Between-task competition for intentions and actions.
People can switch quickly and flexibly from one task to another, but suffer the effects of between-task competition when they do so: After switching, they tend to be distracted by irrelevant stimulus information and hampered by incorrect actions associated with recently performed tasks. This competition results in performance costs of switching, as well as a bias against switching when there is choice over which task to perform, particularly when switching from a difficult task to an easier one. Two experiments investigated the locus of these between-task competition effects in voluntary task switching. Participants switched between an easy location classification and a harder shape classification, making two responses on each trial: the first to register their task choice, the second to perform the chosen task on a subsequently presented stimulus. The results indicated that participants chose to perform the difficult shape task more often than the easier location task, evidence that between-task competition affects intentions that are expressed independently of task-specific actions. The bias was stronger in participants with faster choice speed, suggesting that these influences are relatively automatic. Moreover, even though participants had unlimited time to choose and prepare a task before stimulus presentation, their subsequent performance was nonetheless sensitive to persisting effects of between-task competition. Altogether these results indicate the pervasive influence of between-task competition, which affects both the expression of global task intentions and the production of task-specific actions.


Serotonin neurons derived from rhesus monkey embryonic stem cells: similarities to CNS serotonin neurons.
We sought an in vitro primate model for serotonin neurons. Rhesus monkey embryonic stem (ES) cell colonies were isolated and differentiated into embryoid bodies (EBs), then transferred to serum-free medium with 1% insulin-transferrin-selenium for 7 days to induce neural precursor cell (NPC) formation. NPCs were cultured in medium with 1% N-2 neural supplement and human fibroblast growth factor 2 (FGF2, 10 ng/ml) for 7 days to stimulate cell proliferation. Lastly, NPCs were dispersed into single cells and cultured without FGF2 for another 7 days to obtain terminal differentiation. Terminal cells were characterized for neuronal and serotonergic markers. Over 95% of the NPCs were immunopositive for nestin and Musashi1. Terminally differentiated cells appeared in both small and large morphologies. Most (>95%) of the mature cells (both small and large) were immunopositive for neuron-specific nuclear protein (NeuN), synaptophysin, microtubule-associated protein (MAP2C), Tau-1, neurofilament 160 (NF-160), beta-tubulin (TujIII), tryptophan hydroxylase (TPH), serotonin, the serotonin reuptake transporter (SERT), estrogen receptor-beta (ERbeta), and progestin receptor (PR), but not estrogen receptor-alpha (ERalpha). Less than 2-3% of cells were positive for tyrosine hydroxylase (TH). Reverse transcriptase polymerase chain reaction (RT-PCR) detected mRNA transcripts for TPH-1, TPH-2, SERT, 5-HT1A-autoreceptor, ERbeta, and PR in the differentiated population. A low level of expression of ERalpha mRNA was also detected. Quantitative RT-PCR indicated that the relative abundance of TPH-2 mRNA was greater than TPH-1 mRNA. Serotonin as measured by ELISA increased 3-fold in the mature stage compared to the selection and expansion stages. In summary, a remarkably high percentage of cells derived from monkey ES cells exhibited neuronal plus serotonergic markers as well as nuclear steroid receptors similar to primate CNS serotonin neurons, suggesting that these cells may serve as a useful primate model for serotonergic neurons.


Sexual Dimorphism in Stress-induced Hyperthermia in SNAP25Δ3 mice, a mouse model with disabled Gβγ regulation of the exocytotic fusion apparatus.
Behavioral assays in the mouse can show marked differences between male and female animals of a given genotype. These differences identified in such preclinical studies may have important clinical implications. We recently made a mouse model with impaired presynaptic inhibition through Gβγ-SNARE signaling. Here, we examine the role of sexual dimorphism in the severity of the phenotypes of this model, the SNAP25Δ3 mouse. In males, we already reported that SNAP25Δ3 homozygotes demonstrated phenotypes in motor coordination, nociception, spatial memory and stress processing. We now report that while minimal sexually dimorphic effects were observed for the nociceptive, motor or memory phenotypes, large differences were observed in the stress-induced hyperthermia paradigm, with male SNAP25Δ3 homozygotes exhibiting an increase in body temperature subsequent to handling relative to wild-type littermates, while no such genotype-dependent effect was observed in females. This suggests sexually dimorphic mechanisms of Gβγ-SNARE signaling for stress processing or thermoregulation within the mouse. Second, we examined the effects of heterozygosity with respect to the SNAP25Δ3 mutation. Heterozygote SNAP25Δ3 animals were tested alongside homozygote and wild-type littermates in all of the aforementioned paradigms and displayed phenotypes similar to wild-type animals or an intermediate state. From this, we conclude that the SNAP25Δ3 mutation does not behave in an autosomal dominant manner, but rather displays incomplete dominance for many phenotypes.


Public financing of health in developing countries: a cross-national systematic analysis.
Government spending on health from domestic sources is an important indicator of a government's commitment to the health of its people, and is essential for the sustainability of health programmes. We aimed to systematically analyse all data sources available for government spending on health in developing countries; describe trends in public financing of health; and test the extent to which they were related to changes in gross domestic product (GDP), government size, HIV prevalence, debt relief, and development assistance for health (DAH) to governmental and non-governmental sectors. We did a systematic analysis of all data sources available for government expenditures on health as agent (GHE-A) in developing countries, including government reports and databases from WHO and the International Monetary Fund (IMF). GHE-A consists of domestically and externally financed public health expenditures. We assessed the quality of these sources and used multiple imputation to generate a complete sequence of GHE-A. With these data and those for DAH to governments, we estimated government spending on health from domestic sources. We used panel-regression methods to estimate the association between government domestic spending on health and GDP, government size, HIV prevalence, debt relief, and DAH disbursed to governmental and non-governmental sectors. We tested the robustness of our conclusions using various models and subsets of countries. In all developing countries, public financing of health in constant US$ from domestic sources increased by nearly 100% (IMF 120%; WHO 88%) from 1995 to 2006. Overall, this increase was the product of rising GDP, slight decreases in the share of GDP spent by government, and increases in the share of government spending on health. At the country level, while shares of government expenditures to health increased in many regions, they decreased in many sub-Saharan African countries. The statistical analysis showed that DAH to government had a negative and significant effect on domestic government spending on health such that for every US$1 of DAH to government, government health expenditures from domestic resources were reduced by $0.43 (p=0) to $1.14 (p=0). However, DAH to the non-governmental sector had a positive and significant effect on domestic government health spending. Both results were robust to multiple specifications and subset analyses. Other factors, such as debt relief, had no detectable effect on domestic government health spending. To address the negative effect of DAH on domestic government health spending, we recommend strong standardised monitoring of government health expenditures and government spending in other health-related sectors; establishment of collaborative targets to maintain or increase the share of government expenditures going to health; investment in the capacity of developing countries to effectively receive and use DAH; careful assessment of the risks and benefits of expanded DAH to non-governmental sectors; and investigation of the use of global price subsidies or product transfers as mechanisms for DAH. Bill & Melinda Gates Foundation.


Salinity induction of recycling Crassulacean acid metabolism and salt tolerance in plants of Talinum triangulare.
Crassulacean acid metabolism (CAM) can be induced by salinity, thus conferring the plant higher water-use efficiency. Talinum triangulare does not frequently encounter salt in its natural habitat but is cultivated in soils that may become salinized. Here we examined whether plants of T. triangulare can grow in saline soils and show salt-induced CAM. Leaf gas exchange, carbon isotopic ratio (δ13C), nocturnal acid accumulation (ΔH+), water relations, photosynthetic pigment and mineral contents, leaf anatomy and growth were determined in greenhouse in plants irrigated with 0, 150, 300 and 400 mm NaCl. Salinity reduced gas exchange and induced CAM, ΔH+ reaching 50.2 μmol H+ g-1 fresh mass under 300 mm NaCl. No nocturnal CO2 uptake, but compensation, was observed. Values of δ13C were lowest under 0 and 400 mm NaCl, and highest under 150 and 300 mm. The difference in osmotic potential (ψs) between control and treated plants averaged 0.45 MPa for the three [NaCl] values, the decrease in ψs being accounted for by up to 63 % by Na+ and K+. Pigment contents were unaffected by treatment, suggesting lack of damage to the photosynthetic machinery. Changes in stomatal index with unchanged stomatal density in newly expanded leaves suggested inhibited differentiation of epidermal cells into stomata. Whole-leaf and parenchymata thickness increased under 150 and 300 mm NaCl. Only plants irrigated with 400 mm NaCl showed reductions in biomass (stems, 41 %; reproductive structures, 78 %). The K/Na molar ratio decreased with [NaCl] from 2.0 to 0.4. The operation of CAM in the recycling mode was evidenced by increased ΔH+ with no nocturnal CO2 uptake. Talinum triangulare can be classified as a halo-tolerant species based on its low K/Na molar ratio under salinity and the relatively small reduction in growth only at the highest [NaCl].


Dynamic protein adsorption in microchannels by "stop-flow" and continuous flow.
This work addresses two ways of loading proteins on microchannel surfaces for immunoassay applications: the "stop-flow" and the continuous flow processes. The "stop-flow" method consists of successive static incubation periods where the bulk solution depletes upon the adsorption process. In the present paper, a multi-step "stop-flow" protein coating is studied and compared to a coating under continuous flow conditions. For the "stop-flow", a non-dimensional parameter is here introduced, indicating the adsorbing capacity of the system, by which it is possible to calculate the number of loads necessary to reach the optimum coverage. For the continuous flow, the effects on the adsorption of the kinetic rates, flow velocity and wall capacity have been considered. This study shows the importance of a careful choice of the fluid velocity to minimise the sample waste. For diffusion controlled and kinetics controlled processes, two flow velocity criteria are provided in order to obtain the best possible coverage, with the same amount of sample as with the "stop-flow".


Bibliotherapy for depression.
Bibliotherapy can be used to treat mild to moderate depression or subthreshold depressive symptoms, as a sole or supplementary therapy. Bibliotherapy is a form of guided self-help. The patient works through a structured book, independently from the doctor. The role of the doctor is to support and motivate the patient as they continue through the book and to help clarify any questions or concerns the patient may have. Relevant books can be purchased or often borrowed from a library, with limited cost and good accessibility from a patient perspective. Patients need to have a reading age above 12 years and have a positive attitude toward self-help. Bibliotherapy has NHMRC Level 1 evidence of efficacy and no serious adverse effects have been reported. This article forms part of a series on non-drug treatments, which summarise the indications, considerations and the evidence, and where clinicians and patients can find further information.


Variation of the enhanced biologically damaging solar UV due to clouds.
The variation of the biologically damaging solar UV (UVBE) enhanced by clouds above that of clear sky UVBE has been investigated. This was undertaken for summer through to winter for SZA of 5 to 60 degrees employing an integrated automatic cloud and spectral UV measurement system that recorded the solar UV spectra and the sky images at five minute intervals. The UVBE calculated with action spectra with higher relative effectiveness in the UVA produced the lower percentage of cloud enhanced cases. The DNA UVBE provided the highest percentage of cloud enhanced cases compared to the total number of UV scans with 2.2% cloud enhanced cases. As a comparison, the plant and fish melanoma UVBE provided the lowest percentage of cloud enhanced cases with 0.6 to 0.8% cloud enhanced cases. For the cases of cloud enhanced UVBE, the average ratio of the measured UVBE to calculated cloud free UVBE for the photokeratitis, cataracts, plant, generalized plant damage and fish melanoma action spectra was 1.21 to 1.25. In comparison, the highest value of 1.4 was for the DNA action spectrum.


Shifts in ophthalmic care utilization during the COVID-19 pandemic in the US.
Healthcare restrictions during the COVID-19 pandemic, particularly in ophthalmology, led to a differential underutilization of care. An analytic approach is needed to characterize pandemic health services usage across many conditions. A common analytical framework identified pandemic care utilization patterns across 261 ophthalmic diagnoses. Using a United States eye care registry, predictions of utilization expected without the pandemic were established for each diagnosis via models trained on pre-pandemic data. Pandemic effects on utilization were estimated by calculating deviations between observed and expected patient volumes from January 2020 to December 2021, with two sub-periods of focus: the hiatus (March-May 2020) and post-hiatus (June 2020-December 2021). Deviation patterns were analyzed using cluster analyses, data visualizations, and hypothesis testing. Records from 44.62 million patients and 2455 practices show lasting reductions in ophthalmic care utilization, including visits for leading causes of visual impairment (age-related macular degeneration, diabetic retinopathy, cataract, glaucoma). Mean deviations among all diagnoses are 67% below expectation during the hiatus peak, and 13% post-hiatus. Less severe conditions experience greater utilization reductions, with heterogeneities across diagnosis categories and pandemic phases. Intense post-hiatus reductions occur among non-vision-threatening conditions or asymptomatic precursors of vision-threatening diseases. Many conditions with above-average post-hiatus utilization pose a risk for irreversible morbidity, such as emergent pediatric, retinal, or uveitic diseases. We derive high-resolution insights on pandemic care utilization in the US from high-dimensional data using an analytical framework that can be applied to study healthcare disruptions in other settings and inform efforts to pinpoint unmet clinical needs.


Identifying which septic patients have increased mortality risk using severity scores: a cohort study.
Early aggressive therapy can reduce the mortality associated with severe sepsis but this relies on prompt recognition, which is hindered by variation among published severity criteria. Our aim was to test the performance of different severity scores in predicting mortality among a cohort of hospital inpatients with sepsis. We anonymously linked routine outcome data to a cohort of prospectively identified adult hospital inpatients with sepsis, and used logistic regression to identify associations between mortality and demographic variables, clinical factors including blood culture results, and six sets of severity criteria. We calculated performance characteristics, including area under receiver operating characteristic curves (AUROC), of each set of severity criteria in predicting mortality. Overall mortality was 19.4% (124/640) at 30 days after sepsis onset. In adjusted analysis, older age (odds ratio 5.79 (95% CI 2.87-11.70) for ≥80y versus <60y), having been admitted as an emergency (OR 3.91 (1.31-11.70) versus electively), and longer inpatient stay prior to sepsis onset (OR 2.90 (1.41-5.94) for >21d versus <4d), were associated with increased 30 day mortality. Being in a surgical or orthopaedic, versus medical, ward was associated with lower mortality (OR 0.47 (0.27-0.81) and 0.26 (0.11-0.63), respectively). Blood culture results (positive vs. negative) were not significantly association with mortality. All severity scores predicted mortality but performance varied. The CURB65 community-acquired pneumonia severity score had the best performance characteristics (sensitivity 81%, specificity 52%, positive predictive value 29%, negative predictive value 92%, for 30 day mortality), including having the largest AUROC curve (0.72, 95% CI 0.67-0.77). The CURB65 pneumonia severity score outperformed five other severity scores in predicting risk of death among a cohort of hospital inpatients with sepsis. The utility of the CURB65 score for risk-stratifying patients with sepsis in clinical practice will depend on replicating these findings in a validation cohort including patients with sepsis on admission to hospital.


Inverse diffraction grating of Maxwell's equations in biperiodic structures.
Consider a time-harmonic electromagnetic plane wave incident on a perfectly conducting biperiodic surface (crossed grating). The diffraction is modeled as a boundary value problem for the three-dimensional Maxwell equation. The surface is assumed to be a small and smooth deformation of a planar surface. In this paper, a novel approach is developed to solve the inverse diffraction grating problem in the near-field regime, which is to reconstruct the surface with resolution beyond Rayleigh's criterion. The method requires only a single incident field with one polarization, one frequency, and one incident direction, and is realized by using the fast Fourier transform. Numerical results show that the method is simple, efficient, and stable to reconstruct biperiodic surfaces with subwavelength resolution.


New technologies for glycomic analysis: toward a systematic understanding of the glycome.
Carbohydrates are the most difficult class of biological molecules to study by high-throughput methods owing to the chemical similarities between the constituent monosaccharide building blocks, template-less biosynthesis, and the lack of clearly identifiable consensus sequences for the glycan modification of cohorts of glycoproteins. These molecules are crucial for a wide variety of cellular processes ranging from cell-cell communication to immunity, and they are altered in disease states such as cancer and inflammation. Thus, there has been a dedicated effort to develop glycan analysis into a high-throughput analytical field termed glycomics. Herein we highlight major advances in applying separation, mass spectrometry, and microarray methods to the fields of glycomics and glycoproteomics. These new analytical techniques are rapidly advancing our understanding of the importance of glycosylation in biology and disease.


Successful Transabdominal Removal of Penetrating Iron Rod in the Rectum: A Case Report.
Foreign bodies in the anus and rectum are not uncommon presentations globally. Reasons for foreign bodies in the rectum can be trauma, assault, psychiatric reasons but the most common reason documented is sexual pleasure, and objects range from sex toys to tools to packed drugs. Regardless of the reason, health care providers must maintain nonjudgmental composure and express empathy. Numerous cases have been reported of anorectal foreign body due to various causes. Removal of the objects has mostly been through rectally but some does need surgical intervention. A multidisciplinary approach and radiologic investigations are important to guide in the management outline. Establishment of guidelines for anorectal foreign bodies are needed to guide surgeons and emergency physicians on the course of treatment. We present a case of an eleven-year old school boy slid and fell on an iron rod that penetrated his rectum through his anal canal. Presented with clinical features of peritonitis, where emergency laparotomy was done and the iron rod was extracted abdominally with primary repair of the rectum. The boy recovered well and was discharged four days after with no complications.


Sample of a client intake information protocol: a synopsis and rationale.
The utilization of standardized comprehensive forms in the field of orofacial myology is crucial as this profession continues to grow and establish assessment and treatment protocols. This article formally presents a comprehensive health history intake form currently in use, and highlights the rationale for each particular question within this form in an effort to explore the evidence-based theory behind each question utilized. Highlighting the importance of obtaining a thorough health history as it pertains to our profession, personally allows the clinician to ultimately best plan a therapeutic strategy and assess the individual criteria necessary for successful orofacial myofunctional habituation.


Survival of infants ≤24 months of age with brain tumors: A population-based study using the SEER database.
Brain tumors are the most common solid malignancy and leading cause of cancer-related deaths in infants. Current epidemiological data is limited by low numbers of reported cases. This study used a population-based approach with analysis of contemporary and historical survival curves to provide up-to-date prognostication. Observational cohort analysis was performed using the Surveillance, Epidemiology and End Results (SEER) database. Infants with brain tumors diagnosed from 1973 to 2013 were categorized by the most common tumor types (diffuse astrocytic and oligodendroglioma, choroid plexus, embryonal, ependymal, medulloblastoma and pilocytic astrocytoma). The 1, 5 and 10 year survival was stratified by decade, with trends in management and outcomes analyzed. We identified 2996 affected infants satisfying inclusion criteria. All tumor types, except embryonal and choroid plexus, demonstrated improving survival with time. Infants with embryonal tumors showed a decline in survival from the 1970s to 1990s (p = 0.009), whereas infants with choroid plexus tumors had no change in survival. Infants with ependymal tumors experienced the greatest improvement in survival from 1980s to 1990s and 1990s to 2000s (p = 0.0001, p = 0.01), with 5-year survival probability improving from 28% (95% CI 15-42%) in the 1980s to 77% (95% CI 69-83%) the 2000s. The use of radiation declined from 1970 to 2000 for all tumors; however, radiation treatment for embryonal and ependymal subtypes increased after 2000. While overall survival for infants with brain tumors has improved from the 1970s onwards, not every tumor type has seen a statistically significant change. Given changes in management and survival, prognostication of infants with brain tumor should be updated.


Looking beyond changes in averages in evaluating foundational learning: Some inequality measures.
This paper uses measurements of learning inequality to explore whether learning interventions that are aimed at improving means also reduce inequality, and if so, under what conditions. There is abundant evidence that learning levels are generally low in low- and middle-income countries (LMIC), but there is less knowledge about how learning achievement is distributed within these contexts, and especially about how these distributions change as mean levels increase. We use child-level data on foundational literacy outcomes to quantitatively explore whether and how learning inequality using metrics borrowed from the economics and inequality literature can help us understand the impact of learning interventions. The paper deepens recent work in several ways. First, it extends the analysis to six LMIC, displaying which measures are computable and coherent across contexts and baseline levels. This extension can add valuable information to program evaluation, without being redundant with other metrics. Second, we show the large extent to which the disaggregation of inequality of foundational skills between- and within-schools and grades varies by context and language. Third, we present initial empirical evidence that, at least in the contexts of analysis of foundational interventions, improving average performance can reduce inequality as well, across all levels of socioeconomic status (SES). The data show that at baseline, the groups with the highest internal inequality tend to be the groups with lowest SES and lowest reading scores, as inequality among the poor themselves is higher than among their wealthier counterparts. Regardless of which SES groups benefit more in terms of a change in mean levels of reading, there is still a considerable reduction in inequality by baseline achievement as means increase. These results have policy implications in terms of targeting of interventions: much can be achieved in terms of simultaneously improving averages and increasing equality. This seems particularly true when the initial learning levels are as low as they currently are the developing world.


Broadband terahertz wave generation from a MgO:LiNbO3 ridge waveguide pumped by a 1.5 μm femtosecond fiber laser.
Cherenkov phase-matched terahertz (THz) wave generation from a MgO:LiNbO3 ridge waveguide was studied using optical rectification. Pumping was achieved using 20 and 60 fs laser pulses from a fiber laser centered at 1.56 μm. Time-domain spectroscopy (TDS) results showed a single-cycle pulse with 20 fs pulse pumping and a near-single-cycle pulse with 60 fs pulse pumping. The spectrum covered the range of 0.1-7 THz, with a signal-to-noise ratio of over 50 dB. The output power measured by a Si bolometer and a deuterated triglycine sulfate pyroelectric detector is shown and compared to that of a commercial photoconductive antenna. This system is believed to be a promising THz source for low-cost, compact, robust, and highly integrated TDS, THz imaging, and tomography systems.


Gastric HER2 Testing Study (GaTHER): an evaluation of gastric/gastroesophageal junction cancer testing accuracy in Australia.
Trastuzumab provides a survival benefit in patients with human epidermal growth factor receptor 2 (HER2)-amplified/overexpressed advanced gastric and gastroesophageal junction cancers (GC/GJCs). However, the optimal method for testing is unclear. The aim of this study was to assess interlaboratory agreement on HER2 scoring in GC/GJC to aid the development of a robust testing algorithm for diagnostic practice in Australia. Nine laboratories assessed the HER2 status of 100 GC/GJC tissue samples by immunohistochemistry (IHC) and in situ hybridization (ISH) [chromogenic (CISH) or silver (SISH)] using both HER2 copy number and HER2:chr17 (chromosome 17) ratio. Results were compared with reference fluorescence ISH (FISH). Interlaboratory agreement on IHC3+ scoring was good (κ=0.76), and there was good/very good agreement between IHC (positivity defined as IHC3+) and ISH when HER2 copy number was used (κ=0.72 to 0.87). Agreement on CISH/SISH scoring was good/very good when HER2 copy number was used (κ=0.68 to 0.86), and agreement between CISH/SISH and FISH using HER2 copy number was very good (κ=0.88 to 0.91). Agreement was reduced when HER2:chr17 ratio was used. The good agreement for HER2 copy number determined by bright-field ISH suggests that this is the optimal method for testing in GC/GJC cases. An IHC3+ score was strongly predictive of a positive ISH result, although agreement for all IHC scores was only moderate, suggesting that IHC triage before ISH testing would be the most cost-effective strategy. However, because of the unique features of GC/GJC samples and the difficulty of ensuring consistent HER2 staining in the community setting, it is recommended that HER2 status in advanced GC/GJC be determined by both IHC and ISH in the same laboratory.


Atomic view of a toxic amyloid small oligomer.
Amyloid diseases, including Alzheimer's, Parkinson's, and the prion conditions, are each associated with a particular protein in fibrillar form. These amyloid fibrils were long suspected to be the disease agents, but evidence suggests that smaller, often transient and polymorphic oligomers are the toxic entities. Here, we identify a segment of the amyloid-forming protein αB crystallin, which forms an oligomeric complex exhibiting properties of other amyloid oligomers: β-sheet-rich structure, cytotoxicity, and recognition by an oligomer-specific antibody. The x-ray-derived atomic structure of the oligomer reveals a cylindrical barrel, formed from six antiparallel protein strands, that we term a cylindrin. The cylindrin structure is compatible with a sequence segment from the β-amyloid protein of Alzheimer's disease. Cylindrins offer models for the hitherto elusive structures of amyloid oligomers.


M. tuberculosis Rv2252 encodes a diacylglycerol kinase involved in the biosynthesis of phosphatidylinositol mannosides (PIMs).
Phosphorylated lipids play important roles in biological systems, not only as structural moieties but also as modulators of cellular function. Phospholipids of pathogenic bacteria are known to play roles both as membrane components and as factors that modulate the infectious process. Mycobacterium tuberculosis is, however, noteworthy in that it has an extremely diverse repertoire of biologically active phosphorylated lipids that, in the absence of a specialized protein translocation system, appear to constitute the main means of communication with the host. Many of these lipids are derived from phosphatidylinositol (PI) that is differentially processed to give rise to phosphatidylinositol mannosides (PIMs) or lipoarabinomannan. In preliminary studies on the lipid processing enzymes associated with the bacterial cell wall, a kinase activity was noted that gave rise to a novel lipid species released by the bacterium. It was determined that this kinase activity was encoded by the ORF Rv2252. Rv2252 demonstrates the capacity to phosphorylate various amphipathic lipids of host and bacterial origin, in particular a M. tuberculosis derived diacylglycerol. Targeted deletion of the rv2252 gene resulted in disruption of the production of certain higher order PIM species, suggesting a role for Rv2252 in the biosynthetic pathway of PI, a PIM precursor.


Atmospheric concentrations of polycyclic aromatic hydrocarbons during chimney sweeping.
Air sampled from the breathing zone of chimney sweeps during "dirty work" and soot samples were analysed for polycyclic aromatic hydrocarbons (PAH). A total of 20 PAH were quantified by gas chromatography-mass spectrometry in 115 air samples and 18 soot samples. These included benzo(b)fluoranthene, benzo(a)pyrene (BaP), chrysene, dibenz(a,h)anthracene, and indeno (1,2,3-cd)pyrene, all of which are animal carcinogens. The summed atmospheric concentration of these compounds depended on the type of fuel used and averaged 2.27 micrograms/m3 for oil fuel. If a mixture of oil and solid fuel was used the concentration was 5.06 micrograms/m3; pure solid fuel heating yielded 5.08 micrograms/m3. The air concentrations of BaP were 0.36, 0.83, and 0.82 micrograms/m3 respectively. The soot samples recovered after using the three different fuel types were 10.50, 109.10, and 51.25 mg BaP/kg. The maximum total concentrations of the five carcinogenic PAH were 243.70, 691.06, and 213.94 mg/kg respectively. The time weighted, shift mean concentrations of 0.02 to 0.21 micrograms/m3 benzo(a)pyrene obtained on 11 days form the basis for the industrial medical estimation of risk.


Known glioma risk loci are associated with glioma with a family history of brain tumours -- a case-control gene association study.
Familial cancer can be used to leverage genetic association studies. Recent genome-wide association studies have reported independent associations between seven single nucleotide polymorphisms (SNPs) and risk of glioma. The aim of this study was to investigate whether glioma cases with a positive family history of brain tumours, defined as having at least one first- or second-degree relative with a history of brain tumour, are associated with known glioma risk loci. One thousand four hundred and thirty-one glioma cases and 2,868 cancer-free controls were identified from four case-control studies and two prospective cohorts from USA, Sweden and Denmark and genotyped for seven SNPs previously reported to be associated with glioma risk in case-control designed studies. Odds ratios were calculated by unconditional logistic regression. In analyses including glioma cases with a family history of brain tumours (n = 104) and control subjects free of glioma at baseline, three of seven SNPs were associated with glioma risk: rs2736100 (5p15.33, TERT), rs4977756 (9p21.3, CDKN2A-CDKN2B) and rs6010620 (20q13.33, RTEL1). After Bonferroni correction for multiple comparisons, only one marker was statistically significantly associated with glioma risk, rs6010620 (ORtrend for the minor (A) allele, 0.39; 95% CI: 0.25-0.61; Bonferroni adjusted ptrend , 1.7 × 10(-4) ). In conclusion, as previously shown for glioma regardless of family history of brain tumours, rs6010620 (RTEL1) was associated with an increased risk of glioma when restricting to cases with family history of brain tumours. These findings require confirmation in further studies with a larger number of glioma cases with a family history of brain tumours.


The effect of Tmem135 overexpression on the mouse heart.
Tissues with high-energy demand including the heart are rich in the energy-producing organelles, mitochondria, and sensitive to mitochondrial dysfunction. While alterations in mitochondrial function are increasingly recognized in cardiovascular diseases, the molecular mechanisms through which changes in mitochondria lead to heart abnormalities have not been fully elucidated. Here, we report that transgenic mice overexpressing a novel regulator of mitochondrial dynamics, transmembrane protein 135 (Tmem135), exhibit increased fragmentation of mitochondria and disease phenotypes in the heart including collagen accumulation and hypertrophy. The gene expression analysis showed that genes associated with ER stress and unfolded protein response, and especially the pathway involving activating transcription factor 4, are upregulated in the heart of Tmem135 transgenic mice. It also showed that gene expression changes in the heart of Tmem135 transgenic mice significantly overlap with those of aged mice in addition to the similarity in cardiac phenotypes, suggesting that changes in mitochondrial dynamics may be involved in the development of heart abnormalities associated with aging. Our study revealed the pathological consequence of overexpression of Tmem135, and suggested downstream molecular changes that may underlie those disease pathologies.


Endothelial function in HIV-infected patients receiving protease inhibitor therapy: does immune competence affect cardiovascular risk?
The use of HIV protease inhibitors (PIs) as a component of combination antiretroviral therapy in HIV-infected patients has been associated with dyslipidaemia, but its significance as a risk factor for cardiovascular disease is unclear. Endothelial dysfunction is an early phase of atherogenesis that may be assessed non-invasively with ultrasonography in vivo. To evaluate vascular function and investigate potential determinants of endothelial dysfunction of the peripheral circulation in PI-treated, HIV-infected men with dyslipidaemia. Observational, case-control study. We studied 24 HIV-infected, PI-treated men with dyslipidaemia and 24 normolipidaemic, healthy male controls matched for age and body mass index. Brachial artery endothelial function was studied using high-resolution ultrasound and computerized edge-detection software. This non-invasive technique measured post-ischaemic flow-mediated dilatation (FMD), and the endothelium-independent vasodilatory response to glyceryl trinitrate (GTN). Within the HIV patient group, FMD was significantly associated with percentage of 'naïve' CD4 + 45RA + T cells (p = 0.03), while plasma lipid/lipoprotein and insulin levels, body mass, and smoking status did not correlate with endothelial function. FMD was not significantly different between the study group and the controls. The atherogenic potential of PI-associated dyslipidaemia may be attenuated in HIV-infected patients with decreased immune competence, reflecting a possible contribution of cell-mediated immune responses to the pathogenesis of atherosclerosis.


On the role of micro-porosity in affecting the environmental stability of atomic/molecular layer deposited (ZnO)a(Zn-O-C6H4-O)b films.
Atomic/molecular layer deposited (ALD/MLD) inorganic-organic thin films form a novel class of materials with tunable properties. In selected cases, hybrid materials are reported to show environmental instability, specifically towards moisture. In this article, we focus on zinc oxide/zincone multi-layers with the theoretical formula of (ZnO)a(Zn-O-C6H4-O)b. We show by means of ellipsometric porosimetry that micro-porosity in the range of 0.42 and 2 nm in the pristine zincone layer is responsible for its environmental degradation. During degradation, it is found that a relative micro-porosity content of 1.2 ± 0.1 vol% in the pristine zincone films evolves into micro-mesoporosity with a relative content of 39 ± 1 vol%. We also show that the micro-porosity in the zincone layer can be gradually suppressed when few cycles (a = 1-10) of ZnO are introduced. The resulting (ZnO)a(Zn-O-C6H4-O)b = 1 periodic multilayer is an environmentally stable film with a = 10. It is found that the suppressed micro-porosity is due to the development of continuous ZnO layers with a≥ 10.


Effect of Continuous Dewaxed Brown Rice Ingestion on the Cognitive Function of Elderly Individuals.
Dewaxed brown rice (DBR), which is prepared by removing only the outermost wax layer of brown rice using a new rice milling technique, has improved water absorbency, digestibility, and taste compared to regular brown rice. DBR has a nutritional value close to that of brown rice, including a rich amount of lipopolysaccharides that are known to improve cognitive function in mice. This study thus aimed to verify the influence of continuous DBR ingestion on cognitive function among elderly individuals. The present study employed a crossover comparison design using the Revised Hasegawa Dementia Scale to assess cognitive function. Our findings confirmed that long-term DBR ingestion contributed to the prevention and reduction of overall cognitive decline, especially among elderly individuals with low cognitive function. Thus, DBR has the potential to be a useful staple food that maintains brain homeostasis among elderly individuals.


Overweight and obesity in twenty-year-old Swedes in relation to birthweight and weight development during childhood.
To describe the frequency of overweight and obesity from birth to 20 years of age and analyse weight at 20 years of age in relation to weight and weight development during early childhood and adolescence. A longitudinal, population-based study, which followed 496 children from birth to 20 years of age. Information about weight and height was collected from health records at child health centres and school health care. At 20 years of age, weight and height measurements were taken by one of the authors. At 20 years of age, 124 (25%) of the youth were obese or overweight. Of these youths, 60% had normal weight at 5.5 years. Of the teenagers who were overweight/obese at 15 years, 79% remained overweight/obese at 20 years of age. Out of the 124 overweight/obese at 20, 47% had normal weight at 15 years. [Corrections added after online publication on April 18, 2012: 'Out of the 124 obese at 20' has been changed to 'Out of the 124 overweight/obese at 20']. No relation was found between rapid weight gain during preschool age and overweight and obesity in 20-year-olds. The majority of those who were overweight/obese at 20 years of age were recruited after 5.5 years of age, and half of them in their late teens. Thus, during the preschool period, the entire population should be the target of primary prevention from overweight/obesity and, in the case of teenagers, prevention strategies should be developed for the whole population as well as treatment strategies for teenagers with established overweight/obesity.


The nature and management of labor pain: part II. Pharmacologic pain relief.
A group of family physicians, obstetricians, midwives, obstetric anesthesiologists, and childbirth educators attended an evidence-based symposium in 2001 on the nature and management of labor pain and discussed a series of systematic reviews that focused on methods of labor pain management. Parenteral opioids provide modest pain relief in labor, and little evidence supports the use of one agent over another. Epidural analgesia is used during labor in most large U.S. hospitals, and its use is rapidly increasing in small hospitals. Although epidural analgesia is the most effective form of pain relief, its use is associated with a longer labor, an increased incidence of maternal fever, and increased rates of operative vaginal delivery. The effect of epidural analgesia on rates of cesarean delivery is controversial. Nitrous oxide provides a modest analgesic effect, but it is used less often in the United States than in other developed nations. Paracervical block provides effective analgesia in the first stage of labor, but its use is limited by postblock bradycardia. Research is needed regarding which pain-relief options women would choose if they were offered a range of choices beyond epidural analgesia or parenteral opioids.


Impact of proton pump inhibitors on efficacy of antiplatelet strategies with ticagrelor or aspirin after percutaneous coronary intervention: Insights from the GLOBAL LEADERS trial.
Several studies have suggested that proton pump inhibitors (PPIs) may reduce the antiplatelet effects of clopidogrel and/or aspirin, possibly leading to cardiovascular events. We aimed to investigate the association between PPI and clinical outcomes in patients treated with ticagrelor monotherapy or conventional antiplatelet therapy after percutaneous coronary intervention (PCI). This is a subanalysis of the randomized GLOBAL LEADERS trial, comparing the experimental antiplatelet arm (23-month ticagrelor monotherapy following 1-month dual antiplatelet therapy [DAPT]) with the reference arm (12-month aspirin monotherapy following 12-month DAPT) after PCI. Patient-oriented composite endpoints (POCEs: all-cause mortality, myocardial infarction, stroke, or repeat revascularization) and its components were assessed stratified by PPI use as a time-dependent covariate in patients with the experiment or reference antiplatelet arm. Among 15,839 patients, 2115 patients (13.5%) experienced POCE at 2 years. In the reference arm, the use of PPIs was independently associated with POCE (hazard ratio [HR]: 1.27; 95% confidence interval [CI]: 1.12-1.44) and its individual components, whereas it was not in the experimental arm (HR: 1.04; 95% CI: 0.92-1.19; pinteraction = 0.035). During the second-year follow-up, patients taking aspirin with PPIs had a significantly higher risk of POCE compared to those on aspirin without PPIs (HR: 1.57; 95% CI: 1.27-1.94), whereas the risk did not differ significantly irrespective of PPI in ticagrelor monotherapy group (HR: 1.03; 95% CI: 0.83-1.28; pinteraction = 0.008). In contrast to conventional antiplatelet strategy, there were no evidence suggesting the interaction between ticagrelor monotherapy and PPIs on increased cardiovascular events, which should be confirmed in further studies. URL: https://clinicaltrials.gov.


Heterogeneity of photosynthetic membranes from Rhodobacter capsulatus: size dispersion and ATP synthase distribution.
The density distribution of photosynthetic membrane vesicles (chromatophores) from Rhodobacter capsulatus has been studied by isopicnic centrifugation. The average vesicle diameters, examined by electron microscopy, varied between 61 and 72 nm in different density fractions (70 nm in unfractionated chromatophores). The ATP synthase catalytic activities showed maxima displaced toward the higher density fractions relative to bacteriochlorophyll, resulting in higher specific activities in those fractions (about threefold). The amount of ATP synthase, measured by quantitative Western blotting, paralleled the catalytic activities. The average number of ATP synthases per chromatophore, evaluated on the basis of the Western blotting data and of vesicle density analysis, ranged between 8 and 13 (10 in unfractionated chromatophores). Poisson distribution analysis indicated that the probability of chromatophores devoid of ATP synthase was negligible. The effects of ATP synthase inhibition by efrapeptin on the time course of the transmembrane electric potential (evaluated as carotenoid electrochromic response) and on ATP synthesis were studied comparatively. The ATP produced after a flash and the total charge associated with the proton flow coupled to ATP synthesis were more resistant to efrapeptin than the initial value of the phosphorylating currents, indicating that several ATP synthases are fed by protons from the same vesicle.


Pharmacokinetics of butixocort 21-propionate, budesonide, and beclomethasone dipropionate in the rat after intratracheal, intravenous, and oral treatments.
The disposition of butixocort 21-propionate (JO 1222) in the rat after intratracheal treatment with a pulmonary aerosol was compared to that of budesonide (BUD) and beclomethasone dipropionate (BDP) using tritium-labeled aerosols. Each aerosol canister delivered 50 micrograms of 3H-labeled steroid through an intratracheal catheter directly into the lung. Rats were sacrificed at different times after treatment. Lung and plasma concentrations of unchanged drug and active metabolites were measured by HPLC using a radioactivity detector. Tritium-labeled drugs (1.5 mg/kg) also were administered by the iv and oral routes for the purpose of comparing their systemic availability. The extent of biotransformation of all three steroids in the lung was very limited. JO 1222 was metabolized to the active compounds butixocort (JO 1717) and butixocort 21-methyl (JO 1605). BDP was transformed primarily to beclomethasone monopropionate (BMP); BUD was not metabolized in the lung. In contrast to these results, large differences were observed between plasma concentrations and systemic availability of the three drugs. BUD was rapidly absorbed from the lung, and its plasma elimination half-life was about 3 hr. BDP was hydrolyzed rapidly into its active metabolite BMP after intratracheal and iv treatments; BDP was not observed in plasma after oral treatment. Additionally, plasma concentrations of BMP were higher than those of BUD administered at the same doses. Assuming that BDP was transformed entirely into BMP, the oral bioavailability of BMP at 1.5 mg/kg was around 72%, while that of BUD was 15%. JO 1222 has a distinct pharmacokinetic profile due to the extensive metabolic clearance of both the unchanged drug and its active metabolites JO 1717 and JO 1605.(ABSTRACT TRUNCATED AT 250 WORDS)


Is There an Association Between Indication for Intrauterine Balloon Tamponade and Balloon Failure?
Objective Determine whether the indication for intrauterine balloon tamponade (IUBT) is associated with failure rates. Study Design Cohort study of women who underwent IUBT for postpartum hemorrhage (PPH) from 2007 to 2014. The indication was categorized as uterine atony or placental-site bleeding. Primary outcome was IUBT failure, defined as the need for uterine artery embolization or hysterectomy. Secondary outcomes were estimated blood loss (EBL) after balloon placement, transfusion of red blood cells (RBC), transfusion of fresh frozen plasma (FFP) and/or cryoprecipitate, and intensive care unit (ICU) admission. Results 306 women underwent IUBT: 241 (78.8%) for uterine atony and 65 (21.2%) for placental site bleeding. Overall, 67 (21.9%) women experienced IUBT failure. The frequency of failure was similar in those with uterine atony compared with those with placental-site bleeding (21.2 vs 24.6%, p = 0.55). This finding persisted after adjusting for potential confounders (aOR, 0.97; 95% CI, 0.48-1.99). Median EBL after balloon placement (190 [interquartile range, 93-375] vs 195 [interquartile range, 103-500] mL, p = 0.46), and frequencies of RBC transfusion (62.7 vs 66.2%, p = 0.60), FFP and/or cryoprecipitate transfusion (25.3 vs 33.8%, p = 0.17), and ICU admission (12.4 vs 16.9%, p = 0.35) were also similar. Conclusion IUBT was similarly effective for managing PPH from uterine atony or placental-site bleeding.


Resolving the heading-direction ambiguity in vertical-beam radar observations of migrating insects.
Each year, massive numbers of insects fly across the continents at heights of hundreds of meters, carried by the wind, bringing both environmental benefits and serious economic and social costs. To investigate the insects' flight behavior and their response to winds, entomological radar has proved to be a particularly valuable tool; however, its observations of insect orientation are ambiguous with regard to the head/tail direction, and this greatly hinders interpretation of the migrants' flight behavior.We have developed two related methods of using wind data to resolve the head/tail ambiguity, and we have compared their outputs with those from simply assigning the heading direction to be that which is closer to the track direction. We applied all three methods to observations of Australian plague locust migrations made with an insect monitoring radar.For the study dataset, some of the headings selected by the simpler method are shown to be clearly incorrect. The two new methods generally agree and reveal a significantly different, and presumably more accurate, relationship of heading direction to track direction. However, use of these methods leads to quite a large proportion of the sample being lost because the wind values, which derive from a regional-scale numerical model, are shown to be incompatible with the radar observations. This exploratory study has moreover demonstrated that locusts are frequently oriented at a large angle to their track and that quite often their movement is at least slightly tailfirst.Both new methods appear to be a significant improvement on the simpler method. As well as providing an accurate representation of migratory flight behavior, they allow occasions when the model wind values are unreliable to be eliminated from the data sample.


Tacrolimus as a liver flush solution to ameliorate the effects of ischemia/reperfusion injury following liver transplantation.
The goal of this report is to evaluate in a prospective randomized fashion the effect of flushing hepatic allografts with tacrolimus before transplantation. A prospective, double-blinded, randomized trial was performed. Twenty patients receiving orthotopic liver transplants from October 2000 to October 2001 were randomized into two groups. Group 1 (active) was administered tacrolimus, 20 ng/mL, plus Plasma-lyte A (Baxter Healthcare Corp, Deerfield, IL) liver flush solution; and group 2 (placebo) was administered only Plasma-lyte A. Ischemia/reperfusion injury was assessed in both groups after transplantation by means of serum laboratory values to assess hepatocellular damage, synthetic function, and ion transport capacity. Peak values were recorded for each parameter, and their distributions were compared. There were no statistically significant differences between groups for age, sex, total ischemia time, or cause of liver disease. Global multivariate comparison of peak changes in all measures of liver function indicated liver injury was significantly lower with tacrolimus treatment than placebo (P =.01). The sample median for group 1 was less than for group 2 in all parameters measured. Individual statistical comparison showed that peak changes from baseline aspartate aminotransferase and activated partial thromboplastin time values were significantly improved (P </=.05) with tacrolimus treatment than placebo treatment. In this prospective, double-blinded, randomized trial, we show that flushing the liver before transplantation with Plasma-lyte A containing tacrolimus results in superior early graft function and decreased hepatocellular injury after reperfusion compared with flushing with Plasma-lyte A alone.


Use of carbon mesh anodes and the effect of different pretreatment methods on power production in microbial fuel cells.
Flat electrodes are useful in microbial fuel cells (MFCs) as close electrode spacing improves power generation. Carbon cloth and carbon paper materials typically used in hydrogen fuel cells, however, are prohibitively expensive for use in MFCs. An inexpensive carbon mesh material was examined here as a substantially less expensive alternative to these materials for the anode in an MFC. Pretreatment of the carbon mesh was needed to ensure adequate MFC performance. Heating the carbon mesh in a muffle furnace (450 degrees C for 30 min) resulted in a maximum power density of 922 mW/m2 (46 W/m3) with this heat-treated anode, which was 3% more power than that produced using a mesh anode cleaned with acetone (893 mW/ m2; 45 W/m3). This power density with heating was only 7% less than that achieved with carbon cloth treated by a high temperature ammonia gas process (988 mW/m2; 49 W/m3). When the carbon mesh was treated by the ammonia gas process, power increased to 1015 mW/m2(51 W/m3). Analysis of the cleaned or heated surfaces showed these processes decreased atomic O/C ratio, indicating removal of contaminants that interfered with charge transfer. Ammonia gas treatment also increased the atomic N/C ratio, suggesting that this process produced nitrogen related functional groups that facilitated electron transfer. These results show that low cost heat-treated carbon mesh materials can be used as the anode in an MFC, providing good performance and even exceeding performance of carbon cloth anodes.


An Australasian perspective on the curative treatment of patients with pancreatic cancer, supportive care, and future directions for management.
The management of patients with pancreatic cancer requires an individualised approach and the support of a multidisciplinary team to accurately stage patients and determine their suitability for curative treatment. Guidelines have been developed in Australasia to define the operability for patients who have been diagnosed with pancreatic cancer. This is supported by advances in pancreatic cancer genetics, which show potential for developing targeted therapies for pancreatic cancer. Both surgery and targeted therapies aim to extend the overall survival of patients. Patients who are cured of their cancer may live with permanent changes in gut anatomy and physiology leading to distressing symptoms that may not be addressed. Patients who cannot be cured of pancreatic cancer may have supportive care issues that are often complex, and a strategic approach to manage these needs for patients with pancreatic cancer is underdeveloped in Australasia. Supportive care services need to be in a position to adapt patient care as the evidence base develops.


Report on a randomized trial comparing two forms of immobilization of the head for fractionated stereotactic radiotherapy.
Fractionated stereotactic radiotherapy (SRT) requires accurate and reproducible immobilization of the patient's head. This randomized study compared the efficacy of two commonly used forms of immobilization used for SRT. Two routinely used methods of immobilization, which differ in their approach to reproduce the head position from day to day, are the Gill-Thomas-Cosman (GTC) frame and the BrainLab thermoplastic mask. The GTC frame fixates on the patient's upper dentition and thus is in direct mechanical contact with the cranium. The BrainLab mask is a two-part masking system custom fitted to the front and back of the patient's head. After patients signed an IRB-approved informed consent form, eligible patients were randomized to either GTC frame or mask for their course of SRT. Patients were treated as per standard procedure; however, prior to each treatment a set of digital kilovolt images (ExacTrac, BrainLabAB, Germany) was taken. These images were fused with reference digitally reconstructed radiographs obtained from treatment planning CT to yield lateral, longitudinal, and vertical deviations of isocenter and head rotations about respective axes. The primary end point of the study was to compare the two systems with respect to mean and standard deviations using the distance to isocenter measure. A total of 84 patients were enrolled (69 patients evaluable with detailed positioning data). A mixed-effect linear regression and two-tiled t test were used to compare the distance measure for both the systems. There was a statistically significant (p < 0.001) difference between mean distances for these systems, suggesting that the GTC frame was more accurate. The mean 3D displacement and standard deviations were 3.17+1.95 mm for mask and 2.00+1.04 mm for frame. Both immobilization techniques were highly effective, but the GTC frame was more accurate. To optimize the accuracy of SRT, daily kilovolt image guidance is recommended with either immobilization system.


Lysosomal acid ceramidase ASAH1 controls the transition between invasive and proliferative phenotype in melanoma cells.
Phenotypic plasticity and subsequent generation of intratumoral heterogeneity underly key traits in malignant melanoma such as drug resistance and metastasis. Melanoma plasticity promotes a switch between proliferative and invasive phenotypes characterized by different transcriptional programs of which MITF is a critical regulator. Here, we show that the acid ceramidase ASAH1, which controls sphingolipid metabolism, acted as a rheostat of the phenotypic switch in melanoma cells. Low ASAH1 expression was associated with an invasive behavior mediated by activation of the integrin alphavbeta5-FAK signaling cascade. In line with that, human melanoma biopsies revealed heterogeneous staining of ASAH1 and low ASAH1 expression at the melanoma invasive front. We also identified ASAH1 as a new target of MITF, thereby involving MITF in the regulation of sphingolipid metabolism. Together, our findings provide new cues to the mechanisms underlying the phenotypic plasticity of melanoma cells and identify new anti-metastatic targets.


Application of neural networks to population pharmacokinetic data analysis.
This research examined the applicability of using a neural network approach to analyze population pharmacokinetic data. Such data were collected retrospectively from pediatric patients who had received tobramycin for the treatment of bacterial infection. The information collected included patient-related demographic variables (age, weight, gender, and other underlying illness), the individual's dosing regimens (dose and dosing interval), time of blood drawn, and the resulting tobramycin concentration. Neural networks were trained with this information to capture the relationships between the plasma tobramycin levels and the following factors: patient-related demographic factors, dosing regimens, and time of blood drawn. The data were also analyzed using a standard population pharmacokinetic modeling program, NON-MEM. The observed vs predicted concentration relationships obtained from the neural network approach were similar to those from NONMEM. The residuals of the predictions from neural network analyses showed a positive correlation with that from NONMEM. Average absolute errors were 33.9 and 37.3% for neural networks and 39.9% for NONMEM. Average prediction errors were found to be 2.59 and -5.01% for neural networks and 17.7% for NONMEM. We concluded that neural networks were capable of capturing the relationships between plasma drug levels and patient-related prognostic factors from routinely collected sparse within-patient pharmacokinetic data. Neural networks can therefore be considered to have potential to become a useful analytical tool for population pharmacokinetic data analysis.


2-Aminobenzoyl-CoA monooxygenase/reductase. Evidence for two distinct loci catalyzing substrate monooxygenation and hydrogenation.
2-Aminobenzoyl-CoA monooxygenase/reductase catalyzes both monooxygenation and hydrogenation of anthraniloyl-CoA. Its reactivity with 11 substrate analogs has been investigated. Only 2-aminobenzoyl-CoA (anthraniloyl-CoA) in its normal and deuterated (5-2H) form is a full substrate, and only traces of 2-hydroxybenzoyl-CoA (salicyloyl-CoA) are probably monooxygenated but not hydrogenated. The purified enzyme is a homodimer and has been resolved preparatively into three major species by anion-exchange chromatography on Mono Q. All three species have the same specific activity when reconstituted to full content of FAD, they differ, however, substantially in their mode of binding FAD. The oxidized or fully reduced enzyme forms bind tightly 0.5 mol/mol of the substrate 2-aminobenzoyl-CoA (Kd = 1-2 microM). The enzyme can be depleted of approximately 50% of its FAD, which corresponds to essentially complete removal from one of the two binding sites, reflecting a large difference in the affinity for FAD. From this it is deduced that the two sites are not equivalent. Removal of FAD from one binding site leads to loss of the hydrogenation capacity of the enzyme, while monooxygenation catalysis is retained. The FAD cofactors of the two binding sites differ drastically in their reactivities towards NADH, oxygen and N-ethylmaleimide. Exchange of reducing equivalents between the two FAD cofactors at the respective binding sites is very slow and irrelevant compared to the rates of catalysis. It is concluded that the enzyme, which has been proposed to consist of two identical polypeptide chains [Altenschmidt, U., Bokranz, M. & Fuchs, G. (1992) Eur. J. Biochem. 207, 715-722], contains two active centers which differ substantially in their catalytic activity. One center belongs to the class of monooxygenases, the other one to the (de)hydrogenases. This must result from substantially different interaction of the same flavin cofactors with protein functional groups and is, to our knowledge, unprecedented in flavoprotein enzymology.


VODKA2: A fast and accurate method to detect non-standard viral genomes from large RNA-seq datasets.
During viral replication, viruses carrying an RNA genome produce non-standard viral genomes (nsVGs), including copy-back viral genomes (cbVGs) and deletion viral genomes (delVGs), that play a crucial role in regulating viral replication and pathogenesis. Because of their critical roles in determining the outcome of RNA virus infections, the study of nsVGs has flourished in recent years exposing a need for bioinformatic tools that can accurately identify them within Next-Generation Sequencing data obtained from infected samples. Here, we present our data analysis pipeline, Viral Opensource DVG Key Algorithm2 (VODKA2), that is optimized to run on a High Performance Computing (HPC) environment for fast and accurate detection of nsVGs from large data sets.


Alleviation of hepatic steatosis accompanied by modulation of plasma and liver TNF-alpha levels by Trigonella foenum graecum (fenugreek) seeds in Zucker obese (fa/fa) rats.
Oral supplements of fenugreek (Trigonella foenum graecum) seeds (Fen) have been shown to treat glucose and lipid homeostasis in several metabolic disorders; however, its ability to alleviate obesity-associated pathologies is not known. The main objective of this study was to evaluate the effect of Fen in Zucker obese rats (Ob), an animal model of obesity and related disorders, such as dyslipidemia and hepatic steatosis. Female Zucker (6-week-old) Ob and lean (Ln) rats were randomly grouped (n = 8 rats/group) to receive either basal or 5% Fen-supplemented AIN-93G diets for 8 weeks, and then were euthanized. Histopathology and biochemical parameters in the liver together with plasma biochemistry were assessed. Obese rats had significantly higher (P < 0.05) body and liver weight, as well as plasma insulin, lactate, cholesterol, triglyceride and tumor necrosis factor (TNF)-alpha compared to their Ln counterparts. Fen significantly reduced (P < 0.05) the liver-weight of Ob rats in comparison to Ob rats fed basal diet (Control), while no effect was observed in the Ln rats. Fen treatment resulted in a lower P-value (P < 0.05). In addition, Ob rats on Fen-supplemented diets had fasting plama cholesterol and TNF-alpha levels, and significantly higher (P < 0.05) triglycerides in comparison to the control rats. Hepatic triglyceride level was significantly lower (P < 0.05) in Ob rats fed Fen supplemented diets in comparison to control. The levels of bound and soluble liver TNF-alpha (26 and 17 kDa, respectively) and TNF receptor-II (TNFR-II, 75 kDa) proteins were significantly lower (P < 0.05) in Ob than in Ln rats. Fen significantly lowered (P < 0.05) both the soluble and bound forms of TNF-alpha protein while significantly elevating (P < 0.05) TNFR-II in the livers of Ob rats compared to Ob Controls. These results demonstrate unequivocally that in a short-term preclinical evaluation, dietary Fen supplementation reduced the triglyceride accumulation in the liver, a hallmark feature of hepatic steatosis without affecting the plasma insulin or glucose levels in Zucker obese rats and suggest that TNF-alpha may play an important role in this process.


AMPK induces regulatory innate lymphoid cells after traumatic brain injury.
The CNS is regarded as an immunoprivileged organ, evading routine immune surveillance; however, the coordinated development of immune responses profoundly influences outcomes after brain injury. Innate lymphoid cells (ILCs) are cytokine-producing cells that are critical for the initiation, modulation, and resolution of inflammation, but the functional relevance and mechanistic regulation of ILCs are unexplored after acute brain injury. We demonstrate increased proliferation of all ILC subtypes within the meninges for up to 1 year after experimental traumatic brain injury (TBI) while ILCs were present within resected dura and elevated within cerebrospinal fluid (CSF) of moderate-to-severe TBI patients. In line with energetic derangements after TBI, inhibition of the metabolic regulator, AMPK, increased meningeal ILC expansion, whereas AMPK activation suppressed proinflammatory ILC1/ILC3 and increased the frequency of IL-10-expressing ILC2 after TBI. Moreover, intracisternal administration of IL-33 activated AMPK, expanded ILC2, and suppressed ILC1 and ILC3 within the meninges of WT and Rag1-/- mice, but not Rag1-/- IL2rg-/- mice. Taken together, we identify AMPK as a brake on the expansion of proinflammatory, CNS-resident ILCs after brain injury. These findings establish a mechanistic framework whereby immunometabolic modulation of ILCs may direct the specificity, timing, and magnitude of cerebral immunity.


Strategies for structural proteomics of prokaryotes: Quantifying the advantages of studying orthologous proteins and of using both NMR and X-ray crystallography approaches.
Only about half of non-membrane-bound proteins encoded by either bacterial or archaeal genomes are soluble when expressed in Escherichia coli (Yee et al., Proc Natl Acad Sci USA 2002;99:1825-1830; Christendat et al., Prog Biophys Mol Biol 200;73:339-345). This property limits genome-scale functional and structural proteomics studies, which depend on having a recombinant, soluble version of each protein. An emerging strategy to increase the probability of deriving a soluble derivative of a protein is to study different sequence homologues of the same protein, including representatives from thermophilic organisms, based on the assumption that the stability of these proteins will facilitate structural analysis. To estimate the relative merits of this strategy, we compared the recombinant expression, solubility, and suitability for structural analysis by NMR and/or X-ray crystallography for 68 pairs of homologous proteins from E. coli and Thermotoga maritima. A sample suitable for structural studies was obtained for 62 of the 68 pairs of homologs under standardized growth and purification procedures. Fourteen (eight E. coli and six T. maritima proteins) samples generated NMR spectra of a quality suitable for structure determination and 30 (14 E. coli and 16 T. maritima proteins) samples formed crystals. Only three (one E. coli and two T. maritima proteins) samples both crystallized and had excellent NMR properties. The conclusions from this work are: (1) The inclusion of even a single ortholog of a target protein increases the number of samples for structural studies almost twofold; (2) there was no clear advantage to the use of thermophilic proteins to generate samples for structural studies; and (3) for the small proteins analyzed here, the use of both NMR and crystallography approaches almost doubled the number of samples for structural studies.


A mathematical framework for analyzing T cell receptor scanning of peptides.
T cells continuously search for antigenic peptides presented on major histocompatibility complexes expressed on nearly all nucleated cells. Because only a few antigenic peptides are presented in a sea of thousands of self-peptides, the T cells have a critical task in discriminating between self- and nonself-peptides. This search process for antigens must be performed with sufficient speed in order to induce a fast response against invading pathogens. This study presents a mathematical framework for analyzing the scanning process of peptides. The framework includes analytic expressions for calculating the sampling rate as well as continuous-systems- and stochastic-agent-based models. The results show that the scanning of self-peptides is a very fast process due to fast off-rates. The simulations also predict the existence of an optimal sampling rate for a certain range of on-rates based on the recently proposed confinement time model. Calculations reveal that most of the self-peptides located within a microdomain are scanned within just a few seconds, and that the T cell receptors have kinetics for self-peptides, facilitating fast scanning. The derived mathematical expressions within this study provide conceptual calculations for further investigations of how the T cell discriminates between self- and nonself-peptides.


Do integrated mental healthcare organisations facilitate process quality in the treatment of people with schizophrenia and related psychoses?
The objective of this study is to investigate the influence of mergers of ambulatory and mental healthcare organisations on the process quality of care for persons suffering from schizophrenia or related psychoses. On the basis of the theory of Donabedian we assume the relationships between three types of quality in healthcare: structure quality, process quality and outcome quality. This study focuses on the influences of structure quality, i.e. years since merger and catchment area size upon process quality. Criteria according to Tugwell for evaluating healthcare were used to describe the process quality of schizophrenia care, resulting in a process quality questionnaire with 6 subscales and 21 items. Leading psychiatrists of 31 Dutch mental healthcare organisations, covering 89% of the country, answered the questionnaire. Both programmes and documents from the responding institutions and schizophrenia projects were analysed. Correlations of two determinants, age of the merged organisation and catchment area size, were made with total scale scores and the sub scores of the questionnaire. The response rate was 97% (31/32). Twenty-two organisations (71%) had a score of more than 50% on the used scale, 8 (29%) scored less. Two evidence-based interventions were implemented in more than 50% of the organisations, three in less than 50%. A low degree of implementation occurs in establishing care for people with schizophrenia from ethnic minorities, standardising diagnostic procedures and continuity of care. No significant relationship between the age of the merged organisation ('age') and the total process quality of schizophrenia care was found, however, the relationships between age and the subscales availability of interventions and integrated treatment were significant. No association was found between the size of the MHO's catchment area and any of the used subscales. The age of integration of residential and ambulatory mental health institutions correlates significantly with two subscales of process quality of schizophrenia care, i.e. availability of interventions and treatment. Catchment area size is not significantly associated with process quality or any of the subscales. Despite the mentioned positive effects, the overall picture of schizophrenia care is not very positive. Additional forces other than merely integration of ambulatory and residential services are needed for the further implementation of evidence-based interventions, diagnostic standards and continuity of care. The development of a national 'schizophrenia standard' (like in other countries) in relation with implementation plans and strategies to evaluate care on a regional level is recommended as well as further research on patient outcomes in relation to mergers of mental healthcare organisations.


SUDEP counseling: Where do we stand?
Sudden unexpected death in epilepsy (SUDEP) is the leading cause of epilepsy-related death in children and adults living with epilepsy. Several recent clinical practice guidelines have recommended that all individuals living with epilepsy and their caregivers be informed about SUDEP as a part of routine epilepsy counseling. Furthermore, several studies over the last two decades have explored the state of SUDEP counseling. Patients with epilepsy and their families want to be informed about the risk of SUDEP at or near the time of diagnosis, and preferably in person. Despite guideline recommendations, many pediatric and adult neurologists do not routinely inform individuals with epilepsy and their families about SUDEP. Some neurologists discuss SUDEP with only a subset of patients with epilepsy, such as those with risk factors like frequent generalized or focal to bilateral tonic-clonic seizures, nocturnal seizures, noncompliance, or medically refractory epilepsy. Proponents of routine SUDEP counseling argue that patients with epilepsy and their families have a "right to know" and that counseling may positively impact epilepsy self-management (i.e., behavioral modification and risk reduction). Some neurologists still believe that SUDEP counseling may cause unnecessary stress and anxiety for patients and their families (although this is erroneous) and that they also have a "right not to know." This narrative review explores the current gaps in SUDEP counseling, patients' and caregivers' perspectives of SUDEP counseling, and SUDEP prevention.


Tribute to R. G. Boutilier: the effect of size on the physiological and behavioural responses of oscar, Astronotus ocellatus, to hypoxia.
The physiological and behavioural responses of two size groups of oscar (Astronotus ocellatus) to hypoxia were studied. The physiological responses were tested by measuring M(O(2)) during decreasing environmental oxygen tensions. Larger oscars were better able to maintain oxygen consumption during a decrease in P(O(2)), regulating routine M(O(2)) to a significantly lower P(O(2)) threshold (50 mmHg) than smaller oscars (70 mmHg). Previous studies have also demonstrated a longer survival time of large oscars exposed to extreme hypoxia, coupled with a greater anaerobic enzymatic capability. Large oscars began aquatic surface respiration (ASR) at the oxygen tension at which the first significant decrease in M(O(2)) was seen (50 mmHg). Interestingly, smaller oscars postponed ASR to around 22 mmHg, well beyond the P(O(2)) at which they switched from oxyregulation to oxyconformation. Additionally, when given the choice between an hypoxic environment containing aquatic macrophyte shelter and an open normoxic environment, small fish showed a greater preference for the hypoxic environment. Thus shelter from predators appears particularly important for juveniles, who may accept a greater physiological compromise in exchange for safety. In response to hypoxia without available shelter, larger fish reduced their level of activity (with the exception of aggressive encounters) to aid metabolic suppression whereas smaller oscars increased their activity, with the potential benefit of finding oxygen-rich areas.


Diphenylphenoxy-Thiophene-PDI Dimers as Acceptors for OPV Applications with Open Circuit Voltage Approaching 1 Volt.
Two new perylenediimides (PDIs) have been developed for use as electron acceptors in solution-processed bulk heterojunction solar cells. The compounds were designed to exhibit maximal solubility in organic solvents, and reduced aggregation in the solid state. In order to achieve this, diphenylphenoxy groups were used to functionalize a monomeric PDI core, and two PDI dimers were bridged with either one or two thiophene units. In photovoltaic devices prepared using PDI dimers and a monomer in conjunction with PTB7, it was found that the formation of crystalline domains in either the acceptor or donor was completely suppressed. Atomic force microscopy, X-ray diffraction, charge carrier mobility measurements and recombination kinetics studies all suggest that the lack of crystallinity in the active layer induces a significant drop in electron mobility. Significant surface recombination losses associated with a lack of segregation in the material were also identified as a significant loss mechanism. Finally, the monomeric PDI was found to have sub-optimum LUMO energy matching the cathode contact, thus limiting charge carrier extraction. Despite these setbacks, all PDIs produced high open circuit voltages, reaching almost 1 V in one particular case.


High power density beam from narrow diode-laser arrays in axial symmetry.
A multiplexing method based on narrow diode-laser arrays (DLAs) in an axially symmetric configuration is described. The use of submillimeter narrow DLAs improves beam quality considerably in the slow-axis direction compared with typical 1-cm-wide DLAs. The axially symmetric geometry is advantageous for efficient spatial, wavelength, and polarization multiplexing. With narrow DLAs and an axially symmetric geometry, a small circular focus with more than 10-kW/mm2 average power density is possible. Theoretical calculations and preliminary experimental results are presented.


Toward an efficient DNAzyme.
A 33-nucleotide, guanine-rich DNA oligomer, PS5.ST1, has been reported to catalyze the metallation of mesoporphyrin IX (MPIX) by copper and zinc ions. In this paper we report a thorough investigation of the properties of this DNAzyme. We have established that a 24-nucleotide sequence (PS5.M), from within PS5.ST1, is both the minimal and most optimal catalytic unit. We have found that three related porphyrins are acceptable as substrates by this DNAzyme, of which protoporphyrin IX is preferred as a substrate over the expected substrate, MPIX. We have determined that it is unlikely that a strong, catalytically relevant binding site for copper ions exists in the DNAzyme and that high concentrations of copper destroy the active DNAzyme. This enzyme, whose folded structure likely contains guanine quartets, requires potassium ions for activity; we have shown that as little as 1 mM potassium is sufficient for its catalytic robustness, whereas as much as 0.5 M sodium still will not support catalysis. In determining the pH, temperature, and salt optima for the catalyzed reaction, we have found an unexpected stabilizing role for Tris buffer in both the catalyzed and background metallation reactions. As a consequence of various steps of optimization, we now have a vastly improved DNAzyme, one whose enzymatic parameters compare well both with those of natural ferrochelatases, as well as with those of artificially derived chelatases, composed of protein (a catalytic antibody) and RNA. The existence of this array of biocatalysts for porphyrin metallations allows one-to-one comparisons of the ways in which different biopolymers solve a given catalytic problem.


Testing a Risky Sex Behavior Intervention Pilot Website for Adolescents.
Each year, teenagers account for about one-fifth of all unintended pregnancies in the United States. As such, delivering sexual risk reduction educational materials to teens in a timely fashion is of critical importance. Web-based delivery of these materials shows promise for reaching and persuading teens away from risky sexual and substance abuse behaviors. The purpose of this study was to pilot test a web-based program aimed at reducing risky sexual behavior and related outcomes among adolescents in a high school setting. A beta-test of the website was conducted in three public schools in New Mexico, USA with 173 students in 9th and 10th grades recruited from existing health education classes. Participants spent approximately three hours over a period of two days completing the online program in school computer labs. Pretest to posttest results indicated that self-efficacy for condom use and condom use intentions, two theoretical mediators of changes in condom use behavior, were significantly changed. Adolescents also reported high satisfaction with the website content. BReady4it provided an innovative sex and substance abuse education to teenagers that revealed promising positive changes in cognitive constructs that are inversely related to risky sexual behavior among users.


Residential randon exposure and lung cancer risk in Misasa, Japan: a case-control study.
In order to investigate an association between residential radon exposure and risk of lung cancer, a case-control study was conducted in Misasa Town, Tottori Prefecture, Japan. The case series consisted of 28 people who had died of lung cancer in the years 1976-96 and 36 controls chosen randomly from the residents in 1976, matched by sex and year of birth. Individual residential radon concentrations were measured for 1 year with alpha track detectors. The average radon concentration was 46 Bq/m3 for cases and 51 Bq/m3 for controls. Compared to the level of 24 or less Bq/m3, the adjusted odds ratios of lung cancer associated with radon levels of 25-49, 50-99 and 100 or more Bq/m3, were 1.13 (95% confidence interval; 0.29-4.40), 1.23 (0.16-9.39) and 0.25 (0.03-2.33), respectively. None of the estimates showed statistical significance, due to small sample size. When the subjects were limited to only include residents of more than 30 years, the estimates did not change substantially. This study did not find that the risk pattern of lung cancer, possibly associated with residential radon exposure, in Misasa Town differed from patterns observed in other countries.


RepPop: a database for repetitive elements in Populus trichocarpa.
Populus trichocarpa is the first tree genome to be completed, and its whole genome is currently being assembled. No functional annotation about the repetitive elements in the Populus trichocarpa genome is currently available. We predicted 9,623 repetitive elements in the Populus trichocarpa genome, and assigned functions to 3,075 of them (31.95%). The 9,623 repetitive elements cover approximately 40% of the current (partially) assembled genome. Among the 9,623 repetitive elements, 668 have copies only in the contigs that have not been assigned to one of the 19 chromosome while the rest all have copies in the partially assembled chromosomes. All the predicted data are organized into an easy-to-use web-browsable database, RepPop. Various search capabilities are provided against the RepPop database. A Wiki system has been set up to facilitate functional annotation and curation of the repetitive elements by a community rather than just the database developer. The database RepPop will facilitate the assembling and functional characterization of the Populus trichocarpa genome.


22-oxa-1,25-dihydroxyvitamin D3 efficiently inhibits tumor growth in inoculated mice and primary histoculture of cholangiocarcinoma.
It is well known that 1α,25-Dihydroxyvitamin D3 (1,25[OH]2 D3) restrains cell proliferation and induces differentiation and apoptosis in normal and tumor cells. The authors of this report recently demonstrated that 1,25(OH)2 D3 effectively inhibits the proliferation of cholangiocarcinoma (CCA) cell lines. The antitumor activity and the underlying mechanism of 22-oxa-D3, an analog of vitamin D, in mice and in tissue cultures from patients with CCA were further explored in the current study. Cell growth and cell cycle distribution were examined in CCA cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Mice were injected subcutaneously with 4×10(6) CCA cells at both flank sides and intraperitoneal injections with phosphate-buffered saline or 22-oxa-D3 (15 μg/kg/day) for 17 days thereafter. Tumors were removed the next day. The expression levels of cyclin D1 and the cyclin-dependent kinase inhibitor p21 were determined by Western blot analysis and immunohistochemistry. Growth inhibition of 22-oxa-D3 in fresh tissue samples from patients with CCA was analyzed by using a histodrug response assay. 22-oxa-D3 effectively suppressed the growth of CCA cell lines in a time-dependent and dose-dependent manner. 22-oxa-D3 arrested CCA cells at G1 phase to S phase by the suppression of cyclin D1 expression and the up-regulation of p21. Supplementation of 22-oxa-D3 to CCA-inoculated mice significantly inhibited tumor growth without hypercalcemia or serious side effects. The treatment also induced cellular apoptosis in tissue samples from patients with CCA. 22-oxa-D3 effectively suppressed tumor growth in CCA-inoculated mice and induced cellular apoptosis in tissue samples from patients with CCA. The current data encourage further investigation of 1,25(OH)2 D3 or its analogues as therapeutic agents in the treatment of patients with CCA.


Surgical techniques for emergent repair of post-infarction ventricular septal defect: compare endocardial patch and infarct exclusion method with traditional method.
The traditional surgical repair of post-infarction ventricular septal defect (VSD) includes excision of necrotic myocardium and approximation of the remaining of healthy ventricular wall and septal portion. The exclusion method emphasizes no excision of infarcted myocardium, preservation of the left ventricular geometry and exclusion of infarction area. We discuss our experiences in 13 patients and compared the results obtained from 2 different surgical methods. From July 1996 to December 2001, 13 patients with post-infarction VSD received emergent repair. Seven patients were repaired in the traditional way and the other 6 with infarct exclusion method. There were 9 men and 4 women, ranging in age from 57 to 79. In the traditional group, all 7 patients were classified as NYHA IV and supported by intra-aortic balloon counter-pulsation (IABP) and 4 patients were for synchronous coronary bypass grafting. Patients using exclusion method were the 1 classified as NYHA III and 5 as IV with cardiogenic shock and supported by IABP. Coronary bypass grafting was performed concomitantly in 2 patients. Five patients died within 30 days after the surgery. Four patients (mortality rate = 57.1%) had reconstruction in traditional way and 1 (mortality rate = 16.6%) in exclusion way. The complication rate was higher in the traditional group (= 100%, n = 7, p = 0.005). In the traditional group, 1 patient received heart transplantation due to persistent severe pump failure and recovered well. Two received tracheostomy due to respiratory failure and 1 died 2 months later. In the group of exclusion method, 1 patient suffered recurrent VSD 2 days after the first surgery and died due to ventricular arrhythmia. The surgical mortality caused by acute post-infarction VSD has decreased with endocardial patch and infarction exclusion method. Rapid diagnosis, appropriate preoperative management and delicate surgical repair improve the overall results and help to attain long-term survival.


Sensory substitution: Using a vibrotactile device to orient and walk to targets.
This study investigates how active exploration helps users of sensory substitution devices (SSDs) to detect action-relevant information. A vibrotactile SSD was developed that generates stimulation that is contingent on the users' movements. Target direction was specified by the location of the vibratory stimulation, and target distance by the size and intensity of the pattern of stimulation. A series of experiments was performed with blindfolded participants. In Experiments 1a to 1c, participants used the SSD to align their central body axis with prespecified targets. These experiments differed in the number of actuators that were used and whether online perception-action coupling was present. In Experiment 2, participants approached targets with forward locomotion along a straight line. Experiment 3 combined the previous experiments and studied the concomitant walking and steering toward targets. Oscillatory movements, which facilitated information pickup, were observed in all experiments. The exploratory oscillations were shown to depend on the online perception-action coupling and were related to cases of hyperacuity, for which absolute errors were found to be smaller than the areas of sensitivity of the actuators. It is concluded that, to improve the utility of SSDs, future research with SSDs should pay more attention to the role of active information detection. (PsycINFO Database Record


Detection of biomarkers with carbon nanotube-based immunosensors.
A facile and capable method of preparation of sensitive carbon nanotube (CNT)/polysulfone/RIgG immunosensor is discussed in this chapter. The immunosensor is based on the modification of disposable screen-printed electrodes by phase inversion method. CNT/polysulfone membrane acts as the reservoir of immunomolecules as well as a transducer. This configuration offers large surface area, elevated porosity, and mechanical flexibility. The comparison with graphite/polysulfone/RIgG immunosensors shows a significantly improved sensitivity for those prepared with CNTs coupled with polysulfone (PSf). The immunosensing scheme is based on the competitive assay between free and labeled anti-RIgG for the available binding sites of immobilized rabbit IgG (RIgG). The RIgG is incorporated into the PSf immunosensor using a phase inversion method. Horseradish peroxidase enzyme is used as label and hydroquinone as electrochemical mediator. The limit of detection for competitive assay is 1.66 microg/mL. The sensitivity is six times higher for MWCNT-based than for graphite-based electrodes.


Analysis of cell-cycle kinetics and sulfur amino acid metabolism in methionine-dependent tumor cell lines; the effect of homocysteine supplementation.
Methionine dependence is a feature unique to cancer cells, exhibited as inability to grow in a methionine-depleted environment supplemented with homocysteine, the immediate metabolic precursor of methionine. This study explores the effect of methionine depletion and homocysteine supplementation on the viability, sulfur amino acid metabolism and cell-cycle kinetics of normal and cancer cells, as well as their ability to recover from the treatments. An array of cells including hepatomas (HTC, Phi-1), prostate adenocarcinomas (PC-3) and transformed (3T3) and normal (HS-27) fibroblasts, has been used aiming to evaluate the importance of tissue specificity. All cell lines proliferated well in methionine-complete media (M+H-), whilst only the normal fibroblasts HS-27 grew in methionine-depleted homocysteine-supplemented media (M-H+). None of the tested cell lines were able to grow in media without methionine or homocysteine (M-H-). HTC was the only cell line that did not recover from the M-H+ treatment whilst PC-3 did not recover from the M-H- treatment. Methionine and homocysteine depletion (M-H+ and M-H-) were found to induce arrest at different phases of the cell cycle, depending on the cell line: the methionine-dependent HTC, PC-3 and 3T3 arrested at the S and G2/M phase, whilst Phi-1 and the methionine-independent HS-27 accumulated in the G1 phase. The cell-cycle kinetics showed that the observed blockades were reversible. The information resulting from these studies is important for not only the behavior of cancer cells, but also for appreciating the potential of developing cancer therapies based on methionine-depletion strategies.


C-terminally truncated, kidney-specific variants of the WNK4 kinase lack several sites that regulate its activity.
WNK lysine-deficient protein kinase 4 (WNK4) is an important regulator of renal salt handling. Mutations in its gene cause pseudohypoaldosteronism type II, mainly arising from overactivation of the renal Na+/Cl- cotransporter (NCC). In addition to full-length WNK4, we have observed faster migrating bands (between 95 and 130 kDa) in Western blots of kidney lysates. Therefore, we hypothesized that these could correspond to uncharacterized WNK4 variants. Here, using several WNK4 antibodies and WNK4-/- mice as controls, we showed that these bands indeed correspond to short WNK4 variants that are not observed in other tissue lysates. LC-MS/MS confirmed these bands as WNK4 variants that lack C-terminal segments. In HEK293 cells, truncation of WNK4's C terminus at several positions increased its kinase activity toward Ste20-related proline/alanine-rich kinase (SPAK), unless the truncated segment included the SPAK-binding site. Of note, this gain-of-function effect was due to the loss of a protein phosphatase 1 (PP1)-binding site in WNK4. Cotransfection with PP1 resulted in WNK4 dephosphorylation, an activity that was abrogated in the PP1-binding site WNK4 mutant. The electrophoretic mobility of the in vivo short variants of renal WNK4 suggested that they lack the SPAK-binding site and thus may not behave as constitutively active kinases toward SPAK. Finally, we show that at least one of the WNK4 short variants may be produced by proteolysis involving a Zn2+-dependent metalloprotease, as recombinant full-length WNK4 was cleaved when incubated with kidney lysate.


Simulation of the Dynamics of Bacterial Quorum Sensing.
Quorum sensing (QS) is a signaling mechanism that pathogenic bacteria use to communicate and synchronize the production of exofactors to attack their hosts. Understanding and controlling QS is an important step towards a possible solution to the growing problem of antibiotic resistance. QS is a cooperative effort of a bacterial population in which some of the bacteria do not participate. This phenomenon is usually studied using game theory and the non-participating bacteria are modeled as cheaters that exploit the production of common goods (exofactors) by other bacteria. Here, we take a different approach to study the QS dynamics of a growing bacterial population. We model the bacterial population as a growing graph and use spectral graph theory to compute the evolution of its synchronizability. We also treat each bacterium as a source of signaling molecules and use the diffusion equation to compute the signaling molecule distribution. We formulate a cost function based on Lagrangian dynamics that combines the time-like synchronization with the space-like diffusion of signaling molecules. Our results show that the presence of non-participating bacteria improves the homogeneity of the signaling molecule distribution preventing thus an early onset of exofactor production and has a positive effect on the optimization of QS signaling and on attack synchronization.


Functional mapping of dorsal and median raphe 5-hydroxytryptamine pathways in forebrain of the rat using microdialysis.
Recent neurochemical studies of the properties of 5-hydroxytryptamine (5-HT) pathways arising from the dorsal raphe nucleus (DRN) and median raphe nucleus (MRN) have measured extracellular 5-HT in brain regions with reported preferential DRN or MRN 5-HT inputs. Here, we have tested whether electrical stimulation of the DRN and MRN releases 5-HT in rat forebrain regions in a pattern that fits the reported distribution of DRN/MRN pathways. The effect on extracellular 5-HT of electrical stimulation (5 Hz, 300 microA, 20 min) of the DRN, and then MRN, was determined in six regions of the anaesthetised rat. Stimulation of the DRN evoked a short-lasting but clear-cut release of 5-HT (+70-100%) in regions (frontal cortex, dorsal striatum, globus pallidus, and ventral hippocampus) reported to receive a 5-HT projection from the DRN. Regions receiving an MRN innervation (dorsal hippocampus, medial septum, and ventral hippocampus) released 5-HT (+70-100%) in response to MRN stimulation. Regions reported to receive a preferential DRN innervation (frontal cortex, dorsal striatum, and globus pallidus) did not respond to MRN stimulation. Of two regions (dorsal hippocampus and medial septum) reported to receive a preferential MRN innervation, one did not respond to DRN stimulation (dorsal hippocampus) although the other (medial septum) did. In summary, electrical stimulation of the DRN and MRN released 5-HT in a regionally specific pattern. With the exception of one region (medial septum), this pattern of release bears a strong relationship to the distribution of 5-HT projections from the DRN and MRN reported by anatomical studies. The combination of raphe stimulation with microdialysis may be a useful way to study the in vivo neurochemistry of DRN/MRN 5-HT pathways.