Vascular cognitive impairment: epidemiology, subtypes, diagnosis and management.
Dementia occurs after stroke in 25% of patients but also can arise from covert cerebrovascular disease (CVD). 'Silent' lacunes occur in 25% of the elderly, often associated with focal or confluent hyperintensities on T2-weighted magnetic resonance imaging, which are detected in 95% of seniors. These covert infarcts predict future stroke and faster cognitive decline. Best practice guidelines advocate screening for cognitive impairment in all phases of overt stroke, when covert CVD is uncovered, when vascular risk factors are present and if patients present with cognitive complaints. Standardised testing is recommended, emphasising executive function and speed of processing. Cholinesterase inhibitors have cognitive enhancing effects in vascular dementia, but the major thrust is still aggressive management of vascular risk factors and healthy lifestyle choices. Given that mixed Alzheimer's dementia and CVD is likely the most common substrate for dementia and that they share common vascular risk factors, a major goal for vascular medicine is cerebrovascular protection, not just to prevent heart attack and stroke, but also to maintain brain health and delay dementia.


Evaluation of autoregressive covariance structures for test-day records of Holstein cows: estimates of parameters.
The objective of this study was to compare test-day (TD) models with autoregressive covariance structures for the estimation of genetic and environmental components of variance for milk, fat and protein yields, and somatic cell score (SCS) in Holstein cows. Four models were compared: model I (CS model) was a simple TD repeatability animal model with compound symmetry covariance structure for environmental effects, model II (ARpe model) and model III (ARe model) had first-order autoregressive covariance structures for TD permanent or residual environmental effects, respectively, and model IV (305-d model) was a simple animal model using 305-d records. Data were 106,472 first-lactation TD records of 12,071 Holstein cows calving from 1996 through 2001. Likelihood ratio tests indicated that ARpe and ARe models fit the data significantly better than the CS model. The ARe model resulted in slightly smaller estimates of genetic variance and heritability than did the CS model. Estimates of residual variance were always smaller with the CS model than with the ARe model with the autoregressive covariance structure among TD residual effects. Estimates of heritability with different TD models were in the range of 0.06 to 0.11. The 305-d model resulted in estimates of heritability in the range of 0.11 to 0.36. The autoregressive covariance structure among TD residual effects may help to prevent bias in heritability estimates for milk, fat and protein yields, and SCS.


Requirement of Math1 for secretory cell lineage commitment in the mouse intestine.
The mouse small intestinal epithelium consists of four principal cell types deriving from one multipotent stem cell: enterocytes, goblet, enteroendocrine, and Paneth cells. Previous studies showed that Math1, a basic helix-loop-helix (bHLH) transcription factor, is expressed in the gut. We find that loss of Math1 leads to depletion of goblet, enteroendocrine, and Paneth cells without affecting enterocytes. Colocalization of Math1 with Ki-67 in some proliferating cells suggests that secretory cells (goblet, enteroendocrine, and Paneth cells) arise from a common progenitor that expresses Math1, whereas absorptive cells (enterocytes) arise from a progenitor that is Math1-independent. The continuous rapid renewal of these cells makes the intestinal epithelium a model system for the study of stem cell regeneration and lineage commitment.


Fish oil supplementation augments post-exercise immune function in young males.
Fish oils and related fatty acid components have anti-inflammatory properties, with beneficial effects against various disorders such as cardiovascular disease. A single bout of exercise can alter immune function. However, the effects of fish oil on immune function after a single bout of exercise are currently unknown. This study investigated the effect of supplementation with fish oil on the immune response to an acute bout of endurance exercise. Sixteen male subjects underwent a six week double blind randomised placebo controlled supplementation trial involving two groups (fish oil or placebo oil, 3g/day). They attended for two visits, the first involving a maximal exercise test and the second involving a 1-h bout of endurance exercise on a cycle ergometer at 70% (V)O(2peak). Blood samples were taken pre-supplementation, pre-exercise (post-supplementation), immediately, 1 and 3h post-exercise. Samples were analysed for plasma IL-6, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and cortisol; peripheral blood mononuclear cell (PBMC) IL-2, IL-4 and IFN-γ production; neutrophil phagocytosis/oxidative burst; and natural killer (NK) cell cytotoxic activity. Post-supplementation EPA concentration was increased (P=0.0127) in the fish oil group. At 3h post-exercise PBMC IL-2 (P=0.0067) and NK cell activity (P=0.0163) was greater in the fish oil compared with the control group. However, PBMC IL-4 and IFN-γ productions, plasma IL-6 and cortisol concentrations, as well as neutrophil activity were unaffected by fish oil supplementation. The current study demonstrates that fish oil supplementation reduces increases PBMC IL-2 production and NK cell cytotoxic activity in the recovery period after exercise.


Diagnostic Accuracy of a Machine-Learning Approach to Coronary Computed Tomographic Angiography-Based Fractional Flow Reserve: Result From the MACHINE Consortium.
Coronary computed tomographic angiography (CTA) is a reliable modality to detect coronary artery disease. However, CTA generally overestimates stenosis severity compared with invasive angiography, and angiographic stenosis does not necessarily imply hemodynamic relevance when fractional flow reserve (FFR) is used as reference. CTA-based FFR (CT-FFR), using computational fluid dynamics (CFD), improves the correlation with invasive FFR results but is computationally demanding. More recently, a new machine-learning (ML) CT-FFR algorithm has been developed based on a deep learning model, which can be performed on a regular workstation. In this large multicenter cohort, the diagnostic performance ML-based CT-FFR was compared with CTA and CFD-based CT-FFR for detection of functionally obstructive coronary artery disease. At 5 centers in Europe, Asia, and the United States, 351 patients, including 525 vessels with invasive FFR comparison, were included. ML-based and CFD-based CT-FFR were performed on the CTA data, and diagnostic performance was evaluated using invasive FFR as reference. Correlation between ML-based and CFD-based CT-FFR was excellent (R=0.997). ML-based (area under curve, 0.84) and CFD-based CT-FFR (0.84) outperformed visual CTA (0.69; P<0.0001). On a per-vessel basis, diagnostic accuracy improved from 58% (95% confidence interval, 54%-63%) by CTA to 78% (75%-82%) by ML-based CT-FFR. The per-patient accuracy improved from 71% (66%-76%) by CTA to 85% (81%-89%) by adding ML-based CT-FFR as 62 of 85 (73%) false-positive CTA results could be correctly reclassified by adding ML-based CT-FFR. On-site CT-FFR based on ML improves the performance of CTA by correctly reclassifying hemodynamically nonsignificant stenosis and performs equally well as CFD-based CT-FFR.


Is it worth treating fever in intensive care unit patients? Preliminary results from a randomized trial of the effect of external cooling.
Antipyresis is a common clinical practice in intensive care, although it is unknown if fever is harmful, beneficial, or a negligible adverse effect of infection and inflammation. In a randomized study, rectal temperature and discomfort were assessed in 38 surgical intensive care unit patients without neurotrauma or severe hypoxemia and with fever (temperature >/=38.5 degrees C) and systemic inflammatory response syndrome. Eighteen patients received external cooling while 20 received no antipyretic treatment. Temperature and discomfort decreased similarly in both groups after 24 hours. No significant differences in recurrence of fever, incidence of infection, antibiotic therapy, intensive care unit and hospital length of stay, or mortality were noted between the groups. These results suggest that the systematic suppression of fever may not be useful in patients without severe cranial trauma or significant hypoxemia. Letting fever take its natural course does not seem to harm patients with systemic inflammatory response syndrome or influence the discomfort level and may save costs.


Control of gaze while walking: Task structure, reward, and uncertainty.
While it is universally acknowledged that both bottom up and top down factors contribute to allocation of gaze, we currently have limited understanding of how top-down factors determine gaze choices in the context of ongoing natural behavior. One purely top-down model by Sprague, Ballard, and Robinson (2007) suggests that natural behaviors can be understood in terms of simple component behaviors, or modules, that are executed according to their reward value, with gaze targets chosen in order to reduce uncertainty about the particular world state needed to execute those behaviors. We explore the plausibility of the central claims of this approach in the context of a task where subjects walk through a virtual environment performing interceptions, avoidance, and path following. Many aspects of both walking direction choices and gaze allocation are consistent with this approach. Subjects use gaze to reduce uncertainty for task-relevant information that is used to inform action choices. Notably the addition of motion to peripheral objects did not affect fixations when the objects were irrelevant to the task, suggesting that stimulus saliency was not a major factor in gaze allocation. The modular approach of independent component behaviors is consistent with the main aspects of performance, but there were a number of deviations suggesting that modules interact. Thus the model forms a useful, but incomplete, starting point for understanding top-down factors in active behavior.


Hydrodynamics strongly affect the dynamics of colloidal gelation but not gel structure.
Colloidal particles with strong, short-ranged attractions can form a gel. We simulate this process without and with hydrodynamic interactions (HI), using the lattice-Boltzmann method to account for presence of a thermalized solvent. We show that HI speed up and slow down gelation at low and high volume fractions, respectively. The transition between these two regimes is linked to the existence of a percolating cluster shortly after quenching the system. However, when we compare gels at matched 'structural age', we find nearly indistinguishable structures with and without HI. Our result explains longstanding, unresolved conflicts in the literature.


Investigation of immune and CNS-mediated effects of fingolimod in the focal delayed-type hypersensitivity multiple sclerosis model.
We examined the effect of fingolimod (0.1 and 0.3 mg/kg/day orally) on blood-brain barrier (BBB) function, demyelination and leukocyte recruitment at different stages of the focal delayed-type hypersensitivity (DTH) multiple sclerosis model in Lewis rats using immunohistochemistry and gadolinium (Gd)-enhancing magnetic resonance imaging (MRI). During DTH lesion formation, fingolimod reduced BBB breakdown (52%; p = 0.05), and lymphocyte (53%; p = 0.016) and macrophage/activated microglia (49%; p = 0.002) recruitment to the DTH lesion compared with vehicle-treated controls. Following DTH lesion establishment, fingolimod reduced the area of BBB breakdown (75%; p = 0.04), lymphocyte recruitment to the DTH lesion (41%; p = 0.01) and activated microglia outside of the lesion core (p = 0.01), but did not reduce recruitment of macrophages/activated microglia within the DTH lesion. During the chronic disease phase, when the BBB was resealed, fingolimod reduced the area of demyelination by 43% (p = 0.019) compared with vehicle-treated controls, while not affecting lymphocyte recruitment within the lesion. Fingolimod had different beneficial effects during different stages of DTH, reducing BBB breakdown and lesion development/brain tissue damage whilst reducing lymphocyte recruitment when BBB breakdown was apparent, but reducing demyelination independent of lymphocyte infiltration behind an intact BBB. These results suggest a direct CNS effect of fingolimod in this model.


In vivo imaging of leucine aminopeptidase activity in drug-induced liver injury and liver cancer via a near-infrared fluorescent probe.
The liver, a main detoxification organ, has evolved a complex enzymatic system to respond to multiple pathological conditions, in which leucine aminopeptidase (LAP) has been reported to participate in detoxifying cisplatin in hepatoma cells and contribute to the intrinsic drug resistance. In vivo imaging of LAP activity in liver disease models is thus helpful to further understand the function of LAP in detoxification and medicine, but such an imaging approach is still lacking. Herein, we develop a selective and sensitive near-infrared fluorescent probe (HCAL) for this purpose. Using the probe, combined with confocal fluorescence imaging, we disclose the upregulations of LAP in acetaminophen-induced liver injury and tumor-bearing mice models. Supplementary acetylcysteine can suppress this upregulation, revealing that the LAP increase may be connected with a deficiency in biothiols. Moreover, HCAL has been used to image LAP in hepatoma cells, tumor tissues and xenograft tumor mice models successfully. These results demonstrate that HCAL may be a promising tool for studying the function of LAP in LAP-associated liver diseases.


Natural and traditional medicine in cuba: lessons for u.s. Medical education.
Reprinted with permission from Academic Medicine (Academic Medicine. 81(12):1098-1103, December 2006). The Institute of Medicine's (IOM's) Academy of Science has recommended that medical schools incorporate information on CAM (complementary and alternative medicine) into required medical school curricula so that graduates will be able to competently advise their patients in the use of CAM. The report states a need to study models of systems that integrate CAM and allopathic medicine. The authors present Cuba's health care system as one such model and describe how CAM (or natural and traditional medicine) is integrated into all levels of clinical care and medical education in Cuba. The authors examine the Cuban medical school curriculum in which students, residents, and practicing physicians are oriented in the two paradigms of CAM and allopathic medicine. Only health professionals are permitted to practice CAM in Cuba; therefore, Cuba's medical education curriculum incorporates not only teaching about CAM, but it also teaches basic CAM approaches and clinical skills. Both the theory and practice of CAM are integrated into courses throughout the six-year curriculum. Similarities and differences between the U.S. and Cuban approaches to CAM are examined, including issues of access and cost, and levels of acceptance by the medical profession and by the public at large in both countries. The authors conclude that there is potentially much to learn from the Cuban experience to inform U.S. medical educators and institutions in their endeavors to comply with the IOM recommendations and to incorporate CAM into medical school curricula. Acad Med. 2006; 81:1098-1103.


Anion binding by human lactoferrin: results from crystallographic and physicochemical studies.
The anion-binding properties of lactoferrin (Lf), with Fe3+ or Cu2+ as the associated metal ion, have been investigated by physicochemical and crystallographic techniques. These highlight differences between the two sites and in the anion-binding behavior when different metals are bound. Carbonate, oxalate, and hybrid carbonate-oxalate complexes have been prepared and their characteristic electronic and EPR spectra recorded. Oxalate can displace carbonate from either one or both anion sites of Cu2(CO3)2Lf, depending on the oxalate concentration, but no such displacement occurs for Fe2(CO3)2Lf. Addition of oxalate and the appropriate metal ion to apoLf under carbonate-free conditions gives dioxalate complexes with both Fe3+ and Cu2+, except when traces of EDTA remain associated with the protein, when hybrid complexes M2(CO3)(C2O4)Lf can result. The anion sites in the crystal structures of Fe2(CO3)2Lf, Cu2-(CO3)2Lf, and Cu2(CO3)(C2O4)Lf, refined at 2.2, 2.1, and 2.2 A, respectively, have been compared. In every case, the anion is hydrogen bonded to the N-terminus of helix 5, an associated arginine side chain, and a nearby threonine side chain. The carbonate ion binds in bidentate fashion to the metal, except in the N-lobe site of dicupric lactoferrin, where it is monodentate; the difference arises from slight movement of the metal ion. The hybrid complex shows that the oxalate ion binds preferentially in the C-lobe site, in 1,2-bidentate mode, but with the displacement of several nearby side chains. These observations lead to a generalized model for synergistic anion binding by transferrins.


Activation of an AP1-like transcription factor of the maize pathogen Cochliobolus heterostrophus in response to oxidative stress and plant signals.
Redox sensing is a ubiquitous mechanism regulating cellular activity. Fungal pathogens face reactive oxygen species produced by the host plant's oxidative burst in addition to endogenous reactive oxygen species produced during aerobic metabolism. An array of preformed and induced detoxifying enzymes, including superoxide dismutase, catalases, and peroxidases, could allow fungi to infect plants despite the oxidative burst. We isolated a gene (CHAP1) encoding a redox-regulated transcription factor in Cochliobolus heterostrophus, a fungal pathogen of maize. CHAP1 is a bZIP protein that possesses two cysteine-rich domains structurally and functionally related to Saccharomyces cerevisiae YAP1. Deletion of CHAP1 in C. heterostrophus resulted in decreased resistance to oxidative stress caused by hydrogen peroxide and menadione, but the virulence of chap1 mutants was unaffected. Upon activation by oxidizing agents or plant signals, a green fluorescent protein (GFP)-CHAP1 fusion protein became localized in the nucleus. Expression of genes encoding antioxidant proteins was induced in the wild type but not in chap1 mutants. Activation of CHAP1 occurred from the earliest stage of plant infection, in conidial germ tubes on the leaf surface, and persisted during infection. Late in the course of infection, after extensive necrotic lesions were formed, GFP-CHAP1 redistributed to the cytosol in hyphae growing on the leaf surface. Localization of CHAP1 to the nucleus may, through changes in the redox state of the cell, provide a mechanism linking extracellular cues to transcriptional regulation during the plant-pathogen interaction.


Neuroimaging the Menstrual Cycle and Premenstrual Dysphoric Disorder.
Knowledge of gonadal hormone-related influences on human brain anatomy, function, and chemistry is scarce. The present review scrutinized organizational and functional neuroimaging correlates of the menstrual cycle and premenstrual dysphoric disorder (PMDD). Supportive evidence of cyclic short-term structural and functional brain plasticity in response to gonadal hormonal modulation is provided. The paucity of studies, sparsity and discordance of findings, and weaknesses in study design at present hinder the drawing of firm conclusions. Ideal study designs should comprise high-resolution multimodal neuroimaging (e.g., MRI, DTI, rs-fMRI, fMRI, PET), hormones, genetic, and behavioral longitudinal assessments of healthy women and PMDD patients at critical time points of the menstrual cycle phase (i.e., early follicular phase, late follicular phase, mid-luteal phase) in a counter-balanced setup. Studies integrating large-scale brain network structural, functional, and molecular neuroimaging, as well as treatment data, will deepen the understanding of neural state, disorder, and treatment markers.


Characterization of human skeletal muscle fibres according to the myosin heavy chains they express.
Using a method of single muscle fibre analysis, we investigated the presence of RNA transcripts for various isoforms of the myosin heavy chain (MyoHC) gene in histochemically, immunohistochemically and electrophoretically characterized individual muscle fibres (n = 65) from adult human vastus lateralis muscle. A cDNA clone isolated in this study was shown to contain the 3' end of a previously uncharacterized human MyoHC gene which is expressed specifically in human fast IIA muscle fibres and we conclude that this clone contains part of the human fast IIA MyoHC gene. In all the fibres histochemically, immunohistochemically and electrophoretically characterized as containing the previously classified IIB MyoHC (n = 23), it was shown that the human equivalent to the rat type IIX MyoHC gene is expressed. This observation was taken to suggest that the previously classified IIB muscles fibres in human muscle express a MyoHC isoform equivalent to the rat IIX, not the IIB, and would therefore be more accurately classified as IIX fibres.


Potent inhibitors of the Plasmodium falciparum enzymes plasmepsin I and II devoid of cathepsin D inhibitory activity.
The hemoglobin-degrading aspartic proteases plasmepsin I (Plm I) and plasmepsin II (Plm II) of the malaria parasite Plasmodium falciparum have lately emerged as putative drug targets. A series of C(2)-symmetric compounds encompassing the 1,2-dihydroxyethylene scaffold and a variety of elongated P1/P1' side chains were synthesized via microwave-assisted palladium-catalyzed coupling reactions. Binding affinity calculations with the linear interaction energy method and molecular dynamics simulations reproduced the experimental binding data obtained in a Plm II assay with very good accuracy. Bioactive conformations of the elongated P1/P1' chains were predicted and agreed essentially with a recent X-ray structure. The compounds exhibited picomolar to nanomolar inhibition constants for the plasmepsins and no measurable affinity to the human enzyme cathepsin D. Some of the compounds also demonstrated significant inhibition of parasite growth in cell culture. To the best of our knowledge, these plasmepsin inhibitors represent the most selective reported to date and constitute promising lead compounds for further optimization.


Selection and evaluation of housekeeping genes for haemocytes of soft-shell clams (Mya arenaria) challenged with Vibrio splendidus.
Gene expression studies have opened a tremendous field of investigation in biological research over the last decades. Expression of genes is most frequently quantified by real time PCR (RT-qPCR), as this method has proven to be highly sensitive. One of the critical steps, however, in comparing transcription profiles is the availability of selected housekeeping genes. Expression of these genes should be steadily stable across the conditions under study so that they provide a baseline for gene expression comparison. Such a baseline is best established using a set of few housekeeping genes. Usually, those genes are involved in maintaining homeostasis and cell viability. In our study, nine candidate genes were used, including some commonly used housekeeping genes, such as ribosomal RNA (18S, S-15, S-18 and L-37), beta actin, ubiquitin, receptor activated C kinase (RACK) and elongation factor 1 and 2, in order to determine the most stable housekeeping genes, after haemocytes of Mya arenaria were exposed to Vibrio splendidus for 2 h. Our results showed that EF-1, S-18 and ubiquitin appear to be the most stable genes for this experimental condition. On the other hand, both 18S and beta actin, the most widely used housekeeping genes, turned out to be the least stable. This demonstrates the absolute need for preliminary assessment of housekeeping genes in gene expression studies.


NEP-TC a rRNA Methyltransferase Involved on Somatic Embryogenesis of Tamarillo (Solanum betaceum Cav.).
Somatic embryogenesis (SE) is an important biotechnological tool for large-scale clonal propagation and for embryogenesis research. Moreover, genetic transformation and cryopreservation procedures in many species rely on efficient SE protocols. We have been studying different aspects related to SE induction and somatic embryo development in tamarillo (Solanum betaceum Cav.), a small tree from the Solanaceae family. Previous proteomic analyses identified a protein (NEP-TC, 26.5 kDa) consistently present in non-embryogenic calluses of tamarillo, but absent in the embryogenic ones. In this work, the role of NEP-TC during SE was assessed by gene expression analysis and immunolocalization. The results obtained demonstrated that NEP-TC is a putative member of the SpoU rRNA methylase family. This protein, present in the cytoplasm and nucleus, is expressed in non-embryogenic cells and not expressed in embryogenic cells. Slightly enhanced SE induction levels in tamarillo plants with NEP-TC down-regulated levels also supports the role of this protein on SE induction. Heterologous expression was used to confirm NEP-TC rRNA methyltransferase activity, with enhanced activity levels when rRNA was used as a substrate. These data relate a putative member of the SpoU methylase family with plant morphogenesis, in particular with SE induction.


Reducing a spatial database to its effective dimensionality for logistic-regression analysis of incidence of livestock disease.
Large databases with multiple variables, selected because they are available and might provide an insight into establishing causal relationships, are often difficult to analyse and interpret because of multicollinearity. The objective of this study was to reduce the dimensionality of a multivariable spatial database of Zimbabwe, containing many environmental variables that were collected to predict the distribution of outbreaks of theileriosis (the tick-borne infection of cattle caused by Theileria parva and transmitted by the brown ear tick). Principal-component analysis and varimax rotation of the principal components were first used to select a reduced number of variables. The logistic-regression model was evaluated by appropriate goodness-of-fit tests.


Socioeconomic burden of psoriatic arthritis in Hong Kong: direct and indirect costs and the influence of disease pattern.
To estimate the direct costs and indirect costs of patients with psoriatic arthritis (PsA) in Hong Kong. A retrospective cost-of-illness study was performed on 125 patients with PsA. Participants completed questionnaires on demographics, employment status, and out of pocket expenses. Health resources consumption was recorded by chart review and patient self-report questionnaire. Patients were grouped according to disease pattern, i.e., peripheral and axial disease. Multiple regression was used to determine the predictors of the costs. The average annual direct costs were $4,141 (2006 US dollars) per patient. Costs of inpatient care accounted for 27% of direct costs, followed by costs of visits to healthcare providers (25%). The estimated average indirect costs were $3,127 per patient-year. Forty-eight (42%) patients had no indirect costs. Sixty percent of patients with peripheral disease were still employed, compared to 39% of patients with axial disease. Patients with axial disease had almost twice the indirect costs compared to those with peripheral disease (p = 0.005). Increased pain and poor function were independently associated with increased direct costs. Worse physical health status, determined by indirect costs borne by the patient, and poor function and old age predicted high costs. PsA imposes substantial economic burden. Pain and function are significantly associated with costs. Improvements in treatments to reduce pain and restore function are likely to reduce the costs incurred by these patients.


Three-dimensional organization of otolith-ocular reflexes in rhesus monkeys. II. Inertial detection of angular velocity.
1. The dynamic contribution of otolith signals to three-dimensional angular vestibuloocular reflex (VOR) was studied during off-vertical axis rotations in rhesus monkeys. In an attempt to separate response components to head velocity from those to head position relative to gravity during low-frequency sinusoidal oscillations, large oscillation amplitudes were chosen such that peak-to-peak head displacements exceeded 360 degrees. Because the waveforms of head position and velocity differed in shape and frequency content, the particular head position and angular velocity sensitivity of otolith-ocular responses could be independently assessed. 2. During both constant velocity rotation and low-frequency sinusoidal oscillations, the otolith system generated two different types of oculomotor responses: 1) modulation of three-dimensional eye position and/or eye velocity as a function of head position relative to gravity, as presented in the preceding paper, and 2) slow-phase eye velocity as a function of head angular velocity. These two types of otolith-ocular responses have been analyzed separately. In this paper we focus on the angular velocity responses of the otolith system. 3. During constant velocity off-vertical axis rotations, a steady-state nystagmus was elicited that was maintained throughout rotation. During low-frequency sinusoidal off-vertical axis oscillations, dynamic otolith stimulation resulted primarily in a reduction of phase leads that characterize low-frequency VOR during earth-vertical axis rotations. Both of these effects are the result of an internally generated head angular velocity signal of otolithic origin that is coupled through a low-pass filter to the VOR. No change in either VOR gain or phase was observed at stimulus frequencies larger than 0.1 Hz. 4. The dynamic otolith contribution to low-frequency angular VOR exhibited three-dimensional response characteristics with some quantitative differences in the different response components. For horizontal VOR, the amplitude of the steady-state slow-phase velocity during constant velocity rotation and the reduction of phase leads during sinusoidal oscillation were relatively independent of tilt angle (for angles larger than approximately 10 degrees). For vertical and torsional VOR, the amplitude of steady-state slow-phase eye velocity during constant velocity rotation increased, and the phase leads during sinusoidal oscillation decreased with increasing tilt angle. The largest steady-state response amplitudes and smallest phase leads were observed during vertical/torsional VOR about an earth-horizontal axis. 5. The dynamic range of otolith-borne head angular velocity information in the VOR was limited to velocities up to approximately 110 degrees/s. Higher head velocities resulted in saturation and a decrease in the amplitude of the steady-state response components during constant velocity rotation and in increased phase leads during sinusoidal oscillations. 6. The response characteristics of otolith-borne angular VORs were also studied in animals after selective semicircular canal inactivation. Otolith angular VORs exhibited clear low-pass filtered properties with a corner frequency of approximately 0.05-0.1 Hz. Vectorial summation of canal VOR alone (elicited during earth-vertical axis rotations) and otolith VOR alone (elicited during off-vertical axis oscillations after semicircular canal inactivation) could not predict VOR gain and phase during off-vertical axis rotations in intact animals. This suggests a more complex interaction of semicircular canal and otolith signals. 7. The results of this study show that the primate low-frequency enhancement of VOR dynamics during off-vertical axis rotation is independent of a simultaneous activation of the vertical and torsional "tilt" otolith-ocular reflexes that have been characterized in the preceding paper. (ABSTRACT TRUNCATED)


Autophosphorylation of the Bacterial Tyrosine-Kinase CpsD Connects Capsule Synthesis with the Cell Cycle in Streptococcus pneumoniae.
Bacterial capsular polysaccharides (CPS) are produced by a multi-protein membrane complex, in which a particular type of tyrosine-autokinases named BY-kinases, regulate their polymerization and export. However, our understanding of the role of BY-kinases in these processes remains incomplete. In the human pathogen Streptococcus pneumoniae, the BY-kinase CpsD localizes at the division site and participates in the proper assembly of the capsule. In this study, we show that the cytoplasmic C-terminal end of the transmembrane protein CpsC is required for CpsD autophosphorylation and localization at mid-cell. Importantly, we demonstrate that the CpsC/CpsD complex captures the polysaccharide polymerase CpsH at the division site. Together with the finding that capsule is not produced at the division site in cpsD and cpsC mutants, these data show that CPS production occurs exclusively at mid-cell and is tightly dependent on CpsD interaction with CpsC. Next, we have analyzed the impact of CpsD phosphorylation on CPS production. We show that dephosphorylation of CpsD induces defective capsule production at the septum together with aberrant cell elongation and nucleoid defects. We observe that the cell division protein FtsZ assembles and localizes properly although cell constriction is impaired. DAPI staining together with localization of the histone-like protein HlpA further show that chromosome replication and/or segregation is defective suggesting that CpsD autophosphorylation interferes with these processes thus resulting in cell constriction defects and cell elongation. We show that CpsD shares structural homology with ParA-like ATPases and that it interacts with the chromosome partitioning protein ParB. Total internal reflection fluorescence microscopy imaging demonstrates that CpsD phosphorylation modulates the mobility of ParB. These data support a model in which phosphorylation of CpsD acts as a signaling system coordinating CPS synthesis with chromosome segregation to ensure that daughter cells are properly wrapped in CPS.


Moderate and high intensity sprint exercise induce differential responses in COX4I2 and PDK4 gene expression in Thoroughbred horse skeletal muscle.
The role of molecular signalling pathways in the phenotypic adaptation of skeletal muscle to different exercise stimuli in the Thoroughbred horse has not been reported previously. To examine CKM, COX4I1, COX4I2 and PDK4 gene expression following high intensity sprint and moderate intensity treadmill exercise stimuli in skeletal muscle of Thoroughbred horses. Two groups of trained 3-year-old Thoroughbred horses participated. Group A (n = 6 females, n = 3 males) participated in an incremental step test (moderate intensity) to fatigue or HR(max) on a Sato high speed treadmill (distance = 5418.67 m ± 343.21). Group B (n = 8 females) participated in routine 'work' (sprint) on an all-weather gallop (distance = 812.83 m ± 12.53). Biopsy samples were obtained from the gluteus medius pre-exercise (T(0)), immediately post exercise (T(1)) and 4 h post exercise (T(2)). For physiological relevance venous blood samples were collected to measure plasma lactate and creatine kinase concentrations. Changes in mRNA expression were determined by quantitative real-time RT-PCR for creatine kinase muscle (CKM), cytochrome c oxidase subunit IV isoform 1 (COX4I1), cytochrome c oxidase subunit IV isoform 2 (COX4I2) and pyruvate dehydrogenase kinase, isozyme 4 (PDK4) genes. Statistical significance (α < 0.05) was determined using Student's t tests. COX4I2 mRNA expression decreased significantly in Group A and remained unchanged in Group B between T(0) vs. T(2) (-1.7-fold, P = 0.017; -1.0-fold, P = 0.859). PDK4 mRNA expression increased significantly in Group B but not in Group A between T(0) vs. T(1) (3.8-fold, P = 0.039; 1.4-fold, P = 0.591). There were no significant changes in the expression in CKM or COX4I1 mRNA abundance in either group. Different exercise protocols elicit variable transcriptional responses in key exercise relevant genes in equine skeletal muscle due to variation in metabolic demand.


Sevoflurane Induces Learning and Memory Impairment in Young Mice Through a Reduction in Neuronal Glucose Transporter 3.
Sevoflurane, which is widely used in paediatric anaesthesia, induces neural apoptosis in the developing brain and cognitive impairment in young mammals. Glucose hypometabolism is the key pathophysiological modulator of cognitive dysfunction. However, the effects and mechanism of sevoflurane on cerebral glucose metabolism after its use as an anaesthetic and its complete elimination are still unknown. We therefore investigated the influence of sevoflurane on neuronal glucose transporter isoform 3 (GLUT3) expression, glucose metabolism and apoptosis in vivo and in vitro and on neurocognitive function in young mice 24 h after the third exposure to sevoflurane. Postnatal day 14 (P14) mice and neural cells were exposed to 3% sevoflurane 2 h daily for three days. We found that sevoflurane anaesthesia decreased GLUT3 gene and protein expression in the hippocampus and temporal lobe, consistent with a decrease in glucose metabolism in the hippocampus and temporal lobe observed by [18F] fluorodeoxyglucose positron emission tomography (18F-FDG PET). Moreover, sevoflurane anaesthesia increased the number of TUNEL-positive cells and the levels of Bax, cleaved caspase 3 and cleaved PARP and reduced Bcl-2 levels in the hippocampus and temporal lobe. Young mice exposed to sevoflurane multiple times also showed learning and memory impairment. In addition, sevoflurane inhibited GLUT3 expression in primary hippocampal neurons and PC12 cells. GLUT3 overexpression in cultured neurons ameliorated the sevoflurane-induced decrease in glucose utilization and increase in the apoptosis rate. These data indicate that GLUT3 deficiency may contribute to sevoflurane-induced learning and memory deficits in young mice.


Long term changes in human corneal endothelium following toxic endothelial cell destruction: a specular microscopic and fluorophotometric study.
To investigate the long term relation between corneal thickness, endothelial morphometric variables, and endothelial permeability in patients with endothelial cell counts under 900 cells/mm2 as a result of endothelial cell destruction after cataract surgery. Eighteen patients developed the so called toxic endothelial cell destruction (TECD) syndrome following routine cataract surgery because of the intracameral injection of a toxic detergent residue. Ten patients with a mean (SEM) initial cell loss of 72% (2%) were followed for 4 years. Data were obtained at 6 months and 4 years postoperatively and compared between TECD eyes and contralateral control eyes. Mean (SEM) endothelial cell density of the TECD eyes increased from 642 (41) cells/mm2 to 849 (50) cells/mm2 at 4 years postoperatively (p = 0.005). There was no difference in coefficient of variation or percentage hexagonals between 6 months and 4 years postoperatively. Mean (SD) corneal thickness of the TECD eyes and control eyes was similar, 0.51 (0.02) mm and 0.49 (0.01) mm, respectively (p = 0.65). Mean (SD) endothelial permeability was also similar for TECD eyes and control eyes (4.3 (0.9) x 10(-4) cm/min and 4.4 (0.6) x 10(-4) cm/min, respectively (p = 0.57). There was no correlation between endothelial cell density, coefficient of variation, or percentage of hexagonal cells and endothelial permeability in the TECD eyes. In three patients a permanent corneal decompensation occurred. Four years after TECD corneal endothelial wound healing is stable and the barrier function has been restored.


Maternal depressive symptoms and child behavior problems: Attachment security as a protective factor.
Maternal depressive symptoms (MDS) have been linked to both child internalizing and externalizing behavior problems. Theory suggests that child attachment security may be a protective factor against the negative effects of MDS. This study examined child attachment security as a buffer of the link between MDS and child internalizing and externalizing behavior problems at two time points in a predominantly African American sample. Participants included mothers (N = 164; Mage = 29.68 years; 76% African American) and their preschool-aged children (60% girls; Mage = 44.67 months) recruited from four Head Start centers in low-income neighborhoods in Baltimore, Maryland. MDS were concurrently associated with child internalizing and externalizing behavior problems at both time points. No significant main effects of child attachment security on behavior problems emerged; however, child attachment moderated the association between MDS and child internalizing behavior problems at Time 2, such that MDS predicted greater child internalizing problems when attachment security was low, and the effect was attenuated when attachment security was high. No interaction emerged for child externalizing problems. Findings suggest that secure attachment in early childhood can serve as a protective factor in the context of parental risk. We discuss implications for intervention and the intergenerational transmission of psychopathology.


Recent developments of ketene dithioacetal chemistry.
Ketene dithioacetals are versatile intermediates in organic synthesis. Extensive research, since the last decade, has given rise to new prospects in their chemistry. The objective of this review is twofold: first, to highlight the new prospects in the chemistry of functionalized ketene dithioacetals, and second, to provide an intrinsic link between ketene dithioacetal groups and a variety of other functional groups, which has brought out many new facts that will assist in future designs.


Assessment of GFP fluorescence in cells of Streptococcus gordonii under conditions of low pH and low oxygen concentration.
Use of green fluorescent protein (GFP) as a molecular reporter is restricted by several environmental factors, such as its requirement for oxygen in the development of the fluorophore, and its poor fluorescence at low pH. There are conflicting data on these limitations, however, and systematic studies to assess the importance of these factors for growing bacterial cultures are lacking. In the present study, homogeneous expression of the gfpmut3* gene directed by a synthetic constitutive lactococcal promoter was demonstrated in batch cultures and in biofilms of Streptococcus gordonii DL1. A lower limit of oxygen concentration for maturation of the GFP fluorophore was determined: fluorescence was emitted at 0.1 p.p.m. dissolved oxygen (in conventionally prepared anaerobic media lacking reducing agents), whereas no fluorescence was detected in the presence of 0.025 p.p.m. dissolved oxygen (obtained by addition of L-cysteine as reducing agent). When an anaerobically grown (non-fluorescent) >50 microm thick biofilm was shifted to aerobic conditions, fluorescence could be detected within 4 min, reaching a maximum over the next 16 min. It was not possible to detect any fluorescence gradients (lateral or vertical) within the >50 microm thick biofilm, and fluorescence development after the shift to aerobic conditions occurred throughout the biofilm (even at the substratum). This suggests that oxygen gradients, which might result in reduced GFP fluorescence, did not exist in the >50 microm thick biofilm of this organism. Production of lactic acid and the subsequent acidification in batch cultures of S. gordonii DL1 led to a decrease in fluorescence intensity. However, severe pH reduction was prevented when the bacterium was grown as a biofilm in a flowcell, and a homogeneous distribution of a strong fluorescence signal was observed. These findings show that GFP can be applied to studies of oxygen-tolerant anaerobic bacteria, that densely packed, flowcell-grown biofilms of S. gordonii do not develop oxygen gradients inhibitory to GFP fluorescence development, and that the often transient nature of GFP fluorescence in acid-producing bacteria can be overcome in flowcells, probably by the elimination of metabolic by-product accumulation.


Listening beyond seeing: Event-related potentials to audiovisual processing in visual narrative.
Every day we integrate meaningful information coming from different sensory modalities, and previous work has debated whether conceptual knowledge is represented in modality-specific neural stores specialized for specific types of information, and/or in an amodal, shared system. In the current study, we investigated semantic processing through a cross-modal paradigm which asked whether auditory semantic processing could be modulated by the constraints of context built up across a meaningful visual narrative sequence. We recorded event-related brain potentials (ERPs) to auditory words and sounds associated to events in visual narratives-i.e., seeing images of someone spitting while hearing either a word (Spitting!) or a sound (the sound of spitting)-which were either semantically congruent or incongruent with the climactic visual event. Our results showed that both incongruent sounds and words evoked an N400 effect, however, the distribution of the N400 effect to words (centro-parietal) differed from that of sounds (frontal). In addition, words had an earlier latency N400 than sounds. Despite these differences, a sustained late frontal negativity followed the N400s and did not differ between modalities. These results support the idea that semantic memory balances a distributed cortical network accessible from multiple modalities, yet also engages amodal processing insensitive to specific modalities.


Genetic diversity of Dekkera bruxellensis yeasts isolated from Australian wineries.
Yeasts of the genus Dekkera and its anamorph Brettanomyces represent a significant spoilage issue for the global wine industry. Despite this, there is limited knowledge of genetic diversity and strain distribution within wine and winery-related environments. In this study, amplified fragment length polymorphism (AFLP) analysis was conducted on 244 Dekkera bruxellensis isolates from red wine made in 31 winemaking regions of Australia. The results indicated there were eight genotypes among the isolates, and three of these were commonly found across multiple winemaking regions. Analysis of 26S rRNA gene sequences provided further evidence of three common, conserved groups, whereas a phylogeny based upon the AFLP data demonstrated that the most common D. bruxellensis genotype (I) in Australian red wine was highly divergent from the D. bruxellensis type strain (CBS 74).


Variations in the expression of pili: the effect on adherence of Neisseria meningitidis to human epithelial and endothelial cells.
The effect of variations in Neisseria meningitidis pili on bacterial interactions with three epithelial cell lines as well as human umbilical vein endothelial cells was studied using a panel of seven strains expressing Class I or Class II pili. Comparison of adherence of piliated and pilus-deficient variants of each strain to epithelial cells suggested that Class I pili may mediate bacterial adherence with all three epithelial cell lines. In contrast, Class II pili of the strains used did not increase bacterial adherence to Hep-2 larynx carcinoma cells, although an increase in adherence to Chang conjunctival and A549 lung carcinoma epithelial cells was observed in the Class II pili-expressing strains. In addition to these interclass functional variations, differences in adherence to epithelial cells were also observed among Class I and Class II strains. Functionally different pilin variants of one Class I strain, MC58, were obtained by single colony isolation. One piliated variant was identified which had concurrently lost the ability to adhere to both Chang and Hep-2 cells ('non-adherent' phenotype; adherence of less than 2 bacteria per cell). In addition, several adherent pilin variants were isolated from non-adherent Pil- and Pil+ bacteria by selection on Chang cells (adherence of 10-25 bacteria per cell). In contrast to epithelial cells, all variant pili, whether of Class I or Class II, adhered to endothelial cells in substantially larger numbers (greater than 50 bacteria per cell) and therefore implied the existence of distinct mechanisms in pilus-facilitated interactions of N. meningitidis with endothelial and epithelial cells.


Chromatographic Separation of the Codonocarpine Type Alkaloids from the Root Bark of Capparis decidua.
Various parts of the caper tree Capparis decidua have found application in traditional medicine. The isolation and structural elucidation of the codonocarpine type alkaloids contained in the root bark, however, is not trivial and has probably led to misinterpretation in the past. This protocol describes the extraction and chromatographic separation of the four major alkaloids of the root bark of Capparis decidua. The delivered samples of cadabicine, codonocarpine, isocodonocarpine and capparidisinine were suitable for their unambiguous structural elucidation by NMR, MS and MS/MS.


Effects of antenatal and postnatal environments on CD4 T-cell responses to Mycobacterium bovis BCG in healthy infants in the Gambia.
The Mycobacterium bovis BCG vaccine has a poor record of efficacy in low-income tropical settings. Against this background, we evaluated the immune response of infants to mycobacterial antigens over the 2 years following BCG vaccination at birth by measuring the gamma interferon (IFN-gamma), interleukin-2 (IL-2), and CD154 responses of CD4 T cells. Similar numbers of cells expressed IFN-gamma in infants, 4- to 5-year-old children, and adults, while CD154 was not expressed at comparable levels until the second year of infancy. The IL-2 response remained relatively low in infants, children, and adults but correlated negatively with mother's body mass index and was highest among infants born to Mandinka mothers. Similarly, infants born in the wet season had a stronger CD154 response than those born in the dry season throughout the 2 years of the study. We conclude that the prenatal and perinatal environments have a lasting effect on the response of infants to the BCG vaccine.


Transport mechanism and pH regulation of the Na+/H+ antiporter NhaA from Escherichia coli: an electrophysiological study.
Using an electrophysiological assay the activity of NhaA was tested in a wide pH range from pH 5.0 to 9.5. Forward and reverse transport directions were investigated at zero membrane potential using preparations with inside-out and right side-out-oriented transporters with Na(+) or H(+) gradients as the driving force. Under symmetrical pH conditions with a Na(+) gradient for activation, both the wt and the pH-shifted G338S variant exhibit highly symmetrical transport activity with bell-shaped pH dependences, but the optimal pH was shifted 1.8 pH units to the acidic range in the variant. In both strains the pH dependence was associated with a systematic increase of the K(m) for Na(+) at acidic pH. Under symmetrical Na(+) concentration with a pH gradient for NhaA activation, an unexpected novel characteristic of the antiporter was revealed; rather than being down-regulated, it remained active even at pH as low as 5. These data allowed a transport mechanism to advance based on competing Na(+) and H(+) binding to a common transport site and a kinetic model to develop quantitatively explaining the experimental results. In support of these results, both alkaline pH and Na(+) induced the conformational change of NhaA associated with NhaA cation translocation as demonstrated here by trypsin digestion. Furthermore, Na(+) translocation was found to be associated with the displacement of a negative charge. In conclusion, the electrophysiological assay allows the revelation of the mechanism of NhaA antiport and sheds new light on the concept of NhaA pH regulation.


Prognosis for a mid-infrared magnetic rotation spectrometer for the in situ detection of atmospheric free radicals.
Mid-infrared magnetic rotation spectroscopy (MRS) experiments on nitric oxide (NO) are quantitatively modeled by theoretical calculations. The verified theory is used to specify an instrument that can make in situ measurements on NO and NO(2) in the Earth's atmosphere at a sensitivity level of a few parts in 10(12) by volume per second. The prototype instrument used in the experiments has an extrapolated detection limit for NO of 30 parts in 10(9) for a 1-s integration time over a 12-cm path length. The detection limit is an extrapolation of experimental results to a signal-to-noise ratio of one, where the noise is considered to be one-half the peak-to-peak baseline noise. Also discussed are the various factors that can limit the sensitivity of a MRS spectrometer that uses liquid-nitrogen-cooled lead-salt diode lasers and photovoltaic detectors.


Prehospital triple antiplatelet therapy in patients with acute ST elevation myocardial infarction leads to better platelet aggregation inhibition and clinical outcome than dual antiplatelet therapy.
To assess whether prehospital initiation of high-dose tirofiban in addition to high-dose clopidogrel results in more adequate inhibition of platelet aggregation (IPA) and better clinical outcome after primary percutaneous coronary intervention (PCI). Prespecified two-centre substudy of the prospective, international, multicentre, placebo controlled Ongoing Tirofiban in Myocardial Infarction Evaluation trial 2 (On-TIME-2 trial). 648 of 964 (67%) patients in the On-TIME-2 trial with ST elevation myocardial infarction undergoing primary PCI were studied. Pre-PCI IPA after early prehospital initiation of high-bolus dose (25 μg/kg) tirofiban was compared to placebo in addition to acetylsalicylic acid, unfractionated heparin and 600 mg clopidogrel. IPA was measured at a median of 60 min after study medication administration. In all four tests: Fe induced platelet aggregation, ADP induced platelet aggregation, platelet function analyser (PFA)-100 (collagen-epinephrine and collagen-ADP cartridge) IPA was higher in patients pretreated with high-dose tirofiban (p<0.001 for all tests), even after >74 min of pretreatment. Patients in the highest quartile of IPA had less residual ST segment deviation 1 h post-PCI (p value for trend: p=0.001, 0.004, 0.001, 0.002 respectively). There was a significant relationship between PFA-100 (both cartridges) and major adverse cardiovascular events (MACE, p=0.028, p=0.035) and early thrombosis (p=0.009, p=0.007). 60 min of prehospital initiated antiplatelet treatment including high-dose tirofiban resulted in higher levels of IPA compared to pretreatment with acetylsalicylic acid and high-dose clopidogrel alone, even after longer pretreatment times. Levels of IPA were significantly related to ST resolution and MACE, including stent thrombosis. This substudy confirms the main findings of the On-TIME2 trial that clopidogrel alone is suboptimal, even at high dose and administered well in advance of primary PCI.


Toward cardiovascular MRI at 7 T: clinical needs, technical solutions and research promises.
To consider potential clinical needs, technical solutions and research promises of ultrahigh-field strength cardiovascular MR (CMR). A literature review is given, surveying advantages and disadvantages of CMR at ultrahigh fields (UHF). Key concepts, emerging technologies, practical considerations and applications of UHF CMR are provided. Examples of UHF CMR imaging strategies and their added value are demonstrated, including the numerous unsolved problems. A concluding section explores future directions in UHF CMR. UHF CMR can be regarded as one of the most challenging MRI applications. Image quality achievable at UHF is not always exclusively defined by signal-to-noise considerations. Some of the inherent advantages of UHF MRI are offset by practical challenges. But UHF CMR can boast advantages over its kindred lower field counterparts by trading the traits of high magnetic fields for increased temporal and/or spatial resolution. CMR at ultrahigh-field strengths is a powerful motivator, since speed and signal may be invested to overcome the fundamental constraints that continue to hamper traditional CMR. If practical challenges can be overcome, UHF CMR will help to open the door to new approaches for basic science and clinical research.


Fossil spores, pollen, and fishes from connecticut indicate early jurassic age for part of the newark group.
Palynologically productive localities have been found in the United States throughout the Newark Group basins, most of which had previously been assumed to be barren. Rich palynoflorules dominated by coniferous pollen of Circulina-Classopollis type, and well-preserved fossil fishes, including possible new semionotids, have been found in the Hartford basin. Palynological data indicate that the Newark Group has considerable time-stratigraphic range: Upper Triassic for the Cumnock Formation (North Carolina), the Vinita Beds (Virginia), and the upper New Oxford Formation (Pennsylvania), Rhaeto-Liassic for the Brunswick Formation (New Jersey), Portland Formation (Connecticut and Massachusetts), and the Shuttle Meadow Formation (Connecticut).


Inactivation of the spirochete recA gene results in a mutant with low viability and irregular nucleoid morphology.
Recently, we have shown the first evidence for allelic exchange in Leptospira spp. By using the same methodology, the cloned recA of Leptospira biflexa was interrupted by a kanamycin resistance cassette, and the mutated allele was then introduced into the L. biflexa chromosome by homologous recombination. The recA double-crossover mutant showed poor growth in liquid media and was considerably more sensitive to DNA-damaging agents such as mitomycin C and UV light than the wild-type strain. The efficiency of plating of the recA mutant was about 10% of that of the parent strain. In addition, microscopic observation of the L. biflexa recA mutant showed cells that are more elongated than those of the wild-type strain. Fluorescent microscopy of stained cells of the L. biflexa wild-type strain revealed that chromosomal DNA is distributed throughout most of the length of the cell. In contrast, the recA mutant showed aberrant nucleoid morphologies, i.e., DNA is condensed at the midcell. Our data indicate that L. biflexa RecA plays a major role in ensuring cell viability via mechanisms such as DNA repair and, indirectly, active chromosome partitioning.


Mitochondrial Redox Dysfunction and Environmental Exposures.
Mitochondria are structurally and biochemically diverse, even within a single type of cell. Protein complexes localized to the inner mitochondrial membrane synthesize ATP by coupling electron transport and oxidative phosphorylation. The organelles produce reactive oxygen species (ROS) from mitochondrial oxygen and ROS can, in turn, alter the function and expression of proteins used for aerobic respiration by post-translational and transcriptional regulation. New interest is emerging not only into the roles of mitochondria in disease development and progression but also as a target for environmental toxicants. Dysregulation of respiration has been linked to cell death and is a major contributor to acute neuronal trauma, peripheral diseases, as well as chronic neurodegenerative diseases, such as Parkinson's disease and Alzheimer's disease. Here, we discuss the mechanisms underlying the sensitivity of the mitochondrial respiratory complexes to redox modulation, as well as examine the effects of environmental contaminants that have well-characterized mitochondrial toxicity. The contaminants discussed in this review are some of the most prevalent and potent environmental contaminants that have been linked to neurological dysfunction, altered cellular respiration, and oxidation.


Statistical Verification of Anomaly in Chiral Angle Distribution of Air-Suspended Carbon Nanotubes.
Single-walled carbon nanotubes (SWCNT) have long attracted attention due to their distinct physical properties, depending on their chiral structures (chiralities). Clarifying their growth mechanism is important toward perfect chirality-controlled bulk synthesis. Although a correlation between the chirality distribution and the carbon atom configuration at an open tube edge has been predicted theoretically, lack of sufficient statistical data on metallic and semiconducting SWCNTs prohibited its verification. Here, we report statistical verification of the chirality distribution of 413 as-grown individual air-suspended SWCNTs with a length of over 20 μm using broadband Rayleigh spectroscopy. After excluding the impact of the difference in the number of possible SWCNT structures per chiral angle interval, the abundance profile with chiral angle exhibits an increasing trend with a distinct anomaly at a chiral angle of approximately 20°. These results are well explained considering the growth rate depending on armchair-shaped site configurations at the catalyst-nanotube interface.


Diagnostic accuracy of the Ottawa Ankle and Midfoot Rules: a systematic review with meta-analysis.
To review the diagnostic accuracy of the Ottawa Ankle and Midfoot Rules and explore if clinical features and/or methodological quality of the study influence diagnostic accuracy estimates. Systematic review with meta-analysis. MEDLINE, EMBASE, CINAHL, SPORTDiscus and Cochrane Library. Primary diagnostic studies reporting the accuracy of the Rules in people with ankle and/or midfoot injury were retrieved. Diagnostic accuracy estimates, overall and for subgroups (patient's age, profession of the assessor and setting of application), were made. Sensitivity analyses included studies with a low risk of bias and studies where all patients received radiographs. 66 studies were included. Ankle and Midfoot Rules presented similar accuracies, which were homogeneous and high for sensitivity and negative likelihood ratios and poor and heterogeneous for specificity and positive likelihood ratios (mean, 95% CI pooled sensitivity of Ankle Rules: 99.4%, 97.9% to 99.8%; specificity: 35.3%, 28.8% to 42.3%). Sensitivity of the Ankle Rules was higher in adults than in children, but the profession of the assessor did not appear to influence accuracy. Specificity was higher for Midfoot than for Ankle Rules. There were not enough studies to allow comparison according to setting of application. Studies with a low risk of bias and where all patients received radiographs provided lower accuracy estimates. Specificity heterogeneity was not explained by assessor training, use of imaging in all patients and low risk of bias. Study features and the methodological quality influence estimates of the diagnostic accuracy of the Ottawa Ankle and Midfoot Rules.


Immune derived opioidergic inhibition of viscerosensory afferents is decreased in Irritable Bowel Syndrome patients.
Alterations in the neuro-immune axis contribute toward viscerosensory nerve sensitivity and symptoms in Irritable Bowel Syndrome (IBS). Inhibitory factors secreted from immune cells inhibit colo-rectal afferents in health, and loss of this inhibition may lead to hypersensitivity and symptoms. We aimed to determine the immune cell type(s) responsible for opioid secretion in humans and whether this is altered in patients with IBS. The β-endorphin content of specific immune cell lineages in peripheral blood and colonic mucosal biopsies were compared between healthy subjects (HS) and IBS patients. Peripheral blood mononuclear cell (PBMC) supernatants from HS and IBS patients were applied to colo-rectal sensory afferent endings in mice with post-inflammatory chronic visceral hypersensitivity (CVH). β-Endorphin was identified predominantly in monocyte/macrophages relative to T or B cells in human PBMC and colonic lamina propria. Monocyte derived β-endorphin levels and colonic macrophage numbers were lower in IBS patients than healthy subjects. PBMC supernatants from healthy subjects had greater inhibitory effects on colo-rectal afferent mechanosensitivity than those from IBS patients. The inhibitory effects of PBMC supernatants were more prominent in CVH mice compared to healthy mice due to an increase in μ-opioid receptor expression in dorsal root ganglia neurons in CVH mice. Monocyte/macrophages are the predominant immune cell type responsible for β-endorphin secretion in humans. IBS patients have lower monocyte derived β-endorphin levels than healthy subjects, causing less inhibition of colonic afferent endings. Consequently, altered immune function contributes toward visceral hypersensitivity in IBS.


Triple therapies plus different probiotics for Helicobacter pylori eradication.
The Helicobacter pylori (H. pylori) cure rate following standard triple therapies is decreasing worldwide. Therefore, further approaches aimed to improve standard triple therapy efficacy should be attempted. This prospective, pilot study aimed to evaluate the therapeutic role of either Lactobacillus reuteri (L. reuteri) or a high concentration of probiotics in addition to standard triple therapies for H. pylori eradication. The study enrolled 65 consecutive dyspeptic patients with H. pylori infection. All patients underwent upper endoscopy with gastric biopsies. Patients were assigned to receive one of the following therapies: (a) standard 7-day triple; (b) the same 7-day triple therapy plus L. reuteri supplementation; (c) the same 7-day triple therapy plus a probiotic mixture; and d) a 14-day standard triple therapy plus a probiotic mixture. H. pylori eradication was checked by using a 13C-urea breath test performed 4-6 weeks after treatment. No therapy regimen achieved > 80% eradication rate at both intention-to-treat (ITT) and per protocol (PP) analyses. Although the 14-day therapy plus a probiotic mixture tended to achieve higher eradication rate (71%), no statistically significant difference emerged among the different therapy regimens tested (range: 53-71%). The lowest incidence of side-effects was observed following the 7-day therapy plus L. reuteri (6%) and highest with the 14-day triple therapy plus probiotic mixture (33%), although the difference failed to reach the statistically significance. In conclusion, our data found that 7-14 days triple therapy with or without probiotic supplementation failed to achieved acceptable H. pylori eradication rates.


Combining refuges with transgenic insect releases for the management of an insect pest with non-recessive resistance to Bt crops in agricultural landscapes.
Reinforcing the high-dose/refuge strategy with releases of transgenic insects has been suggested as a method for simultaneously managing agricultural pest populations and resistance to transgenic crops. Theoretical and empirical studies have shown that these approaches can work when deployed against closed populations and the assumptions of the HDR strategy are met. However, field-evolved resistance is often linked to non-recessive resistance or refuge non-compliance, and pest management regimes are likely to take place at the landscape-level. It is therefore important to understand how effective such strategies are when resistance is non-recessive, and how they could be employed in agricultural landscapes. We developed a spatially-explicit model to investigate the efficacy of strategies combining refuges with transgenic insect releases to manage a pest with non-recessive resistance in agricultural landscapes. We compared two release strategies, area-wide releases and localised releases targeted at population hotspots, and analysed the effects of refuge and release parameters on population and resistance dynamics. Area-wide releases reliably achieved landscape-level pest eradication. Localised releases also eradicated the pest when low release thresholds were combined with high release ratios, and maintained the pest at low densities when insufficient to achieve extinction. Reinforcing refuges with localised releases also greatly enhanced the probability of resistance extinction. However, when resistance remained in the population, localised releases prevented resistance from reaching fixation rather than greatly delaying or reversing resistance evolution. Our work indicates that combining refuges with simple release policies is effective for landscape-level pest suppression when the HDR assumptions are violated, but more nuanced release strategies may be required to enhance the benefits to resistance management.


Productivity outcomes for recent grants and fellowships awarded by the American Osteopathic Association Bureau of Research.
The objective of the present study was to evaluate productivity outcome measures for recent research grants and fellowships awarded through the American Osteopathic Association (AOA) Bureau of Research. Recipients of grants and fellowships that were awarded between 1995 and 2001 were contacted by mail, e-mail, or telephone and asked to provide information about publications, resulting grant awards, advances in clinical care, or other notable products that were generated from their projects. For grants funded between 1995 and 1998, 76% of principal investigators reported a notable product from their study. By contrast, for grants funded between 1999 and 2001, only 31% reported a notable outcome. This difference most likely can be attributed to the lag time between the awarding of a grant and actual completion of the project, the processing of the data, and the publication of the results. Several recipients of 1999-2001 grants were optimistic about eventually generating a notable product. Most (79%) of the 1995-2001 fellows met the requirements for successful completion of their project. Many fellows exceeded the minimal requirement by publishing their results, continuing research activity, attracting extramural grant monies, or entering an academic position. It appeared that a much larger proportion of osteopathic fellows went on to academic careers than their counterparts who did not have fellowship training. From 1995 to 2001, the AOA Bureau of Research awarded dollars 3,072,140 in research grants and fellowships. To date, these awards have helped the recipients bring an additional dollars 5,659,329 of extramural funds for research at osteopathic institutions. The Bureau of Research grant and fellowship programs have been successful both scientifically and in terms of financial outcomes.


Web monitoring of emerging animal infectious diseases integrated in the French Animal Health Epidemic Intelligence System.
Since 2013, the French Animal Health Epidemic Intelligence System (in French: Veille Sanitaire Internationale, VSI) has been monitoring signals of the emergence of new and exotic animal infectious diseases worldwide. Once detected, the VSI team verifies the signals and issues early warning reports to French animal health authorities when potential threats to France are detected. To improve detection of signals from online news sources, we designed the Platform for Automated extraction of Disease Information from the web (PADI-web). PADI-web automatically collects, processes and extracts English-language epidemiological information from Google News. The core component of PADI-web is a combined information extraction (IE) method founded on rule-based systems and data mining techniques. The IE approach allows extraction of key information on diseases, locations, dates, hosts and the number of cases mentioned in the news. We evaluated the combined method for IE on a dataset of 352 disease-related news reports mentioning the diseases involved, locations, dates, hosts and the number of cases. The combined method for IE accurately identified (F-score) 95% of the diseases and hosts, respectively, 85% of the number of cases, 83% of dates and 80% of locations from the disease-related news. We assessed the sensitivity of PADI-web to detect primary outbreaks of four emerging animal infectious diseases notifiable to the World Organisation for Animal Health (OIE). From January to June 2016, PADI-web detected signals for 64% of all primary outbreaks of African swine fever, 53% of avian influenza, 25% of bluetongue and 19% of foot-and-mouth disease. PADI-web timely detected primary outbreaks of avian influenza and foot-and-mouth disease in Asia, i.e. they were detected 8 and 3 days before immediate notification to OIE, respectively.


Predicting risk of osteoporotic and hip fracture in the United Kingdom: prospective independent and external validation of QFractureScores.
To evaluate the performance of the QFractureScores for predicting the 10 year risk of osteoporotic and hip fractures in an independent UK cohort of patients from general practice records. Prospective cohort study. 364 UK general practices contributing to The Health Improvement Network (THIN) database. 2.2 million adults registered with a general practice between 27 June 1994 and 30 June 2008, aged 30-85 (13 million person years), with 25,208 osteoporotic fractures and 12,188 hip fractures. First (incident) diagnosis of osteoporotic fracture (vertebra, distal radius, or hip) and incident hip fracture recorded in general practice records. Results from this independent and external validation of QFractureScores indicated good performance data for both osteoporotic and hip fracture end points. Discrimination and calibration statistics were comparable to those reported in the internal validation of QFractureScores. The hip fracture score had better performance data for both women and men. It explained 63% of the variation in women and 60% of the variation in men, with areas under the receiver operating characteristic curve of 0.89 and 0.86, respectively. The risk score for osteoporotic fracture explained 49% of the variation in women and 38% of the variation in men, with corresponding areas under the receiver operating characteristic curve of 0.82 and 0.74. QFractureScores were well calibrated, with predicted risks closely matching those across all 10ths of risk and for all age groups. QFractureScores are useful tools for predicting the 10 year risk of osteoporotic and hip fractures in patients in the United Kingdom.


Oxidation of xenobiotics by recombinant human cytochrome P450 1B1.
Human cytochrome P450 (P450) 1B1 (CYP1B1) has recently been shown to be an important enzyme in the activation of diverse procarcinogens such as arylarenes, nitroarenes, and arylamines to reactive metabolites that cause DNA damage in the cells. However, it is not known whether this P450 enzyme also plays roles in the oxidation of certain drugs or model substrates commonly used in P450 assays. We examined the substrate oxidation activities of recombinant human CYP1B1 in yeast microsomes and compared these activities with those catalyzed by reconstituted systems containing recombinant CYP1A1 and CYP1A2 which were isolated from membranes of Escherichia coli in which respective cDNAs have been expressed. Catalytic activities towards some of the model substrates of other human P450 enzymes including CYP2A6, 2C9, 2C19, 2D6, 2E1, and 3A4 were also determined and compared. CYP1B1 catalyzed benzo[a]pyrene 3-hydroxylation at rates lower than those of CYP1A1 but higher than those of CYP1A2. The activity towards 7-ethoxyresorufin O-deethylation catalyzed by CYP1B1 was about one-tenth of that of CYP1A1, but the Km values were lower for CYP1B1 than those for CYP1A1 and CYP1A2. CYP1B1 was also able to catalyze the oxidation of theophylline and caffeine, two prototypic substrates for CYP1A2. CYP1B1 did not oxidize other typical P450 substrates such as coumarin, tolbutamide, S-mephenytoin, chlorzoxazone, nifedipine, and testosterone, while low rates of oxidation of bufuralol and 7-ethoxycoumarin were found for CYP1B1. These results indicate that CYP1B1 has catalytic activities overlapping CYP1A1 and CYP1A2 with respect to the oxidation of drugs and model P450 substrates, although the relative catalytic roles in these three P450 enzymes differ depending upon the substrates examined. A distinct marker activity of CYP1B1 has not been identified.


Early differentiation of the Earth and the problem of mantle siderophile elements: a new approach.
The long-standing problem of the excess abundances of siderophile elements in the mantle can be resolved by considering an equilibrium core-mantle differentiation in the earth at 3000 to 3500 kelvin. This high-temperature differentiation results in mantle siderophile element abundances that closely match the observed values. Some lithophile (light) elements could enter the core in this process as is necessary to account for its low density. The abundances of siderophile elements in the mantle are consistent with the conclusion derived from the recent physical models that the earth was molten during accretion.


Exposure to non-therapeutic INR in a high risk cardiovascular patient: potential hazard reduction with genotype-guided warfarin (Coumadin) dosing.
A case to illustrate the utility of genetic screening in warfarin (Coumadin) management is reported. A 45 year-old woman of Puerto Rican ancestry was admitted to the emergency room twice within one month with chest pain. She was diagnosed with congestive heart failure, which was stabilized both times. At her second release, warfarin therapy was initiated at 5 mg/ day to prevent thrombus formation and was lowered to 3.75 mg/day at day 7 by her primary physician. International Normalized Ratio (INR) test results in the follow-up period at days 1, 7, and 10 of warfarin therapy were 4.5, 6.5, and 7.3, respectively-far in excess of the therapeutic range, despite the lower dosage in effect from day 7 onward. The patient achieved target INR over the next 43 days after downward adjustment of the dose to a dose of 1.5 mg/day by trial and error. DNA-typing specific for the CYP2C9*2,*3,*4,*5,*6 alleles and seven variants in the VKORC1 gene, including the VKORC1-1639 G > A polymorphism, revealed the presence of combinatorial CYP2C9*2/*3 and VKORC1-1639 G/A genotypes in this patient. Entering the patient's demographic and genotype status data into independent algorithms available in the public domain to predict effective warfarin dose yielded predicted doses which ranged from 1.5 to 1.8 mg/day. Notably, the prediction of 1.5 mg/day, which was generated by the online resource www.warfarindosing.org, coincided with the patient's actual effective warfarin dose. We conclude that the rapid rise in INR observed upon the initiation of warfarin therapy and the final effective warfarin dose of 1.5 mg/day, are attributable in some part to the presence of two minor alleles in CYP2C9, which together significantly reduce warfarin metabolism. Warfarin genotyping can therefore inform the clinician of the predicted effective warfarin dose. The results highlight the potential for warfarin genetic testing to improve patient care.


Prognostic impact of ATM mutations in patients with metastatic colorectal cancer.
Tumors bearing homologous recombination deficiency are extremely sensitive to DNA double strand breaks induced by several chemotherapeutic agents. ATM gene, encoding a protein involved in DNA damage response, is frequently mutated in colorectal cancer (CRC), but its potential role as predictive and prognostic biomarker has not been fully investigated. We carried out a multicenter effort aimed at defining the prognostic impact of ATM mutational status in metastatic CRC (mCRC) patients. Mutational profiles were obtained by means of next-generation sequencing. Overall, 35 out of 227 samples (15%) carried an ATM mutation. At a median follow-up of 56.6 months, patients with ATM mutated tumors showed a significantly longer median overall survival (OS) versus ATM wild-type ones (64.9 vs 34.8 months; HR, 0.50; 95% CI, 0.29-0.85; P = 0.01). In the multivariable model, ATM mutations confirmed the association with longer OS (HR, 0.57; 95% CI, 0.33-0.98; P = 0.04). The prognostic impact of ATM mutations was independent from TP53 mutational status and primary tumor location. High heterogeneity score for ATM mutations, possibly reflecting the loss of wild-type allele, was associated with excellent prognosis. In conclusion, we showed that ATM mutations are independently associated with longer OS in patients with mCRC.


Reliable Isolation of Central Nervous System Microvessels Across Five Vertebrate Groups.
Isolation of microvessels from the central nervous system (CNS) is commonly performed by combining cortical tissue from multiple animals, most often rodents. This approach limits the interrogation of blood-brain barrier (BBB) properties to the cortex and does not allow for individual comparison. This project focuses on the development of an isolation method that allows for the comparison of the neurovascular unit (NVU) from multiple CNS regions: cortex, cerebellum, optic lobe, hypothalamus, pituitary, brainstem, and spinal cord. Moreover, this protocol, originally developed for murine samples, was successfully adapted for use on CNS tissues from small and large vertebrate species from which we are also able to isolate microvessels from brain hemisphere white matter. This method, when paired with immunolabeling, allows for quantitation of protein expression and statistical comparison between individuals, tissue type, or treatment. We proved this applicability by evaluating changes in protein expression during experimental autoimmune encephalomyelitis (EAE), a murine model of a neuroinflammatory disease, multiple sclerosis. Additionally, microvessels isolated by this method could be used for downstream applications like qPCR, RNA-seq, and Western blot, among others. Even though this is not the first attempt to isolate CNS microvessels without the use of ultracentrifugation or enzymatic dissociation, it is unique in its adeptness for the comparison of single individuals and multiple CNS regions. Therefore, it allows for investigation of a range of differences that may otherwise remain obscure: CNS portions (cortex, cerebellum, optic lobe, brainstem, hypothalamus, pituitary, and spinal cord), CNS tissue type (gray or white matter), individuals, experimental treatment groups, and species.


A quantitative mechanistic PK/PD model directly connects Btk target engagement and in vivo efficacy.
Correlating target engagement with in vivo drug activity remains a central challenge in efforts to improve the efficiency of drug discovery. Previously we described a mechanistic pharmacokinetic-pharmacodynamic (PK/PD) model that used drug-target binding kinetics to successfully predict the in vivo efficacy of antibacterial compounds in models of Pseudomonas aeruginosa and Staphylococcus aureus infection. In the present work we extend this model to quantitatively correlate the engagement of Bruton's tyrosine kinase (Btk) by the covalent inhibitor CC-292 with the ability of this compound to reduce ankle swelling in an animal model of arthritis. The modeling studies include the rate of Btk turnover and reveal the vulnerability of Btk to engagement by CC-292.


Differential Effects of Encoding Instructions on Brain Activity Patterns of Item and Associative Memory.
Evidence from neuroimaging studies suggests a critical role of hippocampus and inferior frontal gyrus (IFG) in associative relative to item encoding. Here, we investigated similarities and differences in functional brain correlates for associative and item memory as a function of encoding instruction. Participants received either incidental (animacy judgments) or intentional encoding instructions while fMRI was employed during the encoding of associations and items. In a subsequent recognition task, memory performance of participants receiving intentional encoding instructions was higher compared with those receiving incidental encoding instructions. Furthermore, participants remembered more items than associations, regardless of encoding instruction. Greater brain activation in the left anterior hippocampus was observed for intentionally compared with incidentally encoded associations, although activity in this region was not modulated by the type of instruction for encoded items. Furthermore, greater activity in the left anterior hippocampus and left IFG was observed during intentional associative compared with item encoding. The same regions were related to subsequent memory of intentionally encoded associations and were thus task relevant. Similarly, connectivity of the anterior hippocampus to the right superior temporal lobe and IFG was uniquely linked to subsequent memory of intentionally encoded associations. Our study demonstrates the differential involvement of anterior hippocampus in intentional relative to incidental associative encoding. This finding likely reflects that the intent to remember triggers a specific binding process accomplished by this region.


[Assessment of accuracy of parents' perception of their 4-36 months old children's picky eating behavior].
To evaluate the accuracy of parents' perception of whether their child is a picky eater and the specific food category the children avoideating according to the food frequency questionnaire. This research selected 1 663 infants aged 4-36 months receiving non-diary complimentary food from maternal, infants, nutrition and growth study (MING Study) in 8 Chinese cities in which a combination of systematic cluster random sampling and purposive sampling was used. The general information, dietary status and picky eating status were collected through a self-designed questionnaire from the caregiver of the children. According to the parents' perception, the children were classified into picky/non-picky groups or avoid/non-avoid to a specific food category groups. Mann-Whitney U test was used to compare between the groups. The reported percentage of picky eaters increased from 7.37% in 4-6 months old infants to 36.20% in 25-36 months old infants. Most picky infants in 4-6 months and 7-12 months old infants avoided eating dairy food (25% and 24%); while most picky toddlers aged 13-24 months and 25-36 months avoided eating vegetables (26.92% and 47.46%). The infants aged 4-6 months and 7-12 months who were perceived as picky by their parents took more kinds of food (8 and 19.5 kinds) than those who were not (6 and 18 kinds), while the picky toddlers aged 13-24 months and 25-36 months took fewer kinds of food (28.5 and 34 kinds for picky eaters, 31 and 37 kinds for non-picky eaters). The parents of infants aged 4-6 months judged correctly in every category of food without any statistical significance; the parents of 7-12 months old infants judged correctly only in dairy food and eggs with statistical significance; those of 13-24 months old infants judged correctly in every food category except for vegetables with statistically significant difference in the category of eggs; those of 25-36 months old toddlers misjudged in dairy, beans and grains with no statistically significant difference in every category. Parents tend to misjudge their children's picky eating behavior before the first 12 months of the child, and tend to make a more accurate perception after the 12th month.


Immunochemical characterization on pathological oligomers of mutant Cu/Zn-superoxide dismutase in amyotrophic lateral sclerosis.
Dominant mutations in Cu/Zn-superoxide dismutase (SOD1) gene cause a familial form of amyotrophic lateral sclerosis (SOD1-ALS) with accumulation of misfolded SOD1 proteins as intracellular inclusions in spinal motor neurons. Oligomerization of SOD1 via abnormal disulfide crosslinks has been proposed as one of the misfolding pathways occurring in mutant SOD1; however, the pathological relevance of such oligomerization in the SOD1-ALS cases still remains obscure. We prepared antibodies exclusively recognizing the SOD1 oligomers cross-linked via disulfide bonds in vitro. By using those antibodies, immunohistochemical examination and ELISA were mainly performed on the tissue samples of transgenic mice expressing mutant SOD1 proteins and also of human SOD1-ALS cases. We showed the recognition specificity of our antibodies exclusively toward the disulfide-crosslinked SOD1 oligomers by ELISA using various forms of purified SOD1 proteins in conformationally distinct states in vitro. Furthermore, the epitope of those antibodies was buried and inaccessible in the natively folded structure of SOD1. The antibodies were then found to specifically detect the pathological SOD1 species in the spinal motor neurons of the SOD1-ALS patients as well as the transgenic model mice. Our findings here suggest that the SOD1 oligomerization through the disulfide-crosslinking associates with exposure of the SOD1 structural interior and is a pathological process occurring in the SOD1-ALS cases.


The emergency medicine evaluation and management of the patient with cirrhosis.
Cirrhosis is a significant cause of death in the U.S. and has a variety of causes, most commonly Hepatitis C and alcohol. Liver fibrosis and nodule formation result in significant complications due to portal system hypertension. There are several deadly complications emergency physicians must consider. Provide an evidence-based update for the resuscitation of decompensating cirrhotic patients and an overview of cirrhosis complications. Cirrhosis is a common condition in the U.S. and leads to several deadly complications. The disease develops from liver fibrosis, elevating portal pressures and modifying patient hemodynamics. Cirrhosis results in significant anatomic and physiologic modifications involving the gastrointestinal, cardiopulmonary, neurologic, renal, immunologic, and hematologic systems. The disease can be divided into compensated and decompensated states, with decompensation associated with significant morbidity and mortality. Complications include variceal hemorrhage, ascites, increased risk of bacterial infection, spontaneous bacterial peritonitis (SBP), hepatic encephalopathy, hepatorenal syndrome, hepatopulmonary syndrome, umbilical hernia, and hepatic hydrothorax. Resuscitation including airway and circulation measures is paramount in these patients, and several new techniques are offered for the approach to intubation and resuscitation for patients with severe cirrhosis. Decompensating cirrhotics may require extensive resuscitation, and knowledge of the evaluation and management of complications associated with cirrhosis can improve care for patients with severe liver disease.


Modeling and deadlock avoidance of automated manufacturing systems with multiple automated guided vehicles.
An automated manufacturing system (AMS) contains a number of versatile machines (or workstations), buffers, an automated material handling system (MHS), and is computer-controlled. An effective and flexible alternative for implementing MHS is to use automated guided vehicle (AGV) system. The deadlock issue in AMS is very important in its operation and has extensively been studied. The deadlock problems were separately treated for parts in production and transportation and many techniques were developed for each problem. However, such treatment does not take the advantage of the flexibility offered by multiple AGVs. In general, it is intractable to obtain maximally permissive control policy for either problem. Instead, this paper investigates these two problems in an integrated way. First we model an AGV system and part processing processes by resource-oriented Petri nets, respectively. Then the two models are integrated by using macro transitions. Based on the combined model, a novel control policy for deadlock avoidance is proposed. It is shown to be maximally permissive with computational complexity of O (n2) where n is the number of machines in AMS if the complexity for controlling the part transportation by AGVs is not considered. Thus, the complexity of deadlock avoidance for the whole system is bounded by the complexity in controlling the AGV system. An illustrative example shows its application and power.


Perspectives on the state of insect transgenics.
Genetic transformation is a critical component to the fundamental genetic analysis of insect species and holds great promise for establishing strains that improve population control and behavior for practical application. This is especially so for insects that are disease vectors, many of which are currently subject to genomic sequence analysis, and intensive population control measures that must be improved for better efficacy and cost-effectiveness. Transposon-mediated germ-line transformation has been the ultimate goal for most fundamental and practical studies, and impressive strides have been made in recent development of transgene vector and marker systems for several mosquito species. This has resulted in rapid advances in functional genomic sequence analysis and new strategies for biological control based on conditional lethality. Importantly, advances have also been made in our ability to use these systems more effectively in terms of enhanced stability and targeting to specific genomic loci. Nevertheless, not all insects are currently amenable to germ-line transformation techniques, and thus advances in transient somatic expression and paratransgenesis have also been critical, if not preferable for some applications. Of particular importance is how this technology will be used for practical application. Early ideas for population replacement of indigenous pests with innocuous transgenic siblings by transposon-vector spread, may require reevaluation in terms of our current knowledge of the behavior of transposons currently available for transformation. The effective implementation of any control program using released transgenics, will also benefit from broadening the perspective of these control measures as being more mainstream than exotic.


Synergistic Antitumor Effects of Berbamine and Paclitaxel through ROS/Akt Pathway in Glioma Cells.
In our preliminary study, Berbamine (BA), one of the most commonly used traditional Chinese medicines, was effective in inducing the intracellular ROS levels. Since the regulation of cellular antioxidant capacity is crucial to the sensitivity of Ptx, it is feasible to propose that sensitizing cells to Ptx can be achieved through increasing the antioxidant capacity by codelivering BA. Cytotoxicity test demonstrates that either single or combinational treatment of BA and Ptx dose-dependently inhibits the proliferation of U-87 cells. Median-effect analysis clearly proves the synergistic anticancer effect between BA and Ptx. Combinational treatment of both drugs induced more intracellular ROS generation than either of the drugs did. Cotreatment of NAC could partially reverse the ROS generation and ameliorate the cytotoxicity induced by BA plus Ptx. Moreover, sequential activation of ROS-dependent phosphor-Akt expression was dose-dependently inhibited by the combinational application of BA and Ptx, which was more significantly effective than the single treatment of either BA or Ptx. Additionally, the coadministration of BA and Ptx shows the strongest tumor delaying effect in a U87 xenograft model, demonstrating the synergism between two drugs. Therefore, BA is a promising adjuvant to traditional chemotherapy, especially in combination with Ptx, to treat malignant glioma.


Environmental optimization of chromium recovery from tannery sludge using a life cycle assessment approach.
Life cycle assessment (LCA) was used to evaluate the environmental impact of an oxidative chromium recovery method from tannery sludge, in comparison with the usual landfilling process. Three improvement options (water reduction, byproduct use and anaerobic sludge digestion) were considered. The results showed that the proposed chromium recovery process would be better environmentally than conventional landfilling in all the evaluated impact categories if the amount of chromium recovered was 43 kg per ton of sludge. This amount could be recovered if the chromium concentration was about 20 times higher than that considered in this study. Alternatively, a lower chromium concentration would produce a better result if the recovery method was optimized and implemented at industrial rather than laboratory scale, and if more accurate data were provided on environmental credits for avoiding the chromium production process. Thus, the recovery method is environmentally beneficial when tannery sludge contains a chromium concentration of about 100,000 ppm. According to the literature, such concentrations are not unusual. The results could serve as the basis for further environmental improvements in chromium recovery and tannery sludge management and should be used in decision-making processes, especially for end-of-pipe treatments.


The effect of 3-aminobenzamide, inhibitor of poly(ADP-ribose) polymerase, on human osteosarcoma cells.
This study demonstrates that in human osteosarcoma cells treatment with 3-aminobenzamide (3-AB), a potent inhibitor of poly(ADP-ribose) polymerase (PARP), induces morphological and biochemical features of differentiation, the duration of which depends on whether or not the normal RB gene is expressed. In Saos-2 cells expressing a non-functional Rb protein, 3-AB treatment induced the formation of transient, short dendritic-like protrusions. In RB-transfected-Saos-2 cells (a clone previously generated in our laboratory that shows stable expression of wild-type Rb protein), 3-AB induced marked and prolonged changes with the formation of long dendritic-like protrusions and the appearance of stellate (osteocyte-like) cells. In MG-63 cells producing a wild-type Rb protein, 3-AB treatment had more marked effects, with a larger number of cells assuming the stellate appearance of osteocytes, which were connected to each other via junctions resembling small channels. Regardless of cell type, at some point after 3-AB treatment the differentiative attempt failed and the cells died. Death was apoptotic, as demonstrated by chromatin condensation and fragmentation, specific cleavage of PARP and Lamin-B, processing of caspase-3 and the appearance of Bax immunoreactive species. Enzymatic assay and RT-PCR of alkaline phosphatase (ALP) - an enzyme whose levels markedly decrease when osteoblasts undergo terminal differentiation into osteocytes - showed that 3-AB treatment markedly lowered ALP expression. Simultaneously, 3-AB treatment markedly increased the expression of CD44, a transmembrane multifunctional adhesion molecule and sensitive marker of osteocytic differentiation. This study hypothesizes a cross-talk between pRb and PARP and suggests that PARP may be a useful target for anticancer drugs.