZ gene and revealed that the two sulB mutations were identical and by selection for resistance to higher levels of SulA, contained a second mutation within the ftsZ gene. We therefore propose that these mutations be redesignated ftsZ(Rsa) for resistance to SulA. A procedure involving mutagenesis of ftsZ cloned on low-copy-number vectors was used to isolate three additional ftsZ(Rsa) mutations. DNA sequence analysis of these mutations revealed that they were distinct from the previously isolated mutations. One of these mutations, ftsZ3(Rsa), led to an altered FtsZ protein that could no longer support cell growth but still conferred the Rsa phenotype in the presence of ftsZ+. In addition to being resistant to SulA, all ftsZ(Rsa) mutations also conferred resistance to a LacZ-FtsZ hybrid protein (ZZ). One possibility is that FtsZ functions as a multimer and that FtsZ(Rsa) mutant proteins have an increased ability for multimerization, making them resistant to SulA and ZZ.


Midwives' competence: is it affected by working in a rural location?
Rising health care costs and the need to consolidate expertise in tertiary services have led to the centralisation of services. In the UK, the result has been that many rural maternity units have become midwife-led. A key consideration is that midwives have the skills to competently and confidently provide maternity services in rural areas, which may be geographically isolated and where the midwife may only see a small number of pregnant women each year. Our objective was to compare the views of midwives in rural and urban settings, regarding their competence and confidence with respect to 'competencies' identified as being those which all professionals should have in order to provide effective and safe care for low-risk women. This was a comparative questionnaire survey involving a stratified sample of remote and rural maternity units and an ad hoc comparison group of three urban maternity units in Scotland. Questionnaires were sent to 82 midwives working in remote and rural areas and 107 midwives working in urban hospitals with midwife-led units. The response rate from midwives in rural settings was considerably higher (85%) than from midwives in the urban areas (60%). Although the proportion of midwives who reported that they were competent was broadly similar in the two groups, there were some significant differences regarding specific competencies. Midwives in the rural group were more likely to report competence for breech delivery (p = 0.001), while more urban midwives reported competence in skills such as intravenous fluid replacement (p <0.001) and initial and discharge examination of the newborn (p <0.001). Both groups reported facing barriers to continuing professional development; however, more of the rural group had attended an educational event within the last month (p <0.001). Lack of time was a greater barrier for urban midwives (p = 0.02), whereas distance to training was greater for rural midwives (p = 0.009). Lack of motivation or interest was significantly higher in urban units (p = 0.006). It is often assumed that midwives in rural areas where there are fewer deliveries, will be less competent and confident in their practice. Our exploratory study suggests that the issue of competence is far more complex and deserves further attention.


Transduction of human CD34+ cells that mediate long-term engraftment of NOD/SCID mice by HIV vectors.
Efficient gene transfer into human hematopoietic stem cells (HSCs) is an important goal in the study of the hematopoietic system as well as for gene therapy of hematopoietic disorders. A lentiviral vector based on the human immunodeficiency virus (HIV) was able to transduce human CD34+ cells capable of stable, long-term reconstitution of nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. High-efficiency transduction occurred in the absence of cytokine stimulation and resulted in transgene expression in multiple lineages of human hematopoietic cells for up to 22 weeks after transplantation.


Genome sequence of Aedes aegypti, a major arbovirus vector.
We present a draft sequence of the genome of Aedes aegypti, the primary vector for yellow fever and dengue fever, which at approximately 1376 million base pairs is about 5 times the size of the genome of the malaria vector Anopheles gambiae. Nearly 50% of the Ae. aegypti genome consists of transposable elements. These contribute to a factor of approximately 4 to 6 increase in average gene length and in sizes of intergenic regions relative to An. gambiae and Drosophila melanogaster. Nonetheless, chromosomal synteny is generally maintained among all three insects, although conservation of orthologous gene order is higher (by a factor of approximately 2) between the mosquito species than between either of them and the fruit fly. An increase in genes encoding odorant binding, cytochrome P450, and cuticle domains relative to An. gambiae suggests that members of these protein families underpin some of the biological differences between the two mosquito species.


A Review of Data Quality Assessment in Emergency Medical Services.
Data quality is an important issue in emergency medicine. The unique characteristics of emergency care services, such as high turn-over and the speed of work may increase the possibility of making errors in the related settings. Therefore, regular data quality assessment is necessary to avoid the consequences of low quality data. This study aimed to identify the main dimensions of data quality which had been assessed, the assessment approaches, and generally, the status of data quality in the emergency medical services. The review was conducted in 2016. Related articles were identified by searching databases, including Scopus, Science Direct, PubMed and Web of Science. All of the review and research papers related to data quality assessment in the emergency care services and published between 2000 and 2015 (n=34) were included in the study. The findings showed that the five dimensions of data quality; namely, data completeness, accuracy, consistency, accessibility, and timeliness had been investigated in the field of emergency medical services. Regarding the assessment methods, quantitative research methods were used more than the qualitative or the mixed methods. Overall, the results of these studies showed that data completeness and data accuracy requires more attention to be improved. In the future studies, choosing a clear and a consistent definition of data quality is required. Moreover, the use of qualitative research methods or the mixed methods is suggested, as data users' perspectives can provide a broader picture of the reasons for poor quality data.


Bioabsorbable scaffold for in situ bone regeneration.
A non-porous poly-DL-lactide tubular chamber filled by demineralised bone matrix (DBM) and bone marrow stromal cells (BMSC) in combination, was evaluated as a scaffold for guided bone regeneration (GBR) in an experimental model using the rabbit radius. The tubular chamber had an internal diameter of 4.7 mm, a wall thickness of 0.4 mm and a length of 18 mm. Autologous BMSC were obtained, under general anaesthesia from rabbit iliac crest and isolated by centrifugation technique. Allogenic DBM was obtained from cortico-cancellous bone of rabbits. In general anaesthesia, a 10-mm defect was bilaterally created in the radii of 10 rabbits. On the right side (experimental side) the defect was bridged with the chamber filled with both BMSC and DBM. On the left side (control side) the defect was treated by positioning DBM and BMSC between the two stumps. At an experimental time of 4 months histology and histomorphometry demonstrated that the presence of a tubular chamber significantly improved bone regrowth in the defect The mean thickness of newly-formed bone inside the chamber was about 56.7+/-3.74% of the normal radial cortex, in comparison with 46.7+/-10.7% when DBM and BMSC without the chamber were placed in the defect, P<0.05). These results confirmed the effectiveness of the chamber as a container for factors promoting bone regeneration.


Tetrodotoxin prevents posttraumatic epileptogenesis in rats.
Severe cortical trauma frequently causes epilepsy that develops after a long latency. We hypothesized that plastic changes in excitability during this latent period might be initiated or sustained by the level of neuronal activity in the injured cortex. We therefore studied effects of action potential blockade by application of tetrodotoxin (TTX) to areas of cortical injury in a model of chronic epileptogenesis. Partially isolated islands of sensorimotor cortex were made in 28- to 30-day-old male Sprague-Dawley rats and thin sheets of Elvax polymer containing TTX or control vehicle were implanted over lesions. Ten to 15 days later neocortical slices were obtained through isolates for electrophysiological studies. Slices from all animals (n = 12) with lesions contacted by control-Elvax (58% of 36 slices) exhibited evoked epileptiform field potentials, and those from 4 rats had spontaneous epileptiform events. Only 2 of 11 lesioned animals and 6% of slices from cortex exposed to TTX in vivo exhibited evoked epileptiform potentials, and no spontaneous epileptiform events were observed. There was no evidence of residual TTX during recordings. TTX-Elvax was ineffective in reversing epileptogenesis when implanted 11 days after cortical injury. These data suggest that development of antiepileptogenic drugs for humans may be possible.


Atorvastatin restores endothelial function in offspring of protein-restricted rats in a cholesterol-independent manner.
Maternal protein restriction in rats leads to endothelial dysfunction and decreased NO bioavailability in the offspring. Statins (3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors) are recognized to have pleiotropic actions including increasing NO bioavailability and reducing inflammation and oxidative damage. This study assessed statin treatment on vascular function in a model of endothelial dysfunction, which is independent of dyslipidemia. Wistar rats were fed a control (18% casein) or protein-restricted (9% casein) diet throughout pregnancy. At weaning, a subset of the protein-restricted group was given atorvastatin (10 mg/kg per day) in the drinking water. At 145 days of age, offspring were euthanized by CO(2) inhalation. Plasma samples were collected for markers of inflammation, vascular reactivity of the thoracic aorta, and small mesenteric arteries were assessed on the wire myograph, and tissues were snap frozen for molecular biology analysis. Thoracic aorta endothelial-dependent vasodilatation was attenuated in the male offspring from both protein-restricted groups compared with controls (P<0.05) but was similar in females (P value not significant). Endothelial-dependent dilatation of mesenteric arteries was attenuated in male and female protein-restricted offspring (P<0.05) and was corrected by atorvastatin. Maternal protein restriction increased plasma inflammatory markers granulocyte chemotactic protein, lipocalin-2, and beta(2)-microglobulin in male and C-reactive protein in female offspring (P<0.05). Atorvastatin had no effect on inflammatory markers in the males but restored C-reactive protein to control levels in the females (P<0.05). Aortic and mesenteric artery mRNA levels of endothelial NO synthase, superoxide dismutase 1, and tumor necrosis factor-alpha were unchanged. These data suggest that atorvastatin can restore endothelial function in this model, but its effects are gender specific and dependent on the vascular bed.


Automatic segmentation of the facial nerve and chorda tympani in CT images using spatially dependent feature values.
In cochlear implant surgery, an electrode array is permanently implanted in the cochlea to stimulate the auditory nerve and allow deaf people to hear. A minimally invasive surgical technique has recently been proposed-percutaneous cochlear access-in which a single hole is drilled from the skull surface to the cochlea. For the method to be feasible, a safe and effective drilling trajectory must be determined using a preoperative CT. Segmentation of the structures of the ear would improve trajectory planning safety and efficiency and enable the possibility of automated planning. Two important structures of the ear, the facial nerve and the chorda tympani, are difficult to segment with traditional methods because of their size (diameters as small as 1.0 and 0.3 mm, respectively), the lack of contrast with adjacent structures, and large interpatient variations. A multipart, model-based segmentation algorithm is presented in this article that accomplishes automatic segmentation of the facial nerve and chorda tympani. Segmentation results are presented for ten test ears and are compared to manually segmented surfaces. The results show that the maximum error in structure wall localization is approximately 2 voxels for the facial nerve and the chorda, demonstrating that the method the authors propose is robust and accurate.


Electro-acupuncture promotes survival, differentiation of the bone marrow mesenchymal stem cells as well as functional recovery in the spinal cord-transected rats.
Bone marrow mesenchymal stem cells (MSCs) are one of the potential tools for treatment of the spinal cord injury; however, the survival and differentiation of MSCs in an injured spinal cord still need to be improved. In the present study, we investigated whether Governor Vessel electro-acupuncture (EA) could efficiently promote bone marrow mesenchymal stem cells (MSCs) survival and differentiation, axonal regeneration and finally, functional recovery in the transected spinal cord. The spinal cords of adult Sprague-Dawley (SD) rats were completely transected at T10, five experimental groups were performed: 1. sham operated control (Sham-control); 2. operated control (Op-control); 3. electro-acupuncture treatment (EA); 4. MSCs transplantation (MSCs); and 5. MSCs transplantation combined with electro-acupuncture (MSCs+EA). After 2-8 weeks of MSCs transplantation plus EA treatment, we found that the neurotrophin-3 (NT-3), cAMP level, the differentiation of MSCs, the 5-HT positive and CGRP positive nerve fibers in the lesion site and nearby tissue of injured spinal cord were significantly increased in the MSCs+EA group as compared to the group of the MSCs transplantation or the EA treated alone. Furthermore, behavioral test and spinal cord evoked potentials detection demonstrated a significantly functional recovery in the MSCs +EA group. These results suggest that EA treatment may promote grafted MSCs survival and differentiation; MSCs transplantation combined with EA treatment could promote axonal regeneration and partial locomotor functional recovery in the transected spinal cord in rats and indicate a promising avenue of treatment of spinal cord injury.


Impacts of Satellite Orbit and Clock on Real-Time GPS Point and Relative Positioning.
Satellite orbit and clock corrections are always treated as known quantities in GPS positioning models. Therefore, any error in the satellite orbit and clock products will probably cause significant consequences for GPS positioning, especially for real-time applications. Currently three types of satellite products have been made available for real-time positioning, including the broadcast ephemeris, the International GNSS Service (IGS) predicted ultra-rapid product, and the real-time product. In this study, these three predicted/real-time satellite orbit and clock products are first evaluated with respect to the post-mission IGS final product, which demonstrates cm to m level orbit accuracies and sub-ns to ns level clock accuracies. Impacts of real-time satellite orbit and clock products on GPS point and relative positioning are then investigated using the P3 and GAMIT software packages, respectively. Numerical results show that the real-time satellite clock corrections affect the point positioning more significantly than the orbit corrections. On the contrary, only the real-time orbit corrections impact the relative positioning. Compared with the positioning solution using the IGS final product with the nominal orbit accuracy of ~2.5 cm, the real-time broadcast ephemeris with ~2 m orbit accuracy provided <2 cm relative positioning error for baselines no longer than 216 km. As for the baselines ranging from 574 to 2982 km, the cm-dm level positioning error was identified for the relative positioning solution using the broadcast ephemeris. The real-time product could result in <5 mm relative positioning accuracy for baselines within 2982 km, slightly better than the predicted ultra-rapid product.


Analysis of F1F0-ATPase from Helicobacter pylori.
The adaptive mechanisms that permit Helicobacter species to survive within the gastric mucosa are not well understood. The proton-translocating F1F0-ATPase is an important enzyme for regulating intracellular pH or synthesizing ATP in many other enteric bacteria; therefore, we used degenerate primers derived from conserved bacterial F1F0-ATPase sequences to PCR amplify and clone the gene (atpD) encoding the H. pylori F1F0-ATPase beta subunit. The deduced amino acid sequences of the F1F0-ATPase beta subunits from H. pylori and Wolinella succinogenes were 85% identical (91% similar). To characterize a potential functional role of F1F0-ATPase in H. pylori, H. pylori or Escherichia coli cells were incubated for 60 min in buffered solutions at pH 7, 6, 5, or 4, with or without 100 microM N,N'-dicyclohexylcarbodiimide (DCCD), a specific inhibitor of F1F0-ATPase. At pH 5 and 4, there was no significant decrease in survival of H. pylori in the presence of DCCD compared to its absence, whereas incubation with DCCD at pH 7 and 6 significantly decreased H. pylori survival. E. coli survival was unaffected by DCCD at any pH value tested. We next disrupted the cloned beta-subunit sequence in E. coli by insertion of a kanamycin resistance cassette and sought to construct an isogenic F1F0-ATPase H. pylori mutant by natural transformation and allelic exchange. In multiple transformations of H. pylori cells grown at pH 6 or 7, no kanamycin-resistant F1F0 mutants were isolated, despite consistently successful mutagenesis of other H. pylori genes by using a similar approach and PCR experiments providing evidence for integration of the kanamycin resistance cassette into atpD. The sensitivity of H. pylori to DCCD at pH 7 and 6, and failure to recover F1F0 H. pylori mutants under similar conditions, suggests that the function of this enzyme is required for survival of H. pylori at these pHs.


Anti-Müllerian hormone levels in the spontaneous menstrual cycle do not show substantial fluctuation.
Anti-Müllerian hormone (AMH), a quantitative marker for ovarian reserve, has been suggested to be independent of the classical endocrine fluctuations of the menstrual cycle. The objective of the study was to determine whether AMH levels are constant throughout the menstrual cycle, compared with those of FSH, LH, and estradiol. Frequent blood sampling was performed in 44 fertile, regularly cycling, female volunteers during one full menstrual cycle. The study was conducted at a university hospital. AMH, FSH, LH, and estradiol measurements were allocated to one of seven cycle phases, and a multilevel analysis was performed. Consistent fluctuation patterns were tested by fitting sine patterns to the data. Finally, the frequency in which randomly selected individual samples would remain in one of five preset level categories (quintiles) for each of the variables was studied. A sine pattern fitted to the AMH data was not statistically significant (P = 0.40). In contrast, sine patterns for FSH, LH, and estradiol were highly significant. Comparing the seven cycle phases, no significant differences could be observed between phase-specific AMH levels (P = 0.06). Repeated selection of AMH samples for each individual showed that in 71.5% of selections, AMH values remained in the same quintile, whereas in 27.9% values fell in an adjacent quintile. AMH levels measured through a full menstrual cycle did not show consistent fluctuation patterns in contrast to levels of FSH, LH, and estradiol. Furthermore, random fluctuations were small, indicating that AMH can be relied on as a cycle-independent marker for ovarian reserve.


Globular C1q Receptor (gC1qR/p32/HABP1) Is Overexpressed in Malignant Pleural Mesothelioma and Is Associated With Increased Survival in Surgical Patients Treated With Chemotherapy.
Introduction: Globular C1q receptor (gC1qR/p32/HABP1) is overexpressed in a variety of cancers, particularly adenocarcinomas. This study investigated gC1qR expression in malignant pleural mesothelioma (MPM) and its pathophysiologic correlates in a surgical patient cohort. Methods: Tissue microarrays comprising 6 tumoral and 3 stromal cores from 265 patients with MPM (216 epithelioid, 26 biphasic, and 23 sarcomatoid; 1989-2010) were investigated by immunohistochemistry for gC1qR expression (intensity and distribution by H-score, range 0-300), and immune cell infiltration. Overall survival (OS) was analyzed by the Kaplan-Meier method (high vs. low gC1qR expression delineated by median score) in the whole cohort and by neoadjuvant chemotherapy (NAC) status. Multivariable Cox analysis included stage, chemotherapy, and immune cell infiltration. Results: gC1qR was overexpressed in all histological types of MPMs (263/265, 99.2%) compared to normal pleura. In epithelioid MPM, high gC1qR expression was associated with better OS (median 25 vs. 11 months; p = 0.020) among NAC patients, and among patients without NAC (No-NAC) but who received post-operative chemotherapy (median OS 38 vs. 19 months; p = 0.0007). In multivariable analysis, high gC1qR expression was an independent factor for improved OS in patients treated with NAC. In the No-NAC cohort, high gC1qR expression correlated with lower tumor stage. Moreover, the influence of Ki67 and CD4 T-cell infiltration on OS were more pronounced among patients with high gC1qR expression. Conclusion: This is the first description of gC1qR expression in MPM. The data identify gC1qR as a potential new prognostic factor in patients treated with surgery and chemotherapy.


Psychological responses to continuous terror: a study of two communities in Israel.
The authors evaluated psychological responses to continuous terror. Data were collected after 10 months of escalating hostilities against civilians in Israel. The study's participants were randomly selected adults living in two suburbs of Jerusalem, one frequently and directly exposed to acts of terrorism (N=167) and the other indirectly exposed (N=89). Participants provided information about exposure to terror-related incidents, disruption of daily living, symptoms of posttraumatic stress disorder (PTSD), and general distress (assessed with the Brief Symptom Inventory). Residents of the directly exposed community reported more frequent exposure to terror and deeper disruption of daily living. Notwithstanding, the directly and indirectly exposed groups reported comparable rates of PTSD and similar levels of symptoms: 26.95% of the directly exposed group and 21.35% of the indirectly exposed group met DSM-IV PTSD symptom criteria (criteria B through D), and about one-third of those with PTSD symptoms (35.7% in the directly exposed group and 31.5% in the indirectly exposed group) reported significant distress and dysfunction. Subjects who did not meet PTSD symptom criteria had very low levels of PTSD symptoms, and their Brief Symptom Inventory scores were within population norms. Exposure and disruption of daily living contributed to PTSD symptoms in the directly exposed group. Disruption of daily routines contributed to Brief Symptom Inventory scores in both groups. Continuous terror created similar distress in proximal and remote communities. Exposure to discrete events was not a necessary mediator of terror threat. A subgroup of those exposed developed serious symptoms, whereas others were surprisingly resilient. Disruption of daily routines was a major secondary stressor.


Sciatic inflammatory neuropathy in the rat: surgical procedures, induction of inflammation, and behavioral testing.
Peripheral nerve damage involves inflammation, and is frequently causal to the development of neuropathic pain. However, inflammatory neuropathies often occur in the absence of trauma. We have recently developed an animal model of neuropathic pain where allodynia is induced by nerve inflammation rather than injury. This sciatic inflammatory neuropathy (SIN) model was developed to understand immunologic, neuropathic, and spinal mechanisms underlying allodynia in the territory of the sciatic nerve as well as in extraterritorial and contralateral ("mirror image") sites. A specially designed indwelling catheter system allows immune activators to be selectively injected around one healthy sciatic nerve in awake, behaving rats. Here, we provide detailed procedures on the construction and implantation of chronic indwelling perisciatic catheters used to create SIN. Detailed procedures for implantation of intrathecal catheters via a lumbar vertebra 5 and 6 approach in the same rat are also provided. Methods for testing allodynia and for data analysis are additionally described so to provide all the steps needed for behavioral experimentation.


Patterns of gene duplication in the plant SKP1 gene family in angiosperms: evidence for multiple mechanisms of rapid gene birth.
Gene duplication plays important roles in organismal evolution, because duplicate genes provide raw materials for the evolution of mechanisms controlling physiological and/or morphological novelties. Gene duplication can occur via several mechanisms, including segmental duplication, tandem duplication and retroposition. Although segmental and tandem duplications have been found to be important for the expansion of a number of multigene families, the contribution of retroposition is not clear. Here we show that plant SKP1 genes have evolved by multiple duplication events from a single ancestral copy in the most recent common ancestor (MRCA) of eudicots and monocots, resulting in 19 ASK (Arabidopsis SKP1-like) and 28 OSK (Oryza SKP1-like) genes. The estimated birth rates are more than ten times the average rate of gene duplication, and are even higher than that of other rapidly duplicating plant genes, such as type I MADS box genes, R genes, and genes encoding receptor-like kinases. Further analyses suggest that a relatively large proportion of the duplication events may be explained by tandem duplication, but few, if any, are likely to be due to segmental duplication. In addition, by mapping the gain/loss of a specific intron on gene phylogenies, and by searching for the features that characterize retrogenes/retrosequences, we show that retroposition is an important mechanism for expansion of the plant SKP1 gene family. Specifically, we propose that two and three ancient retroposition events occurred in lineages leading to Arabidopsis and rice, respectively, followed by repeated tandem duplications and chromosome rearrangements. Our study represents a thorough investigation showing that retroposition can play an important role in the evolution of a plant gene family whose members do not encode mobile elements.


Marital status and risk of cardiovascular diseases: a systematic review and meta-analysis.
The influence of marital status on the incidence of cardiovascular disease (CVD) and prognosis after CVD is inconclusive. We systematically reviewed the literature to determine how marital status influences CVD and prognosis after CVD. A search of MEDLINE and Embase in January 2018 without language restriction was performed to identify studies that evaluated the association between marital status and risk of CVD. Search terms related to both marital status and CVD were used and included studies had to be prospective in design. The outcomes of interest were CVD, coronary heart disease (CHD) or stroke incidence and mortality. We performed random effects meta-analysis stratified by the types of population by calculating odds ratios (OR) and 95% confidence intervals (95% CI). Our analysis included 34 studies with more than two million participants. Compared with married participants, being unmarried (never married, divorced or widowed) was associated with increased odds of CVD (OR 1.42; 95% CI 1.00 to 2.01), CHD (OR 1.16,95% CI 1.04 to 1.28), CHD death (OR 1.43,95% CI 1.28 to 1.60) and stroke death (OR 1.55,95% 1.16 to 2.08). Being divorced was associated with increased odds of CHD (P<0.001) for both men and women while widowers were more likely to develop a stroke (P<0.001). Single men and women with myocardial infarction had increased mortality (OR 1.42, 95% CI 1.14 to 1.76) compared with married participants. Marital status appears to influence CVD and prognosis after CVD. These findings may suggest that marital status should be considered in the risk assessment for CVD and outcomes of CVD based on marital status merits further investigation.


Inducing fear using acoustic stimuli-A behavioral experiment on moose (Alces alces) in Sweden.
Prey species may display anti-predatory behavior, i.e., flight, increased vigilance, and decreased feeding, in response to the true presence of a predator or to the implied presence of a predator through, e.g., acoustic cues. In this study, we investigated the anti-predatory reactions of moose (Alces alces) to acoustic stimuli related to hunting, at saltlick stones, a known attractant. In before-during-after-control-impact experiments, we compared the behavioral responses of individuals to: (i) two hunting-related acoustic stimuli-hunting dog barking and human speaking; (ii) nonpredatory acoustic stimuli-bird sounds and; and (iii) no acoustic stimulus (control). We asked: (1) How does the probability of moose leaving the site differ depending on the stimulus they are exposed to?; (2) What affect do the acoustic stimuli have on the amount of time moose spend vigilant, feeding, or away from the site?; and (3) What affect do the stimuli have on the time between events at a site? We found that when exposed to the human stimulus, moose left the sites in 75% of the events, which was significantly more often compared to the dog (39%), bird (24%), or silent (11%) events. If moose did not leave the site, they spent more time vigilant, and less time feeding, particularly when exposed to a dog or human stimulus. Furthermore, moose spent the most time away from the site and took the longest to visit the site again after a human stimulus. Moose were also more likely to leave the site when exposed to the bird stimulus than during silent controls. Those that remained spent more time vigilant, but their behaviors returned to baseline after the bird stimulus ended. These findings suggest that acoustic stimuli can be used to modify the behavior of moose; however, reactions towards presumably threatening and nonthreatening stimuli were not as distinct as we had expected.


262 Voyages Beneath the Sea: a global assessment of macro- and megafaunal biodiversity and research effort at deep-sea hydrothermal vents.
For over 40 years, hydrothermal vents and the communities that thrive on them have been a source of profound discovery for deep-sea ecologists. These ecosystems are found throughout the world on active plate margins as well as other geologically active features. In addition to their ecologic interest, hydrothermal vent fields are comprised of metallic ores, sparking a nascent industry that aims to mine these metal-rich deposits for their mineral wealth. Here, we provide the first systematic assessment of macrofaunal and megafaunal biodiversity at hydrothermal vents normalized against research effort. Cruise reports from scientific expeditions as well as other literature were used to characterize the extent of exploration, determine the relative biodiversity of different biogeographic provinces, identify knowledge gaps related to the distribution of research effort, and prioritize targets for additional sampling to establish biodiversity baselines ahead of potential commercial exploitation. The Northwest Pacific, Southwest Pacific, and Southern Ocean biogeographic provinces were identified as high biodiversity using rarefaction of family-level incidence data, whereas the North East Pacific Rise, Northern East Pacific, Mid-Atlantic Ridge, and Indian Ocean provinces had medium biodiversity, and the Mid-Cayman Spreading Center was identified as a province of relatively low biodiversity. A North/South divide in the extent of biological research and the targets of hydrothermal vent mining prospects was also identified. Finally, we provide an estimate of sampling completeness for each province to inform scientific and stewardship priorities.


Screening of biomarkers of drug resistance or virulence in ESCAPE pathogens by MALDI-TOF mass spectrometry.
Rapid identification and characterisation of drug-resistant bacterial pathogens have an important role in diagnostic and antimicrobial stewardship. Response time in the diagnosis of not only the etiological agent but also in antimicrobial susceptibility results is of utmost importance in patient treatment. In this study, matrix-assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectrometry (MS) was used to screen for biomarkers of ESCAPE (vancomycin-resistant Enterococcus faecium, methicillin-resistant Staphylococcus aureus, hypervirulent NAP1/ribotype 027 Clostridioides [Clostridium] difficile, multidrug resistant Acinetobacter baumannii, multidrug resistant Pseudomonas aeruginosa, and carbapenem-resistant Enterobacteriaceae) pathogens to predict antimicrobial resistance or hypervirulence. Several biomarkers of drug-resistant genotypes in S. aureus, A. baumannii, P. aeruginosa, and K. pneumoniae, as well as hypervirulence in C. difficile, were detected. The fastest possible susceptibility testing with MALDI-TOF MS is simultaneous detection of a characteristic drug-resistant peak and species identification in the same spectra generated in routine processing. According to our approach, resistance or virulence biomarker peaks can be identified while performing routine microbiology analysis, and no additional assays nor prolonged incubation time is needed. Outstanding biomarker peaks detected in our study should be further analysed by additional methods to identify the specific proteins involved.


Trunk movements in human locomotion.
Trunk movements in the frontal and sagittal planes were studied in 10 healthy males (18-35 yrs) during normal walking (1.0-2.5 m/s) and running (2.0-6.0 m/s) on a treadmill. Movements were recorded with a Selspot optoelectronic system. Directions, amplitudes and phase relationships to the stride cycle (defined by the leg movements) were analyzed for both linear and angular displacements. During one stride cycle the trunk displayed two oscillations in the vertical (mean net amplitude 2.5-9.5 cm) and horizontal, forward-backward directions (mean net amplitude 0.5-3 cm) and one oscillation in the lateral, side to side direction (mean net amplitude 2-6 cm). The magnitude and timing of the various oscillations varied in a different way with speed and mode of progression. Differences in amplitudes and timing of the movements at separate levels along the spine gave rise to angular oscillations with a similar periodicity as the linear displacements in both planes studied. The net angular trunk tilting in the frontal plane increased with speed from 3-10 degrees. The net forward-backward trunk inclination showed a small increase with speed up to 5 degrees in fast running. The mean forward inclination of the trunk increased from 6 degrees to about 13 degrees with speed. Peak inclination to one side occurred during the support phase of the leg on the same side. Peak forward inclination was reached at the initiation of the support phase in walking, whereas in running the peak inclination was in the opposite direction at this point. The adaptations of trunk movements to speed and mode of progression could be related to changing mechanical conditions and different demands on equilibrium control due to e.g. changes in support phase duration and leg movements.


Low muscle mass in older men: the role of lifestyle, diet and cardiovascular risk factors.
To explore associations between low muscle mass and a wide range of lifestyle, dietary and cardiovascular risk factors in older men including metabolic risk factors, markers of inflammation, endothelial dysfunction and coagulation. Cross-sectional study. British Regional Heart Study. 4252 men aged 60-79 years. PARTICIPANTS attended a physical examination in 1998-2000, and completed a general questionnaire and a food frequency questionnaire. Low muscle mass was assessed by two measures: midarm muscle circumference (MAMC) and fat-free mass index (FFMI). Associations between risk factors and low muscle mass were analysed using logistic regression. Physical inactivity, insulin resistance, C-reactive protein, von Willebrand factor and fibrinogen were associated with significantly increased odds of low MAMC and FFMI after adjustment for body mass index, lifestyle characteristics and morbidity. Those with higher percent energy intake from carbohydrates showed decreased odds of low MAMC (OR: 0.73, 95% CI: 0.55-0.96) and FFMI (OR: 0.76, 95% CI: 0.58-0.99). Other dietary variables, smoking, alcohol intake, D-dimer, interleukin 6 and homocysteine showed no important associations with MAMC and FFMI. Increasing physical activity, consuming a diet with a high proportion of energy from carbohydrates, and taking steps to prevent insulin resistance and reduce inflammation and endothelial dysfunction may help to reduce the risk of low muscle mass in older men.


Relationship between the logistic EuroSCORE and the Society of Thoracic Surgeons Predicted Risk of Mortality score in patients implanted with the CoreValve ReValving system--a Bern-Rotterdam Study.
Surgical risk scores, such as the logistic EuroSCORE (LES) and Society of Thoracic Surgeons Predicted Risk of Mortality (STS) score, are commonly used to identify high-risk or "inoperable" patients for transcatheter aortic valve implantation (TAVI). In Europe, the LES plays an important role in selecting patients for implantation with the Medtronic CoreValve System. What is less clear, however, is the role of the STS score of these patients and the relationship between the LES and STS. The purpose of this study is to examine the correlation between LES and STS scores and their performance characteristics in high-risk surgical patients implanted with the Medtronic CoreValve System. All consecutive patients (n = 168) in whom a CoreValve bioprosthesis was implanted between November 2005 and June 2009 at 2 centers (Bern University Hospital, Bern, Switzerland, and Erasmus Medical Center, Rotterdam, The Netherlands) were included for analysis. Patient demographics were recorded in a prospective database. Logistic EuroSCORE and STS scores were calculated on a prospective and retrospective basis, respectively. Observed mortality was 11.1%. The mean LES was 3 times higher than the mean STS score (LES 20.2% +/- 13.9% vs STS 6.7% +/- 5.8%). Based on the various LES and STS cutoff values used in previous and ongoing TAVI trials, 53% of patients had an LES > or =15%, 16% had an STS > or =10%, and 40% had an LES > or =20% or STS > or =10%. Pearson correlation coefficient revealed a reasonable (moderate) linear relationship between the LES and STS scores, r = 0.58, P < .001. Although the STS score outperformed the LES, both models had suboptimal discriminatory power (c-statistic, 0.49 for LES and 0.69 for STS) and calibration. Clinical judgment and the Heart Team concept should play a key role in selecting patients for TAVI, whereas currently available surgical risk score algorithms should be used to guide clinical decision making.


Evolution of resistance to a last-resort antibiotic in Staphylococcus aureus via bacterial competition.
Antibiotic resistance is a key medical concern, with antibiotic use likely being an important cause. However, here we describe an alternative route to clinically relevant antibiotic resistance that occurs solely due to competitive interactions among bacterial cells. We consistently observe that isolates of Methicillin-resistant Staphylococcus aureus diversify spontaneously into two distinct, sequentially arising strains. The first evolved strain outgrows the parent strain via secretion of surfactants and a toxic bacteriocin. The second is resistant to the bacteriocin. Importantly, this second strain is also resistant to intermediate levels of vancomycin. This so-called VISA (vancomycin-intermediate S. aureus) phenotype is seen in many hard-to-treat clinical isolates. This strain diversification also occurs during in vivo infection in a mouse model, which is consistent with the fact that both coevolved phenotypes resemble strains commonly found in clinic. Our study shows how competition between coevolving bacterial strains can generate antibiotic resistance and recapitulate key clinical phenotypes.


Age-related small vessel disease: a potential contributor to neurodegeneration in multiple sclerosis.
Multiple sclerosis (MS) is a chronic inflammatory demyelinating disorder of the central nervous system wherein, after an initial phase of transient neurological defects, slow neurological deterioration due to progressive neuronal loss ensues. Age is a major determinant of MS progression onset and disability. Over the past years, several mechanisms have been proposed to explain the key drivers of neurodegeneration and disability accumulation in MS. However, the effect of commonly encountered age-related cerebral vessel disease, namely small vessel disease (SVD), has been largely neglected and constitutes the aim of this review. SVD shares some features with MS, that is, white matter demyelination and brain atrophy, and has been shown to contribute to the neuronal damage seen in vascular cognitive impairment. Several lines of evidence suggest that an interaction between MS and SVD may influence MS-related neurodegeneration. SVD may contribute to hypoperfusion, reduced vascular reactivity and tissue hypoxia, features seen in MS. Venule and endothelium abnormalities have been documented in MS but the role of arterioles and of other neurovascular unit structures, such as the pericyte, has not been explored. Vascular risk factors (VRF) have recently been associated with faster progression in MS, though the mechanisms are unclear since very few studies have addressed the impact of VRF and SVD on MS imaging and pathology outcomes. Therapeutic agents targeting the microvasculature and the neurovascular unit may impact both SVD and MS and may benefit patients with dual pathology.


Tetrazepam: a benzodiazepine which dissociates sedation from other benzodiazepine activities. I. Psychopharmacological profile in rodents.
In the 1,4-benzodiazepine (BZD) series the nature of the C(5) substituent is critical for activity. In tetrazepam this substituent is a cyclohexenyl ring, in all other clinically effective 1,4-BZD derivatives it is a phenyl ring. The activities of tetrazepam and diazepam, whose chemical structures differ only by the nature of the C(5) substituent, were compared in rodent models which are predictive of anticonvulsant, anxiolytic, muscle relaxant and sedative effects. In mice, tetrazepam and diazepam antagonized pentylenetetrazol (PTZ)-induced seizures, increased novel food consumption, decreased rearing behavior in the Staircase test and impaired performance in the Traction test. The effects of both drugs were abolished by Ro 15-1788. In rats, both drugs antagonized PTZ-induced seizures, released punished responding in an approach-avoidance conflict procedure and blocked the PTZ discriminative cue. In mice and rats, and with both BZDs, maximal activity occurred 15 to 30 min after drug administration. In mice and rats, the overall anxiolytic and muscle relaxant potencies of tetrazepam were one-fourth and one-sixth those of diazepam, respectively. The anticonvulsant potency of tetrazepam varied from one-half (rats) to one-twentieth (mice) that of diazepam. In rats and mice, the sedative potency of tetrazepam (Rotorod test; locomotor activity) was approximately one-sixteenth that of diazepam. Finally, tetrazepam induced a loss of righting reflex in mice, with an ED50 value which was approximately 70-times greater than that of diazepam. It is concluded that replacing the 5-phenyl ring by a 5-cyclohexenyl ring leads to a relative dissociation of the pharmacological actions of the 1,4-BZDs.


Alkaloids from the root of Indonesian Annona muricata L.
Annona muricata L. has been used traditionally in Indonesia to treat disease. Phytochemical studies on the alkaloid fractions from the root of Annona muricata L. from Malang-Indonesia resulted in the isolation of an unreported benzylisoquinoline alkaloid (+)-xylopine 5 as well as four known alkaloids (1-4). The crude methanol extract and alkaloid fractions were tested against Plasmodium falciparum K1 and against bacteria (Escherichia coli, Klebsiella pneumonia, Acinetobacter buamanii, Pseudomonas aeruginosa, Methicillin-resistant Staphylococcus aureus) with insignificant activities (MIC > 32 µg/mL). Individual alkaloids were tested against a human suspension cancer cell line (HL-60 leukemia cells) and two human fibroblastic cancer cell lines (A549 lung cancer cells and HepG2 liver cancer cells) in which compound 5 was the most toxic alkaloid with IC50 values ranging from 20 to 80 µM.


Validation of the Brazilian version of the 2007 Youth Risk Behavior Survey.
To validate the psychometric properties of the Brazilian version of the 2007 Youth Risk Behavior Survey (YRBS) questionnaire. The original version of the 2007 YRBS was translated into Portuguese and back-translated into English. The questionnaire versions were analyzed by a committee of experts. The committee used semantic, idiomatic, cultural and conceptual equivalences as criteria of analysis. The final version of the translated 2007 YRBS questionnaire was administered in two occasions, with an interval of two weeks, in a sample of 873 high school students of both sexes, in the city of Londrina, Southern Brazil, to identify the psychometric properties. Test-retest reliability was analyzed by calculating the Kappa index of agreement and prevalence rate of each type of risk behavior when application was repeated. Chi-square test was used to identify statistical differences between the first and second questionnaire applications. After minor changes identified in the translation process, the committee of experts concluded that the Portuguese version of the 2007 YRBS showed semantic, idiomatic, cultural and conceptual equivalences. Significant differences between the prevalence rates of both applications were found in 23.4% of items. The identification of 91% of items with moderate-to-substantial Kappa index of agreement and mean value of this index of 68.6% indicated the quality of psychometric properties of the Portuguese version of the 2007 YRBS. The translation, cross-cultural adaptation and psychometric qualities of the 2007 YRBS questionnaire were satisfactory, thus enabling its application in epidemiological studies in Brazil.


Characterization of Pdgfrb-Cre transgenic mice reveals reduction of ROSA26 reporter activity in remodeling arteries.
With the intention to modulate gene expression in vascular mural cells of remodeling vessels, we generated and characterized transgenic mouse lines with Cre recombinase under the control of the platelet-derived growth factor receptor-β promoter, referred to as Tg(Pdgfrb-Cre)(35Vli) . Transgenic mice were crossed with the Gt(ROSA)26Sor(tm1Sor) strain and examined for Cre activation by β-galactosidase activity, which was compared with endogenous Pdgfrb expression. In addition, Pdgfrb-Cre mice were used to drive expression of a conditional myc-tagged Cthrc1 transgene. There was good overlap of β-galactosidase activity with endogenous Pdgfrb immunoreactivity. However, dedifferentiation of vascular mural cells induced by carotid artery ligation revealed a dramatic discrepancy between ROSA26 reporter activity and Pdgfrb promoter driven Cre dependent myc-tagged Cthrc1 transgene expression. Our studies demonstrate the capability of the Pdgfrb-Cre mouse to drive conditional transgene expression as a result of prior Cre-mediated recombination in tissues known to express endogenous Pdgfrb. In addition, the study shows that ROSA26 promoter driven reporter mice are not suitable for lineage marking of smooth muscle in remodeling blood vessels.