Pulmonary clearance of encapsulated and unencapsulated Haemophilus influenzae strains.
A mouse model system was employed to investigate the temporal pattern of pulmonary clearance of Haemophilus influenzae and to evaluate the effect of the type b polysaccharide capsule on this clearance pattern. The lungs of BALB/c mice were inoculated with boluses of several different H. influenzae strains via an endobronchial catheter. A fully encapsulated H. influenzae type b strain multiplied readily in the lungs for at least 6 h and then was eventually cleared from the lungs over the next 18 h. The pulmonary clearance pattern obtained with an unencapsulated variant of this H. influenzae type b strain was identical to that obtained with the fully encapsulated parent strain. Two nontypable H. influenzae strains isolated by transtracheal aspiration of patients with acute H. influenzae pneumonia also multiplied in the lung and resisted significant clearance for at least 6 h after inoculation. Bolus deposition of either H. influenzae type b or nontypable H. influenzae in the lungs resulted in an eventual influx of polymorphonuclear leukocytes into the alveoli. The observed delay in clearance of all these strains suggests that resident host defense mechanisms must be augmented for clearance to occur. Furthermore, these data indicate that one or more factors other than the Haemophilus capsule are important bacterial determinants of pulmonary clearance of H. influenzae.


UBC-9 Acts in GABA Neurons to Control Neuromuscular Signaling in C. elegans.
Regulation of excitatory to inhibitory signaling balance is essential to nervous system health and is maintained by numerous enzyme systems that modulate the activity, localization, and abundance of synaptic proteins. SUMOylation is a key post-translational regulator of protein function in diverse cells, including neurons. There, its role in regulating synaptic transmission through pre- and postsynaptic effects has been shown primarily at glutamatergic central nervous system synapses, where the sole SUMO-conjugating enzyme Ubc9 is a critical player. However, whether Ubc9 functions globally at other synapses, including inhibitory synapses, has not been explored. Here, we investigated the role of UBC-9 and the SUMOylation pathway in controlling the balance of excitatory cholinergic and inhibitory GABAergic signaling required for muscle contraction in Caenorhabditis elegans. We found inhibition or overexpression of UBC-9 in neurons modestly increased muscle excitation. Similar and even stronger phenotypes were seen with UBC-9 overexpression specifically in GABAergic neurons, but not in cholinergic neurons. These effects correlated with accumulation of synaptic vesicle-associated proteins at GABAergic presynapses, where UBC-9 and the C. elegans SUMO ortholog SMO-1 localized, and with defects in GABA-dependent behaviors. Experiments involving expression of catalytically inactive UBC-9 [UBC-9(C93S)], as well as co-expression of UBC-9 and SMO-1, suggested wild type UBC-9 overexpressed alone may act via substrate sequestration in the absence of sufficient free SUMO, underscoring the importance of tightly regulated SUMO enzyme function. Similar effects on muscle excitation, GABAergic signaling, and synaptic vesicle localization occurred with overexpression of the SUMO activating enzyme subunit AOS-1. Together, these data support a model in which UBC-9 and the SUMOylation system act at presynaptic sites in inhibitory motor neurons to control synaptic signaling balance in C. elegans. Future studies will be important to define UBC-9 targets at this synapse, as well as mechanisms by which UBC-9 and the SUMO pathway are regulated.


Controlling the growth rate of cancer cell.
To control the rate of growth of the cancer cell is the objective of this paper. In cancer, the rate of the growth of the cancer cell is indefinite. This paper proposes a method to transform into definite rate of growth of the cancer cell from indefinite. This indefiniteness lies with the set of unknown elements. This paper finds these unknown elements by matrix method.


Suppression of intra-articular responses to interleukin-1 by transfer of the interleukin-1 receptor antagonist gene to synovium.
We have developed an ex vivo method for delivering genes to the synovial lining of joints and expressing them intra-articularly. The present studies were designed to determine whether transfer of a human interleukin-1 receptor antagonist protein (IRAP) gene by this method was able to antagonize the intra-articular actions of interleukin-1. Intra-articular injections of human recombinant interleukin-1 beta (hrIL-1 beta) into the knees of control rabbits provoked a marked leukocytic infiltrate into the joint space, severe synovial thickening and hypercellularity, and loss of proteoglycans from articular cartilage. Genetically modified knees contained several nanograms of human IRAP and inhibited each of these effects of IL-1 beta. These data demonstrate for the first time that delivery of an appropriate gene to joints can prevent intra-articular pathology. Such findings permit cautious optimism about the eventual development of a gene treatment for arthritis and other disorders of the joint.


Effects of combination of intrathecal lidocaine and two doses of intrathecal midazolam on post-operative pain in patients undergoing herniorrhaphy: a randomized controlled trial.
Assessment of the effect of combination of intrathecal midazolam and lidocaine on postoperative pain was the aim of this study. This randomized controlled trial was performed during 2007 in a teaching hospital of Arak University of Medical Sciences. Forty five male patients who were candidates for elective inguinal herniorrhaphy entered the study and randomly divided into three groups of control (lidocaine 5% plus normal saline), M 0.5 (lidocaine 5% and midazolam 0.5 mg) and M 1.0 (lidocaine 5% and midazolam 1 mg) according intrathecal solution injected for spinal anesthesia. Mean arterial blood pressure, heart rate, post-operative pain, narcotic requirements and complications (nausea, vomiting, pruritic, headache, hypotension and bradycardia) were recorded. The severity of post-operative pain was lowest in M 1.0 group in all postoperative measurements except at 2 h after operation. With regard of complications, only there was significant difference in vomiting between three groups which had the highest frequency in M 0.5 group. No severe hypotension was seen; though, bradycardia occurred in one patient in M 0.5 group which needed treatment. Present findings suggest that administration of intrathecal midazolam (especially 1 mg) together with lidocaine is effective in reducing post-operative pain in patients undergoing open inguinal herniorrhaphy and is not associated with adverse effect.


Automatic bifurcation detection in coronary IVUS sequences.
In this paper, we present a fully automatic method which identifies every bifurcation in an intravascular ultrasound (IVUS) sequence, the corresponding frames, the angular orientation with respect to the IVUS acquisition, and the extension. This goal is reached using a two-level classification scheme: first, a classifier is applied to a set of textural features extracted from each image of a sequence. A comparison among three state-of-the-art discriminative classifiers (AdaBoost, random forest, and support vector machine) is performed to identify the most suitable method for the branching detection task. Second, the results are improved by exploiting contextual information using a multiscale stacked sequential learning scheme. The results are then successively refined using a-priori information about branching dimensions and geometry. The proposed approach provides a robust tool for the quick review of pullback sequences, facilitating the evaluation of the lesion at bifurcation sites. The proposed method reaches an F-Measure score of 86.35%, while the F-Measure scores for inter- and intraobserver variability are 71.63% and 76.18%, respectively. The obtained results are positive. Especially, considering the branching detection task is very challenging, due to high variability in bifurcation dimensions and appearance.


Experimental mouse muscle damage: the importance of external calcium.
The involvement of extracellular calcium in experimental muscle damage has been studied in an isolated mouse soleus muscle preparation. The enzyme efflux and ultrastructural damage seen after excessive contractile activity were markedly reduced when the extracellular calcium was withdrawn. Low extracellular calcium also protected against the large enzyme efflux seen after treatment with low concentrations of detergent. Treatment of the muscle with the calcium ionophore A 23187 caused significant release of enzyme from the muscle. Nifedipine did not prevent the enzyme release after stimulation and although in some circumstances verapamil appeared to have some protective effect this was probably due to a local anaesthetic action on the muscle and not to any specific effect on calcium movement. It is concluded that extracellular calcium is important in mediating at least the two forms of muscle damage studied here.


[Evolution of urinary lithiasis composition in a Spanish health care area over the last four decades].
To provide an updated epidemiological description of urinary lithiasis in a health area in the Western hemisphere over the past four decades. 2704 urinary lithiases analysed in our institution between 1980 and 2015 were retrospectively reviewed. They were analyzed using polarized light microscopy, and in the case this method had questionable results we used X-ray diffraction. The variables collected were the lithiasis chemical composition (oxalates, phosphates, uric acid, infectious, cystine, mixed, other). Regarding the date of the analysis, the series of cases was grouped into four periods (1980-1989, 1990- 1999, 2000-2009, and 2010-2015), and also by sex and age of the patient. The mean age at diagnosis was 48.32 years (49.37 in men vs 46.53 in women, p=0.005). A male predominance was found (58.7%). Throughout the four decades, the involvement of women has progressively diminished compared to men. Of all the lithiases, the most frequent were those composed of oxalates (43.3%), followed by uric acid (16.9%) and infectious types (10.7%). The uric acid and oxalate lithiases were more common in men than in women (67.4% vs. 32.6% and 59.1% vs. 40.9%, respectively, p<0.001), while the lithiasis of infectious origin was more frequent in women than in men (56.3% vs. 43.7%, p<0.001). Throughout the time period, a trend of increasing oxalic lithiases and decreasing uric acid and phosphate lithiases was observed, as well as an increase of infectious lithiases over the past five years. In our setting, urinary lithiases appear more frequently in males at the end of the fourth decade of life. The most common lithiases are composed of oxalates, and their frequency has increased over time, while uric acid and phosphates lithiases have decreased.


Prefrontal cortex morphometry in abstinent adolescent marijuana users: subtle gender effects.
Adult human studies suggest frontal dysfunction associated with chronic marijuana (MJ) use, but due to continued neuromaturation, adult studies may not generalize to adolescents. This study characterized prefrontal cortex (PFC) morphometry in chronic MJ-using adolescents following 1 month of monitored abstinence. Data were collected from MJ users (n = 16) and controls (n = 16) aged 16-18. Extensive exclusionary criteria included co-morbid psychiatric and neurologic disorders. Substance use and anatomical measures were collected after 28 days of monitored abstinence. PFC volumes were ascertained from manual tracing by reliable raters on high-resolution magnetic resonance images. After controlling for lifetime alcohol use, gender and intracranial volume, MJ users did not differ from controls in PFC volume. However, marginal group-by-gender interactions were observed (P < 0.09): female MJ users demonstrated comparatively larger PFC volumes while male MJ users had smaller volumes compared with same-gender controls. Further, group status and total PFC volume interacted in predicting executive functioning (P < 0.05). Among MJ users, smaller PFC total volume was associated with better executive functioning while the opposite pattern was seen among the controls. These preliminary results indicate that gender may moderate the relationship between MJ use and PFC morphometry. Given the relationship between larger PFC total volumes and poorer executive functioning among MJ users, female MJ users may be at increased risk for neurocognitive consequences. Future research will measure PFC gray and white matter separately and follow boys and girls over adolescence to examine the influence of MJ use on neurodevelopment.


An automated and portable microfluidic chemiluminescence immunoassay for quantitative detection of biomarkers.
Microfluidic platforms capable of automated, rapid, sensitive, and quantitative detection of biomarkers from patient samples could make a major impact on clinical or point-of-care (POC) diagnosis. In this work, we realize an automated diagnostic platform composed of two main components: (1) a disposable, self-contained, and integrated microfluidic chip and (2) a portable instrument that carries out completely automated operations. To demonstrate its potential for real-world application, we use injection molding for mass fabrication of the main components of disposable microfluidic chips. The assembled three-layered chip with on-chip mechanical valves for fluid control consists of (1) a top silicone fluidic layer with embedded zigzag microchannels, reagent reservoirs and a negative pressure port, (2) a middle tinfoil layer with patterned antibody/antigen stripes, and (3) a bottom silicone substrate layer with waste reservoirs. The versatility of the microfluidics-based system is demonstrated by implementation of a chemiluminescence immunoassay for quantitative detection of C-reactive protein (CRP) and testosterone in real clinical samples. This lab-on-a-chip platform with features of quantitation, portability and automation provides a promising strategy for POC diagnosis.


Evaluating tilt angles of membrane-associated helices: comparison of computational and NMR techniques.
A computational method to calculate the orientation of membrane-associated alpha-helices with respect to a lipid bilayer has been developed. It is based on a previously derived implicit membrane representation, which was parameterized using the structures of 46 alpha-helical membrane proteins. The method is validated by comparison with an independent data set of six transmembrane and nine antimicrobial peptides of known structure and orientation. The minimum energy orientations of the transmembrane helices were found to be in good agreement with tilt and rotation angles known from solid-state NMR experiments. Analysis of the free-energy landscape found two types of minima for transmembrane peptides: i), Surface-bound configurations with the helix long axis parallel to the membrane, and ii), inserted configurations with the helix spanning the membrane in a perpendicular orientation. In all cases the inserted configuration also contained the global energy minimum. Repeating the calculations with a set of solution NMR structures showed that the membrane model correctly distinguishes native transmembrane from nonnative conformers. All antimicrobial peptides investigated were found to orient parallel and bind to the membrane surface, in agreement with experimental data. In all cases insertion into the membrane entailed a significant free-energy penalty. An analysis of the contributions of the individual residue types confirmed that hydrophobic residues are the main driving force behind membrane protein insertion, whereas polar, charged, and aromatic residues were found to be important for the correct orientation of the helix inside the membrane.


The mechanism of transport of the multitargeted antifolate (MTA) and its cross-resistance pattern in cells with markedly impaired transport of methotrexate.
MTA (LY231514) is an antifolate that targets multiple folate-dependent enzymes. In this report, MTA transport was characterized in wild-type L1210 cells and variants with impaired membrane transport or polyglutamation. MTA influx via the reduced folate carrier was somewhat faster (approximately 30%) than that for methotrexate (MTX). Unlike MTX, MTA was rapidly polyglutamated in L1210 cells; hence, a folylpoly-gamma-glutamate synthetase-deficient L1210 variant was used to assess net transport and efflux properties. The MTA transmembrane gradient for exchangeable drug was 2.5 times greater than the MTX gradient, attributable primarily to an efflux rate constant 40% that of MTX. No MTA was bound to dihydrofolate reductase. When grown with folic acid, MTX-resistant L1210 variants with mutations in the reduced folate carrier demonstrated cross-resistance to MTA, markedly reduced MTA accumulation, and only a slightly decreased intracellular folate cofactor pool as compared to L1210 cells. However, when 5-formyltetrahydrofolate was the growth substrate, these MTX-resistant cells were less resistant or negligibly resistant to MTA, accumulated more MTA, and had a lower folate pool as compared to L1210 cells. MTA activity and the intracellular folate pool in L1210 cells were inversely related. These data indicate that MTA polyglutamation in L1210 cells is favored by both the generation of high intracellular drug levels and high MTA affinity for FPGS relative to MTX. Cells resistant to MTX because of impaired transport may retain appreciable sensitivity to MTA because of a concurrent reduction in tetrahydrofolate cofactor transport resulting in cellular folate depletion, which diminishes endogenous folate suppression of MTA polyglutamation.


Patient-family agreement on values and preferences for life-sustaining treatment: results of a multicentre observational study.
To quantify agreement between patients and their family members on their own values and preferences for use or non-use of life-sustaining treatments for the patient. Hospitalised patients aged 55 years or older with advanced pulmonary, cardiac, liver disease or metastatic cancer or aged 80 years or older from medical wards at 16 Canadian hospitals and their family members completed a questionnaire including eight items about values related to life-sustaining treatment and a question about preferences for life-sustaining treatments. We recruited a total of 313 patient-family member dyads. Crude agreement between patients and family members about values related to life-sustaining treatment was 42% across all eight items but varied widely: 20% when asking how important it was for the patient to respect the wishes of family members regarding their care; 72% when asking how important it was for the patient to be kept comfortable and suffer as little as possible. Crude agreement on preferences for life-sustaining treatment was 91% (kappa 0.60; 95%CI 0.45 to 0.75) when looking at preferences for cardiopulmonary resuscitation (CPR) versus no CPR but fell to 56% when including all five response options with varying degrees of resuscitative, medical or comfort options (kappa 0.39; 95%CI 0.31 to 0.47). There is appreciable disagreement between seriously ill hospitalised patients and family members in their values and preferences for life-sustaining treatment. Strategies are needed to improve the quality of advance care planning, so that surrogates are better able to honour patient's wishes at the end of life.


Use of restriction fragment polymorphism analysis of rRNA genes to assign species to unknown clinical isolates of oral viridans streptococci.
This study evaluated restriction fragment length polymorphisms of rRNA genes (ribotyping) for genotypic identification of 53 oral isolates classified as "Streptococcus sanguis" by colony morphology. Isolates were from 8-h buccal plaque on lower first permanent molars of 20 subjects. DNA was digested with AatII and hybridized with digoxygenin-labeled cDNA of Escherichia coli 16S and 23S rRNA. Strains were ribotyped again with AlwNI or PvuII on the basis of the presence or absence of a 2,290-bp AatII band. Band patterns were compared with reference ribotypes for Streptococcus gordonii, Streptococcus sanguis, Streptococcus crista, Streptococcus oralis, Streptococcus mitis, and Streptococcus parasanguis strains. Forty-eight isolates could be assigned to a species (22 S. sanguis, 14 S. oralis, 12 S. gordonii). Multiple species were seen in 14 subjects; multiple strains of the same species occurred in 11 subjects. Our findings suggest that ribotyping can be used for genotypic identification of S. sanguis, S. oralis, and S. gordonii isolates.


Coarse-Grained Entanglement and Operator Growth in Anomalous Dynamics.
In two-dimensional Floquet systems, many-body localized dynamics in the bulk may give rise to a chaotic evolution at the one-dimensional edges that is characterized by a nonzero chiral topological index. Such anomalous dynamics is qualitatively different from local-Hamiltonian evolution. Here we show how the presence of a nonzero index affects entanglement generation and the spreading of local operators, focusing on the coarse-grained description of generic systems. We tackle this problem by analyzing exactly solvable models of random quantum cellular automata (QCA) that generalize random circuits. We find that a nonzero index leads to asymmetric butterfly velocities with different diffusive broadening of the light cones and to a modification of the order relations between the butterfly and entanglement velocities. We propose that these results can be understood via a generalization of the recently introduced entanglement membrane theory, by allowing for a spacetime entropy current, which in the case of a generic QCA is fixed by the index. We work out the implications of this current on the entanglement "membrane tension" and show that the results for random QCA are recovered by identifying the topological index with a background velocity for the coarse-grained entanglement dynamics.


Emotional functioning, attachment style, and attributions as predictors of child abuse potential in domestic violence victims.
To explore cognitive and emotional factors that may exacerbate child-abuse potential among domestic violence victims, 80 participants reported on their depression, hopelessness, anxiety, and anger as well as their attachment style and attributional style. Increased emotional difficulties as well as insecure attachment styles were significantly positively correlated with child abuse potential, although depression and anxiety were the strongest predictors. Externalizing blame for the spousal abuse was not associated with abuse risk. Women residing in shelters demonstrated significantly greater abuse risk than those in transitional housing programs, suggesting that greater temporal proximity to the spousal abuse may in part account for the increased abuse potential. Depression and hopelessness, however, appeared particularly relevant to increased abuse risk in domestic violence victims in the transitional housing system. Implications of these findings for working with battered women in terms of their emotional functioning and attachment style are discussed.


Kangaroo care for the preterm infant and family.
Kangaroo care (KC) is the practice of skin-to-skin contact between infant and parent. In developing countries, KC for low-birthweight infants has been shown to reduce mortality, severe illness, infection and length of hospital stay. KC is also beneficial for preterm infants in high-income countries. Cardiorespiratory and temperature stability, sleep organization and duration of quiet sleep, neurodevelopmental outcomes, breastfeeding and modulation of pain responses appear to be improved for preterm infants who have received KC during their hospital stay. No detrimental effects on physiological stability have been demonstrated for infants as young as 26 weeks' gestational age, including those on assisted ventilation. Mothers show enhanced attachment behaviours and describe an increased sense of their role as a mother. The practice of KC should be encouraged in nurseries that care for preterm infants. Information is available to assist in developing guidelines and protocols. Kangaroo care (KC) is the practice of skin-to-skin contact between infant and parent. In developing countries, KC for low-birthweight infants has been shown to reduce mortality, severe illness, infection and length of hospital stay. KC is also beneficial for preterm infants in high-income countries. Cardiorespiratory and temperature stability, sleep organization and duration of quiet sleep, neurodevelopmental outcomes, breastfeeding and modulation of pain responses appear to be improved for preterm infants who have received KC during their hospital stay. No detrimental effects on physiological stability have been demonstrated for infants as young as 26 weeks' gestational age, including those on assisted ventilation. Mothers show enhanced attachment behaviours and describe an increased sense of their role as a mother. The practice of KC should be encouraged in nurseries that care for preterm infants. Information is available to assist in developing guidelines and protocols.


A Descriptive, Cross-sectional Survey of Turkish Nurses' Knowledge of Pressure Ulcer Risk, Prevention, and Staging.
Nurses' knowledge of pressure ulcer (PU) prevention and management is an important first step in the provision of optimal care. To evaluate PU prevention/risk, staging, and wound description knowledge, a descriptive, cross-sectional survey was conducted among nurses working in an acute care Turkish hospital. The survey instrument was a modified and translated version of the Pieper Pressure Ulcer Knowledge Test (PUKT), and its validity and reliability were established. Nurses completed a Personal Characteristics Form, including sociodemographic information and exposure to educational presentations and information about and experience with PUs, followed by the 49-item modified PUKT which includes 33 prevention/risk items, 9 staging items, and 7 wound description items. All items are true/false questions with an I don't know option (scoring: minimum 0, maximum 49). Correct answers received 1 point and incorrect/unknown answers received 0 points. The paper-pencil questionnaires were distributed by 2 researchers to all nurses in the participating hospital and completed by those willing to be included. Responses were analyzed using descriptive statistics. Pearson's correlation test was used to examine the relationship between quantitative variables, and mean scores were compared using the Mann-Whitney U and Kruskal-Wallis tests. Among the 308 participating nurses (mean age 29.5 ± 8.1 [range 19-56] years) most were women (257, 83.4%) with 7.3 ± 7.8 (range 1-36) years of experience. The mean knowledge score for the entire sample was 29.7 ± 6.7 (range 8-42). The overall percentage of correct answers was 60.6% to 61.8% for PU prevention/risk assessment, 60% for wound description, and 56.6% for PU staging. Knowledge scores were significantly (P <.05) higher for participants who attended at least 1 lecture/conference/course on PUs in the last year, read articles/books about PUs, cared for patients with PUs, or believed their patients were at risk for PU development. Most participants (180, 58.4%) scored 60% or more correct; 8 (2.6%) correctly answered 80% or more of the items. The lowest number of correct answers was for the item, "Bunny boots and gel pads relieve pressure on the heels" (22, 7.1%). The results of this study suggest education and experience caring for patients who are at risk for or have a PU affect nurses' knowledge. This study, and additional research examining nurse knowledge, will help the development of much-needed education programs.


MiR-335-5p promotes chondrogenesis in mouse mesenchymal stem cells and is regulated through two positive feedback loops.
Chondrogenic differentiation of mesenchymal stem cells (MSCs) is regulated by many factors and signal pathways, including transcription factors such as Sox9 and microRNAs. MiR-335-5p has been previously reported to regulate osteogenic and adipogenic differentiations of MSCs, but its role in chondrogenic differentiation of MSC remains unknown. In this study, we found that miR-335-5p and its host gene Mest are co-expressed and greatly upregulated during mouse MSCs (mMSCs) chondrogenesis. Overexpression of miR-335-5p in mMSCs increased expression of chondrogenic marker genes. Molecular mechanism explorations revealed that miR-335-5p targets Daam1 and ROCK1, a set of negative regulators of Sox9; Sox9 downregulates the expression of miR-29a and 29b, both negative regulators of Mest expression, thus forming a positive loop from miR-335-5p to Sox9 to Mest/miR-335-5p. In addition, miR-335-5p targets DKK1 during mMSC chondrogenic differentiation to increase β-catenin/TCF activity, which leads to increased level of Mest transcription. These data showed miR-335-5p positively regulates MSC chondrogenesis, and two positive feedback loops are identified for the expression of miR-335-5p and its host gene Mest during the early phase of mMSC chondrogenic differentiation.


Linear relationship between peak and season-long abundances in insects.
An accurate quantitative relationship between key characteristics of an insect population, such as season-long and peak abundances, can be very useful in pest management programs. To the best of our knowledge, no such relationship has yet been established. Here we establish a predictive linear relationship between insect catch Mpw during the week of peak abundance, the length of seasonal flight period, F (number of weeks) and season-long cumulative catch (abundance) A = 0.41MpwF. The derivation of the equation is based on several general assumptions and does not involve fitting to experimental data, which implies generality of the result. A quantitative criterion for the validity of the model is presented. The equation was tested using extensive data collected on captures of male gypsy moths Lymantria dispar (L.) (Lepidoptera: Erebidae) in pheromone-baited traps during 15 years. The model was also tested using trap catch data for two species of mosquitoes, Culex pipiens (L.) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae), in Gravid and BG-sentinel mosquito traps, respectively. The simple, parameter-free equation approximates experimental data points with relative error of 13% and R2 = 0.997, across all of the species tested. For gypsy moth, we also related season-long and weekly trap catches to the daily trap catches during peak flight. We describe several usage scenarios, in which the derived relationships are employed to help link results of small-scale field studies to the operational pest management programs.


Relating magnitudes: the brain's code for proportions.
Whereas much is known about how we categorize and reason based on absolute quantity, data exploring ratios of quantities, as in proportions and fractions, are comparatively sparse. Until recently, it remained elusive whether these two representations of number are connected, how proportions are implemented by neurons and how language shapes this code. New data derived with complementary methods and from different model systems now shed light on the mechanisms of magnitude ratio representations. A coding scheme for proportions has emerged that is remarkably reminiscent of the representation of absolute number. These novel findings suggest a sense for ratios that grants the brain automatic access to proportions independently of language and the format of presentation.


Ox40-Cre-mediated deletion of BRD4 reveals an unexpected phenotype of hair follicle stem cells in alopecia.
BRD4 is a bromodomain extraterminal domain family member and functions primarily as a chromatin reader regulating genes involved in cell-fate decisions. Here, we bred Brd4fl/fl Ox40-Cre mice in which Brd4 was conditionally deleted in OX40-expressing cells to examine the role of BRD4 in regulating immune responses. We found that the Brd4fl/fl Ox40-Cre mice developed profound alopecia and dermatitis, while other organs and tissues were not affected. Surprisingly, lineage-tracing experiments using the Rosa26fl/fl-Yfp mice identified a subset of hair follicle stem cells (HFSCs) that constitutively express OX40, and deletion of Brd4 specifically in such HFSCs resulted in cell death and a complete loss of skin hair growth. We also found that death of HFSCs triggered massive activation of the intradermal γδ T cells, which induced epidermal hyperplasia and dermatitis by producing the inflammatory cytokine IL-17. Interestingly, deletion of Brd4 in Foxp3+ Tregs, which also constitutively express OX40, compromised their suppressive functions, and this, in turn, contributed to the enhanced activation of γδ T cells, as well as the severity of dermatitis and hair follicle destruction. Thus, our data demonstrate an unexpected role of BRD4 in regulating skin follicle stem cells and skin inflammation.


A family of dispersed repeats in Mycobacterium leprae.
The genome of the causative agent of leprosy, Mycobacterium leprae, contains at least 28 copies of a dispersed repetitive sequence, RLEP. From nucleotide sequence analysis it was clear that the RLEP element consists of a 545 bp central domain flanked by a 100 bp left-end and a 44 bp right-end, sometimes associated with a 47 bp extension. The presence of the left and right ends is variable and this allowed three different RLEP configurations to be defined. When the polymerase chain reaction was used to study variation of the central region at least twelve different classes were detected, suggesting that no two RLEP sequences may be identical. Furthermore, they have few features in common with classical bacterial insertion sequences.


Robot assisted laparoscopic partial nephrectomy: a viable and safe option in children.
The safety, benefits and usefulness of laparoscopic partial nephrectomy have been demonstrated in the pediatric population. We describe our technique, and determine the safety and feasibility of robot assisted laparoscopic partial nephrectomy based on our initial experience. We retrospectively reviewed robot assisted laparoscopic partial nephrectomy performed at our institution between 2002 and 2005. The technique was conducted via a transperitoneal approach with the da Vinci Surgical System using standard laparoscopic procedural steps. Clinical indicators of outcomes included estimated blood loss, complications, in hospital narcotic use and length of stay. Robot assisted laparoscopic partial nephrectomy was completed successfully in 9 cases. Mean patient age was 7.2 years and mean followup was 6 months. Mean operative time was 275 minutes and mean estimated blood loss was 49 ml. Operative times improved significantly with experience. Overall patients had a mean hospitalization of 2.9 days and required 1.3 mg morphine per kg. All patients had a normal remaining renal moiety confirmed on Doppler ultrasound. The only complication was an asymptomatic urinoma discovered on ultrasound, which was treated with percutaneous drainage and ultimately resolved. Our initial experience shows the safety and feasibility of robot assisted laparoscopic partial nephrectomy in children. Operative time decreases with experience. The enhanced visualization and dexterity of a robotic system potentially offer improved efficiency and safety over standard laparoscopy. Robot assisted laparoscopy is an option for partial nephrectomy and may become the minimally invasive treatment of choice.


Porosity in posterior composite resins.
The percent pore area and the mean pore size were measured in specimens made of nine brands of composite resin restorative materials. The overall percent porosity ranged from 0.01 to 4.5%. The specimens for the study were prepared in two ways; by condensation or by pressure. Most of the materials, including all those chemically activated, showed more porosity in the condensed than in the pressure specimens. Reversed results emerged for two of the materials, and for another two, the specimens were similar in this respect. The mean pore sizes were smaller in the condensed than in the pressure specimens. Neither mean pore size nor total amount of porosities could be correlated to the curing method of the materials.


Spatial analysis of the distribution of LaCrosse encephalitis in Illinois, using a geographic information system and local and global spatial statistics.
The spatial and temporal distribution of LaCrosse encephalitis cases in Illinois was analyzed using a geographic information system (GIS) and spatial statistics. Case data were obtained from the Illinois Department of Public Health and mapped on the county, town, and address level. Human cases were concentrated in and around the city of Peoria in central Illinois. Local spatial statistics were used to identify hot spots where cases appear to be concentrated in the Peoria region. Several small towns surrounding the city of Peoria appeared as foci where cases were most common. Second-order spatial analysis of the case distribution was conducted on the address level. Cases were clustered within a range of 3.0 km in the city of Peoria. Since most cases appear to be associated with residential (peridomestic) exposure, and since several cases have been reported from neighboring addresses, transmission may be concentrated around specific sites (hardwood ravines, tire piles). The GIS and spatial analysis may be useful in identifying and targeting for intervention potential sites of enzootic transmission.


Methods for the preparation and manufacture of polymeric nanoparticles.
This review summarizes the different methods of preparation of polymer nanoparticles including nanospheres and nanocapsules. The first part summarizes the basic principle of each method of nanoparticle preparation. It presents the most recent innovations and progresses obtained over the last decade and which were not included in previous reviews on the subject. Strategies for the obtaining of nanoparticles with controlled in vivo fate are described in the second part of the review. A paragraph summarizing scaling up of nanoparticle production and presenting corresponding pilot set-up is considered in the third part of the review. Treatments of nanoparticles, applied after the synthesis, are described in the next part including purification, sterilization, lyophilization and concentration. Finally, methods to obtain labelled nanoparticles for in vitro and in vivo investigations are described in the last part of this review.


Tautomerism and electronic spectroscopy of protonated 1- and 2-aminonaphthalene.
Experimental and theoretical investigations of the excited states of protonated 1- and 2-aminonaphthalene are presented. The electronic spectra are obtained by laser induced photofragmentation of the ions captured in a cold ion trap. Using ab initio calculations, the electronic spectra can be assigned to different tautomers which have the proton on the amino group or on the naphthalene moiety. It is shown that the tautomer distribution can be varied by changing the electrospray source conditions, favoring either the most stable form in solution (amino protonation) or that in the gas phase (aromatic ring protonation). Calculations for larger amino-polyaromatics predict that these systems should behave as "proton sponges" i.e. have a proton affinity larger than 11 eV.


Imaging spectrometry for Earth remote sensing.
Imaging spectrometry, a new technique for the remote sensing of the earth, is now technically feasible from aircraft and spacecraft. The initial results show that remote, direct identification of surface materials on a picture-element basis can be accomplished by proper sampling of absorption features in the reflectance spectrum. The airborne and spaceborne sensors are capable of acquiring images simultaneously in 100 to 200 contiguous spectral bands. The ability to acquire laboratory-like spectra remotely is a major advance in remote sensing capability. Concomitant advances in computer technology for the reduction and storage of such potentially massive data sets are at hand, and new analytic techniques are being developed to extract the full information content of the data. The emphasis on the deterministic approach to multispectral data analysis as opposed to the statistical approaches used in the past should stimulate the development of new digital image-processing methodologies.


Utilization of LC-MS/MS analyses to identify site-specific chemical protein adducts in vitro.
Biologically reactive intermediates are formed following metabolism of xenobiotics, and during normal oxidative metabolism. These reactive species are electrophilic in nature and are capable of forming stable adducts with target proteins. These covalent protein modifications can initiate processes that lead to acute tissue injury or chronic disease. Recent advancements in mass spectrometry techniques and data analysis has permitted a more detailed investigation of site-specific protein modifications by reactive electrophiles. Knowledge from such analyses will assist in providing a better understanding of how specific classes of electrophiles produce toxicity and disease progression via site-selective protein-specific covalent modification. Hydroquinone (HQ) is a known environmental toxicant, and its quinone-thioether metabolites, formed via the intermediate generation of 1,4-benzoquinone (1,4-BQ), elicit their toxic response via the covalent modification of target proteins and the generation of reactive oxygen species. We have utilized a model protein, cytochrome c, to guide us in identifying 1,4-BQ- and 1,4-BQ-thioether derived site-specific protein modifications. LC-MS/MS analyses reveals that these modifications occur selectively on lysine and glutamic acid residues of the target protein, and that these modifications occur within identifiable "electrophile binding motifs" within the protein. These motifs are found within lysine-rich regions of the protein and appear to be target sites of 1,4-BQ-thioether adduction. These residues also appear to dictate the nature of post-adduction chemistry and the final structure of the adduct. This model system will provide critical insight for in vivo adduct hunting following exposure to 1,4-BQ-thioethers, but the general approaches can also be extended to the identification of protein adducts derived from other classes of reactive electrophiles.


Genome-wide mapping of RNA Pol-II promoter usage in mouse tissues by ChIP-seq.
Alternative promoters that are differentially used in various cellular contexts and tissue types add to the transcriptional complexity in mammalian genome. Identification of alternative promoters and the annotation of their activity in different tissues is one of the major challenges in understanding the transcriptional regulation of the mammalian genes and their isoforms. To determine the use of alternative promoters in different tissues, we performed ChIP-seq experiments using antibody against RNA Pol-II, in five adult mouse tissues (brain, liver, lung, spleen and kidney). Our analysis identified 38 639 Pol-II promoters, including 12 270 novel promoters, for both protein coding and non-coding mouse genes. Of these, 6384 promoters are tissue specific which are CpG poor and we find that only 34% of the novel promoters are located in CpG-rich regions, suggesting that novel promoters are mostly tissue specific. By identifying the Pol-II bound promoter(s) of each annotated gene in a given tissue, we found that 37% of the protein coding genes use alternative promoters in the five mouse tissues. The promoter annotations and ChIP-seq data presented here will aid ongoing efforts of characterizing gene regulatory regions in mammalian genomes.


Generation and characterization of erythroid cells from human embryonic stem cells and induced pluripotent stem cells: an overview.
Because of the imbalance in the supply and demand of red blood cells (RBCs), especially for alloimmunized patients or patients with rare blood phenotypes, extensive research has been done to generate therapeutic quantities of mature RBCs from hematopoietic stem cells of various sources, such as bone marrow, peripheral blood, and cord blood. Since human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) can be maintained indefinitely in vitro, they represent potentially inexhaustible sources of donor-free RBCs. In contrast to other ex vivo stem-cell-derived cellular therapeutics, tumorigenesis is not a concern, as RBCs can be irradiated without marked adverse effects on in vivo function. Here, we provide a comprehensive review of the recent publications relevant to the generation and characterization of hESC- and iPSC-derived erythroid cells and discuss challenges to be met before the eventual realization of clinical usage of these cells.


Posterior screw fixation in rotationally unstable pelvic ring injuries.
Although the stability of the pelvic ring primarily depends on the integrity of the posterior sacroiliac arch, lateral compression fractures with rotational instability are commonly treated by anterior fixation alone. The objective of the present study was to assess the outcome of patients with these fractures treated by posterior iliosacral screw fixation alone. Patients with rotationally unstable lateral compression fractures of the pelvic ring (Young and Burgess LC I and LC II or AO/Tile B2) treated by percutaneous iliosacral fixation alone were included. Postoperative complications, need for secondary surgery, malunion, secondary fracture displacement and the time to full-weight bearing were documented. Twenty-five patients (13 female, 26 male; age: 56±20 years) were treated by percutaneous screw fixation (14 bilaterally, 11 unilaterally). Mean follow-up was 6±4 months, mean time to full weight bearing 9±3 weeks. Revision surgery was necessary in two patients (8%) due to nerve irritation; an additional anterior stabilisation was needed in two other patients (8%) due to secondary dislocation. Wound infection or motor weakness were not encountered, non-union of the posterior arch did not occur. Non-union of the pubic rami, however, occurred in two patients. The presence of malunion of the pubic rami did not affect the time to full weight bearing. Percutanous iliosacral screw fixation alone is a sufficient technique for the stabilisation of rotationally unstable pelvic fractures with low rates of complications or non-unions. It allows for a minimally invasive treatment thus being a useful option in patients who do not qualify for open anterior fixation.


A Generalized Threat Model for Visual Sensor Networks.
Today, visual sensor networks (VSNs) are pervasively used in smart environments such as intelligent homes, industrial automation or surveillance. A major concern in the use of sensor networks in general is their reliability in the presence of security threats and cyberattacks. Compared to traditional networks, sensor networks typically face numerous additional vulnerabilities due to the dynamic and distributed network topology, the resource constrained nodes, the potentially large network scale and the lack of global network knowledge. These vulnerabilities allow attackers to launch more severe and complicated attacks. Since the state-of-the-art is lacking studies on vulnerabilities in VSNs, a thorough investigation of attacks that can be launched against VSNs is required. This paper presents a general threat model for the attack surfaces of visual sensor network applications and their components. The outlined threats are classified by the STRIDE taxonomy and their weaknesses are classified using CWE, a common taxonomy for security weaknesses.


Real-time estimation of elemental carbon emitted from a diesel engine.
New Mining Safety and Health Administration (MSHA) regulations limit the mass concentration of airborne diesel particulate matter (DPM) or, more specifically, the concentration of elemental carbon (EC), in underground mines. The mine operators are responding by introducing a variety of controls to reduce DPM in the mines, potentially including the evaluation of new maintenance procedures to reduce underground mine vehicle emissions. There is currently a lack of an inexpensive and dependable method to directly measure the DPM concentration emitted from the vehicle tailpipe. To that end, this work demonstrated a simple field portable method for estimating the mass concentration of elemental carbon exiting the tailpipe of a diesel engine using a direct reading photometer. Simultaneous measurements of tailpipe exhaust were made with a Thermo Electron Personal DataRAM 1200 photometer (particulate mass concentration based on light scattering) and by analyzing PM2.5 and PM1.0 samples collected on quartz fiber filters using the National Institute of Occupational Safety and Health (NIOSH) method 5040 (mass concentration of EC via thermal-optical method). Results indicate surprisingly good correlation (R2 = 0.97) of the two methods when the data are adjusted for relative humidity (RH) and corrected using an empirically generated calibration factor. Although preliminary, it may be possible to implement this method in maintenance shops to monitor emission trends and to compare emissions of various vehicles in a fleet. Such data will be useful for fleet planning to meet new air quality standards.


Rapid identification of heterozygous mutations in Drosophila melanogaster using genomic capture sequencing.
One of the key advantages of using Drosophila melanogaster as a genetic model organism is the ability to conduct saturation mutagenesis screens to identify genes and pathways underlying a given phenotype. Despite the large number of genetic tools developed to facilitate downstream cloning of mutations obtained from such screens, the current procedure remains labor intensive, time consuming, and costly. To address this issue, we designed an efficient strategy for rapid identification of heterozygous mutations in the fly genome by combining rough genetic mapping, targeted DNA capture, and second generation sequencing technology. We first tested this method on heterozygous flies carrying either a previously characterized dac(5) or sens(E2) mutation. Targeted amplification of genomic regions near these two loci was used to enrich DNA for sequencing, and both point mutations were successfully identified. When this method was applied to uncharacterized twr mutant flies, the underlying mutation was identified as a single-base mutation in the gene Spase18-21. This targeted-genome-sequencing method reduces time and effort required for mutation cloning by up to 80% compared with the current approach and lowers the cost to <$1000 for each mutant. Introduction of this and other sequencing-based methods for mutation cloning will enable broader usage of forward genetics screens and have significant impacts in the field of model organisms such as Drosophila.


Phase-only modulation with two vertical aligned liquid crystal devices.
Spatial Light Modulators (SLMs) are widely used in several fields of optics such as adaptive optics. SLMs based on Liquid Crystal (LC) devices allow a dynamic and easy representation of two-dimensional phase maps. A drawback of these devices is their elevated cost, preventing a massive use of the technology. We present a more affordable approach based on the serial arrangement of vertical aligned LC devices, with characteristics of phase modulation similar to a widely used parallel aligned LC device. We discuss the peculiarities of the approach, the performance and some potential areas of applications.


Procarbazine--a traditional drug in the treatment of malignant gliomas.
The methylhydrazine derivative Procarbazine (PCZ) as monotherapy or in combination with CCNU and vincristine (PCV) was evaluated in a vast number of clinical trials and is still used in patients with high-grade and low-grade gliomas. The compound is an antineoplastic agent with multiple sites of action. It inhibits incorporation of small DNA precursors, as well as RNA and protein synthesis. PCZ can also directly damage DNA through an alkylation reaction. The drug is not cross-resistant with other mustard-type alkylating agents. As PCZ was in almost all trials used in a combination with CCNU and Vincristin, the efficacy can only be evaluated in the view of the PCV regimen. The published data suggest a role of PCV as a salvage regimen, especially in oligodendroglial tumors; however, well designed studies with high evidence are rare in all entities. This article summarizes the existing data with the goal to define the role of PCZ/PCV in modern neurooncology.


Evaluating adherence to the International Committee of Medical Journal Editors' policy of mandatory, timely clinical trial registration.
To determine whether two specific criteria in Uniform Requirements for Manuscripts (URM) created by the International Committee of Medical Journal Editors (ICMJE)--namely, including the trial ID registration within manuscripts and timely registration of trials, are being followed. Observational study using computerized analysis of publicly available Medline article data and clinical trial registry data. We analyzed a purposive set of five ICMJE founding journals looking at all trial articles published in those journals during 2010-2011, and data from the ClinicalTrials.gov (CTG) trial registry. We measured adherence to trial ID inclusion policy as the percentage of trial journal articles that contained a valid trial ID within the article (journal-based sample). Adherence to timely registration was measured as the percentage of trials that registered the trial before enrolling the first participant within a 60-day grace period. We also examined timely registration rates by year of all phase II and higher interventional trials in CTG (registry-based sample). To determine trial ID inclusion, we analyzed 698 clinical trial articles in five journals. A total of 95.8% (661/690) of trial journal articles included the trial ID. In 88.3% the trial-article link is stored within a structured Medline field. To evaluate timely registration, we analyzed trials referenced by 451 articles from the selected five journals. A total of 60% (272/451) of articles were registered in a timely manner with an improving trend for trials initiated in later years (eg, 89% of trials that began in 2008 were registered in a timely manner). In the registry-based sample, the timely registration rates ranged from 56% for trials registered in 2006 to 72% for trials registered in 2011. Adherence to URM requirements for registration and trial ID inclusion increases the utility of PubMed and links it in an important way to clinical trial repositories. This new integrated knowledge source can facilitate research prioritization, clinical guidelines creation, and precision medicine. The five selected journals adhere well to the policy of mandatory trial registration and also outperform the registry in adherence to timely registration. ICMJE's URM policy represents a unique international mandate that may be providing a powerful incentive for sponsors and investigators to document clinical trials and trial result publications and thus fulfill important obligations to trial participants and society.


Health-Related Quality of Life Outcomes in PARADIGM-HF.
Patients with heart failure and reduced ejection fraction have impaired health-related quality of life (HRQL) with variable responses to therapies that target mortality and heart failure hospitalizations. In PARADIGM-HF trial (Prospective Comparison of ARNI [Angiotensin Receptor-Neprilysin Inhibitor] With ACEI [Angiotensin-Converting-Enzyme Inhibitor] to Determine Impact on Global Mortality and Morbidity in Heart Failure), sacubitril/valsartan reduced morbidity and mortality compared with enalapril. Another major treatment goal is to improve HRQL. Given improvements in mortality with sacubitril/valsartan, this analysis provides comprehensive assessment of impact of therapy on HRQL in survivors only. Patients (after run-in phase) completed disease-specific HRQL using Kansas City Cardiomyopathy Questionnaire (KCCQ) at randomization, 4 month, 8 month, and annual visits. Changes in KCCQ scores were calculated using repeated measures analysis of covariance model that adjusted for treatment and baseline values (principal efficacy prespecified at 8 months). Among the 8399 patients enrolled in PARADIGM-HF, 7623 (91%) completed KCCQ scores at randomization with complete data at 8 months for 6881 patients (90% of baseline). At 8 months, sacubitril/valsartan group noted improvements in both KCCQ clinical summary score (+0.64 versus -0.29; P=0.008) and KCCQ overall summary score (+1.13 versus -0.14; P<0.001) in comparison to enalapril group and significantly less proportion of patients with deterioration (≥5 points decrease) of both KCCQ scores (27% versus 31%; P=0.01). Adjusted change scores demonstrated consistent improvements in sacubitril/valsartan compared with enalapril through 36 months. Change scores in KCCQ clinical summary scores and KCCQ overall summary scores were better in patients treated with sacubitril/valsartan compared with those treated with enalapril, with consistency in most domains, and persist during follow-up beyond 8 months. These findings demonstrate that sacubitril/valsartan leads to better HRQL in surviving patients with heart failure. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01035255.


Somatization: a debilitating syndrome in primary care.
Somatization is a significant problem for clinical medicine. Unlike somatization disorder, which is relatively rare, abridged somatization, a less severe form of somatization, is prevalent in primary care clinics. The authors examined the clinical status and functioning of patients diagnosed with a depression or anxiety disorder comorbid with abridged somatization and compared them with patients diagnosed with a depression or anxiety disorder alone. The authors examined severity of physical functioning and psychopathology in relation to diagnostic status. Patients diagnosed with both abridged somatization and a depression or anxiety disorder were more physically impaired and more anxious than those diagnosed with a depression or anxiety disorder alone. The results suggest that abridged somatization frequently coexists with depression and anxiety and thus complicates the presentation of these disorders.


Mass Spectrometry Imaging with Isomeric Resolution Enabled by Ozone-Induced Dissociation.
Mass spectrometry imaging (MSI) enables the spatial distributions of molecules possessing different mass-to-charge ratios to be mapped within complex environments revealing regional changes at the molecular level. Even at high mass resolving power, however, these images often reflect the summed distribution of multiple isomeric molecules, each potentially possessing a unique distribution coinciding with distinct biological function(s) and metabolic origin. Herein, this chemical ambiguity is addressed through an innovative combination of ozone-induced dissociation reactions with MSI, enabling the differential imaging of isomeric lipid molecules directly from biological tissues. For the first time, we demonstrate both double bond- and sn-positional isomeric lipids exhibit distinct spatial locations within tissue. This MSI approach enables researchers to unravel local lipid molecular complexity based on both exact elemental composition and isomeric structure directly from tissues.


Pilot study of emotional freedom techniques, wholistic hybrid derived from eye movement desensitization and reprocessing and emotional freedom technique, and cognitive behavioral therapy for treatment of test anxiety in university students.
This study explored test anxiety benefits of wholistic hybrid derived from eye movement desensitization and reprocessing and Emotional Freedom Techniques (WHEE), Emotional Freedom Techniques (EFTs), and cognitive behavioral therapy (CBT). Canadian university students with severe or moderate test anxiety participated. A controlled trial of WHEE (n = 5), EFT (n = 5), and CBT (n = 5) was conducted. Standardized anxiety measures included the Test Anxiety Inventory and Hopkins Symptom Checklist-21. Despite small sample size, significant reductions in test anxiety were found for all three treatments. In only two sessions, WHEE and EFT achieved the same benefits as CBT did in five sessions. Participants reported high satisfaction with all treatments. Emotional freedom techniques and WHEE participants successfully transferred their self-treatment skills to other stressful areas of their lives. Both WHEE and EFT show promise as feasible treatments for test anxiety.


Glucose metabolism in patients with schizophrenia treated with olanzapine or quetiapine: a frequently sampled intravenous glucose tolerance test and minimal model analysis.
Clozapine and olanzapine treatment has been associated with insulin resistance in non-obese schizophrenia patients. Much less is known regarding other agents such as quetiapine. The objective of this study was to compare matched olanzapine- and quetiapine-treated schizophrenia patients and normal controls on measures of glucose metabolism. A cross-sectional comparison of quetiapine-treated and olanzapine-treated non-obese (body mass index < 30.0 kg/m2) schizophrenia subjects (DSM-IV) with matched normal controls using a frequently sampled intravenous glucose tolerance test and nutritional assessment was conducted from April 2002 to October 2004. Data from 24 subjects were included in the analysis (7 quetiapine, 8 olanzapine, 9 normal controls). There was a significant difference among groups for fasting baseline plasma glucose concentrations (p = .02), with olanzapine greater than normal controls (p = .01). The insulin sensitivity index (SI) differed significantly among groups (p = .039); olanzapine subjects exhibited significant insulin resistance compared to normal controls (p = .01), but there was no significant difference for quetiapine versus olanzapine (p = .1) or quetiapine versus normal controls (p = .40). SI inversely correlated with quetiapine dose (p = .0001) and waist circumference (p = .03) in quetiapine-treated subjects. Insulin resistance calculated by the homeostasis model assessment of insulin resistance (HOMA-IR) also differed significantly among groups (p = .03). The olanzapine group had a higher HOMA-IR level than normal controls (p = .01). There was a significant difference in glucose effectiveness (SG) among the groups (p = .049). SG was lower in the olanzapine group than in the quetiapine group (p = .03) and in the olanzapine group compared to normal controls (p = .049). Our findings are consistent with our previous report that nonobese olanzapine-treated subjects showed insulin resistance, measured by both HOMA-IR and SI, and reduction in SG. Studies that include larger samples, unmedicated patients, and varying durations of antipsychotic exposure are necessary to confirm these results.


A novel inflammatory role for platelets in sickle cell disease.
The severe pain, ischemia and organ damage that characterizes sickle cell disease (SCD) is caused by vaso-occlusion, which is the blockage of blood vessels by heterotypic aggregates of sickled erythrocytes and other cells. Vaso-occlusion is also a vasculopathy involving endothelial cell dysfunction, leukocyte activation, platelet activation and chronic inflammation resulting in the multiple adhesive interactions between cellular elements. Since platelets mediate inflammation as well as thrombosis via release of pro- and anti-inflammatory molecules, we hypothesized that platelets may play an active inflammatory role in SCD by secreting increased amounts of cytokines. Since platelets have been shown to contain mRNA and actively produce proteins, we also hypothesized that SCD platelets may contain increased cytokine mRNA. In this cross-sectional study, we sought to compare both the quantity of cytokines secreted and the cytokine mRNA content, between SCD and control platelets. We measured the secretion of Th1, Th2, and Th17-related cytokines from platelets in a cohort of SCD patients. We simultaneously measured platelet mRNA levels of those cytokines. Platelets from SCD patients secreted increased quantities of IL-1β, sCD40L, and IL-6 compared to controls. Secretion was increased in patients with alloantibodies. Additionally, mRNA of those cytokines was increased in SCD platelets. Platelets from sickle cell patients secrete increased amounts of inflammatory cytokines, and contain increased cytokine mRNA. These findings suggest a novel immunological role for platelets in SCD vasculopathy, in addition to their thrombotic role, and strengthen the rationale for the use of anti-platelet therapy in SCD.


A generalized method for transfecting root epidermis uncovers endosomal dynamics in Arabidopsis root hairs.
Progress in analysing the cellular functions of many structural proteins has accelerated through the use of confocal microscopy together with transient gene expression. Several methods for transient expression have been developed in the past few years, but their application has seen limited success beyond a few tractable species and tissues. We have developed a simple and efficient method to visualize fluorescent proteins in Arabidopsis root epidermis using co-cultivation of seedlings with Agrobacterium rhizogenes. The method is equally suitable for transient gene expression in other species, including Thellungiella, and can be combined with supporting molecular and biochemical analyses. The method promises significant advantages for study of membrane dynamics, cellular development and polar growth in root hairs without interference in the development of the plant. Since the method targets specifically the root epidermis, it also offers a powerful tool to approach issues of root-rhizosphere interactions, such as ion transport and nutrient acquisition. As a proof of principle, we carried out transfections with fluorescent markers for the plasma membrane (NpPMA2-GFP, Nicotiana plumbaginifolia L. Plasma Membrane H(+)-ATPase 2), the endoplasmic reticulum (YFP-HDEL), and the Golgi apparatus (sialyl transferase-GFP) to trace their distribution in growing Arabidopsis root hairs and epidermis. The results demonstrate that, in Arabidopsis root hairs, movement of the Golgi is faster than previously reported for tobacco leaf epidermal cells, consistent with the high secretory dynamics of the tip growing cell; they show a pattern to the endoplasmic reticulum within the cytoplasm that is more diffuse than found in tobacco leaf epidermis, and they confirm previous findings of a polarized distribution of the endoplasmic reticulum at the tip of growing root hairs.


Adherence of Candida albicans germ tubes to plastic: ultrastructural and molecular studies of fibrillar adhesins.
Germ tubes of Candida albicans produced an additional fibrillar surface layer responsible for enhanced adherence to plastic. The correlation between germination of C. albicans and adherence of germ tubes to a plastic matrix led us to consider the existence of germ tube-specific adhesive components involved in the attachment process. Using concanavalin A-sensitized latex microspheres, we first detected extracellular molecules on the plastic surface after removal of the adherent germ tubes. Electron microscopy confirmed that fibrils of the germ tube involved in cell-substratum interconnections were retained on the plastic surface. Cytochemistry with concanavalin A-gold labeling demonstrated that these fibrillar structures contained mannoproteins. Dithiothreitol and iodoacetamide treatment of washed plastic allowed us to further characterize these fibrillar adhesins. Through analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two components with molecular weights (MWs) of 68,000 and 60,000 were detected on the plastic surface. The 68,000-MW component appeared to be one of the major constituents of the germ tube surface layers. Biosynthetic labeling experiments performed with L-[35S]methionine revealed two additional proteins: a high-MW component (greater than 200,000), and a 200,000-MW component. These four proteins, strongly labeled on the plastic surface and on the germ tube cell wall layers, were in contrast slightly labeled or even nonidentified in the culture supernatant, suggesting their involvement in germ tube adherence.


Managing the epileptic patient.
A review of hospital charts revealed that only 3 (7.5%) of 40 inpatients with a diagnosis of epilepsy had received counseling about the disorder, its treatment, and its social ramifications. Although it is often neglected--as indicated by this small sample--proper and continuing patient education may dispel some of the damaging misconceptions and fears about the disorder and help the epileptic patient lead a full and productive life. Physicians should point out the importance of acceptance of the diagnosis, record keeping by the patient, awareness of precipitating factors, associated socioeconomic problems, and follow-up, stressing the possibility of normal life.


Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) increases human hepatic stellate cell activation.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a halogenated aromatic hydrocarbon that elicits toxicity through the aryl hydrocarbon receptor (AhR). In the liver, gross markers of TCDD toxicity are attributed to AhR activation in parenchymal hepatocytes. However, less is known regarding the consequences of TCDD treatment on non-parenchymal cells in the liver. Hepatic stellate cells (HSCs) are non-parenchymal cells that store vitamin A when quiescent. Upon liver injury, activated HSCs lose this storage ability and instead function in the development and maintenance of inflammation and fibrosis through the production of pro-inflammatory mediators and collagen type I. Reports that TCDD exposure disrupts hepatic retinoid homeostasis and dysregulates extracellular matrix remodeling in the liver led us to speculate that TCDD treatment may disrupt HSC activity. The human HSC line LX-2 was used to test the hypothesis that TCDD treatment directly activates HSCs. Results indicate that exposure to 10nM TCDD almost completely inhibited lipid droplet storage in LX-2 cells cultured with retinol and palmitic acid. TCDD treatment also increased LX-2 cell proliferation, expression of α-smooth muscle actin, and production of monocyte chemoattractant protein-1 (MCP-1), all of which are characteristics of activated HSCs. However, TCDD treatment had no effect on Col1a1 mRNA levels in LX-2 cells stimulated with the potent profibrogenic mediator, transforming growth factor-β. The TCDD-mediated increase in LX-2 cell proliferation, but not MCP-1 production, was abolished when phosphoinositide 3-kinase was inhibited. These results indicate that HSCs are susceptible to direct modulation by TCDD and that TCDD likely increases HSC activation through a multi-faceted mechanism.


Is use of homeopathy associated with poor prescribing in English primary care? A cross-sectional study.
Objectives Prescribing of homeopathy still occurs in a small minority of English general practices. We hypothesised that practices that prescribe any homeopathic preparations might differ in their prescribing of other drugs. Design Cross-sectional analysis. Setting English primary care. Participants English general practices. Main outcome measures We identified practices that made any homeopathy prescriptions over six months of data. We measured associations with four prescribing and two practice quality indicators using multivariable logistic regression. Results Only 8.5% of practices (644) prescribed homeopathy between December 2016 and May 2017. Practices in the worst-scoring quartile for a composite measure of prescribing quality (>51.4 mean percentile) were 2.1 times more likely to prescribe homeopathy than those in the best category (<40.3) (95% confidence interval: 1.6-2.8). Aggregate savings from the subset of these measures where a cost saving could be calculated were also strongly associated (highest vs. lowest quartile multivariable odds ratio: 2.9, confidence interval: 2.1-4.1). Of practices spending the most on medicines identified as 'low value' by NHS England, 12.8% prescribed homeopathy, compared to 3.9% for lowest spenders (multivariable odds ratio: 2.6, confidence interval: 1.9-3.6). Of practices in the worst category for aggregated price-per-unit cost savings, 12.7% prescribed homeopathy, compared to 3.5% in the best category (multivariable odds ratio: 2.7, confidence interval: 1.9-3.9). Practice quality outcomes framework scores and patient recommendation rates were not associated with prescribing homeopathy (odds ratio range: 0.9-1.2). Conclusions Even infrequent homeopathy prescribing is strongly associated with poor performance on a range of prescribing quality measures, but not with overall patient recommendation or quality outcomes framework score. The association is unlikely to be a direct causal relationship, but may reflect underlying practice features, such as the extent of respect for evidence-based practice, or poorer stewardship of the prescribing budget.


Primary Sjogren's syndrome with central nervous system involvement.
To describe the clinical, laboratory, and radiological features of Primary Sjogren's syndrome (PSS) with central nervous system (CNS) involvement. A retrospective case series of 12 female patients with PSS and CNS involvement at King Faisal Specialist Hospital and Research Center, Riyadh, Kingdom of Saudi Arabia from 1991-2009. The diagnosis of PSS is defined by the American-European Diagnostic Criteria. We analyzed the clinical, radiological, and immunological features. The mean age was 40 years (range 16-58 years); all patient were females and presented with active neurological symptoms. The neurological involvement preceded the classic sicca symptoms (33%). Eight patients (66%) presented with myelopathy, 9 patients (75%) had optic neuritis, and the rest had variable neurological signs. Immunological tests (anti-Sjogren's syndrome A and anti-Sjogren's syndrome B) were high in 7 patients (58%). Minor salivary gland biopsy revealed inflammatory cell infiltrate in 11 patients (92%). Brain MRI showed scattered white matter changes in 7 patients (58%). Spine MRI showed multiple foci of hyperintensity in T2-weighted image in 6 patients (50%), and long segment of hyperintensity at the cervical spinal cord in 2 patients (16%). Our findings demonstrate that CNS involvements in PSS have great clinical variability and could precede the classic sicca symptoms by years. Primary Sjogren's syndrome can mimic multiple sclerosis (primary progressive multiple sclerosis or relapsing remitting multiple sclerosis), therefore a screening test for PSS should be considered in suspected cases. A well-defined management protocol awaits studies with larger case numbers.


Randomized phase III trial of paclitaxel/carboplatin with or without PF-3512676 (Toll-like receptor 9 agonist) as first-line treatment for advanced non-small-cell lung cancer.
This phase III study examined efficacy of the synthetic Toll-like receptor 9-activating oligodeoxynucleotide PF-3512676 in combination with standard paclitaxel/carboplatin chemotherapy in patients with advanced non-small-cell lung cancer (NSCLC). Chemotherapy-naive patients with stage IIIB or IV NSCLC were randomly assigned (1:1) to receive up to six courses of paclitaxel/carboplatin (intravenous paclitaxel 200 mg/m(2) and carboplatin at area under the [concentration-time] curve 6 on day 1 of a 3-week cycle) alone (control arm) or in combination with 0.2 mg/kg subcutaneous PF-3512676 on days 8 and 15 (investigational arm). Primary end point was overall survival (OS). Baseline demographics were similar across arms (N = 828). Most patients (88%) had stage IV disease. Median OS and median progression-free survival (PFS) were similar (OS: investigational arm, 10.0 months v control arm, 9.8 months; P = .56; PFS: investigational arm, 4.8 months v control arm, 4.7 months; P = .79). Most commonly reported PF-3512676-related adverse events (AEs) were mild-to-moderate local injection site reactions, pyrexia, and flu-like symptoms. In the investigational arm, grades 3 to 4 AEs, including neutropenia, thrombocytopenia, and anemia, were more frequent, and more patients had one or more sepsis-related AEs versus controls (17 v 3). At first interim analysis, the Data Safety Monitoring Committee recommended study discontinuation because of lack of incremental efficacy and more sepsis-related serious AEs in the PF-3512676 arm. Administration of PF-3512676, but not chemotherapy, was halted. Addition of PF-3512676 to paclitaxel/carboplatin did not improve OS or PFS versus paclitaxel/carboplatin alone for first-line treatment of patients with advanced NSCLC but did increase toxicity. This regimen cannot be recommended for treating patients with advanced NSCLC.


[Clinical features and toxicity of tuberculosis treatment in patients with cancer].
To assess the clinical and epidemiological characteristics of active tuberculosis in patients with malignancy and to assess the influence of TB treatment on cancer management at the National Institute of Neoplastic Diseases from 2008 to 2013. Observational study of TB cases diagnosed by positive sputum microscopy in patients with cancer. Clinical information, evolution, and pathologic information of neoplasia was reviewed. 76 cases of active tuberculosis after being diagnosed with cancer were found. The median age was 51.3 years. Median follow-up was 2.1 years. The most common cancers were acute lymphocytic leukemia (14.5%), for the hematologic cancers; and cancer of the cervix (14.5%), breast (10.5%), and gastric (7.9%) for non-hematological cancers. 27.6% of patients had recurrence of the tumor; TB diagnosis confounded the initial staging by 6.9% and was initially stated as cancer recurrence in 11.1% (breast and colon cancers). The diagnosis of tuberculosis delayed or influenced the dose reduction of the antineoplastic treatment in 11.1% of the cases (acute lymphocytic leukemia and non-Hodgkin lymphoma). 8.3% of patients had toxicity to the TB treatment. Cancer patients may have active tuberculosis infection. The interference effect of diagnosis and treatment of tuberculosis on the assessment of cancer and cancer treatment in our series is minimal.


Child abuse and other traumatic experiences, alcohol use disorders, and health problems in adolescence and young adulthood.
We prospectively examined the health effects of child abuse and other traumatic events, with objective health indicators and consideration of alcohol use disorders (AUD). Adolescents (n = 668) were recruited from clinical and community sources. At baseline, we examined child abuse and other traumas, AUD, health-related symptoms, physical findings, and blood assays. Subjects were assigned to Trauma Classes (TC), including witnessing violence, physical abuse, and sexual abuse. Health outcomes were again determined at 1-year and young adult follow-up. In adolescence, higher TC severity was associated with more health-related symptoms, increased age-adjusted body mass index, and stress-response immune system indices. In adolescence and young adulthood, the relationships between TC and health-related symptoms were mediated by anxiety. AUD was associated with liver injury, and cigarette smoking with heart/lung symptoms. Child abuse predicted persistently elevated health-related symptoms primarily attributable to anxiety, and early signs of liver disease were attributable to AUD.


Abnormal surface distribution of the human asialoglycoprotein receptor in cirrhosis.
Serum concentrations of asialoglycoproteins are increased in cirrhosis. We hypothesized that this increase results from abnormalities in the asialoglycoprotein receptor, which is located on the sinusoidal and lateral membrane of hepatocytes. Therefore we searched for morphological alterations in the distribution of the asialoglycoprotein receptor in human liver, using a light microscopic immunoperoxidase method in autopsy livers. In 24 of 25 (96%) of patients without liver disease, the asialoglycoprotein receptor was located on the sinusoidal and, less prominently, the lateral surface of hepatocytes but not the canalicular surface. In contrast, in 12 of 18 (67%) patients with cirrhosis of various causes, the receptor also was localized strikingly along the canalicular surface, with a corresponding decrease on the sinusoidal and lateral surfaces. We conclude that an abnormal cell-surface distribution of the asialoglycoprotein receptor commonly occurs in cirrhosis. This abnormality might result in impaired clearance of desialylated glycoproteins from plasma.


Effects of impairment in activities of daily living on predicting mortality following hip fracture surgery in studies using administrative healthcare databases.
Impairment in activities of daily living (ADL) is an important predictor of outcomes although many administrative databases lack information on ADL function. We evaluated the impact of ADL function on predicting postoperative mortality among older adults with hip fractures in Ontario, Canada. Sociodemographic and medical correlates of ADL impairment were first identified in a population of older adults with hip fractures who had ADL information available prior to hip fracture. A logistic regression model was developed to predict 360-day postoperative mortality and the predictive ability of this model were compared when ADL impairment was included or omitted from the model. The study sample (N = 1,329) had a mean age of 85.2 years, were 72.8% female and the majority resided in long-term care (78.5%). Overall, 36.4% of individuals died within 360 days of surgery. After controlling for age, sex, medical comorbidity and medical conditions correlated with ADL impairment, addition of ADL measures improved the logistic regression model for predicting 360 day mortality (AIC = 1706.9 vs. 1695.0; c -statistic = 0.65 vs 0.67; difference in - 2 log likelihood ratios: χ(2) = 16.9, p = 0.002). Direct measures of ADL impairment provides additional prognostic information on mortality for older adults with hip fractures even after controlling for medical comorbidity. Observational studies using administrative databases without measures of ADLs may be potentially prone to confounding and bias and case-mix adjustment for hip fracture outcomes should include ADL measures where these are available.


Effect of extracellular Ca++ omission on isolated hepatocytes. I. Induction of oxidative stress and cell injury.
The effect of varying extracellular Ca++ concentration and various antioxidants and metal ion chelators on the viability of isolated rat hepatocytes incubated under 95% O2-5% CO2 was evaluated. Decreasing the extracellular Ca++ concentration from 2 mM to 50 microM or less resulted in a progressive increase in cell injury as determined by lactate dehydrogenase (LDH) leakage. The generation of an oxidative stress, indicated by malondialdehyde formation, occurred before and concomitant with LDH leakage. The antioxidants vitamin E, N,N'-diphenyl-p-phenyl-enediamine, chlorpromazine and promethazine, as well as the iron chelators desferrioxamine and EDTA, all prevented both malondialdehyde production and LDH leakage induced by the absence of extracellular Ca++. A marked loss of cytosolic and mitochondrial glutathione was observed in cells incubated without Ca++, which could be significantly prevented by antioxidants and iron chelators. Similar effects on hepatocyte vitamin E levels also were noted. Addition of the Ca++ chelator ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid to the incubation medium totally prevented LDH leakage and malondialdehyde production and partially ameliorated glutathione and vitamin E loss. Conversely, cell injury resulting from disrupted cellular Ca++ homeostasis after treatment with the Ca++ ionophore A23187 plus 2 mM Ca++ was only slightly inhibited by antioxidants and iron chelators. However, ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid also prevented A23187-induced cell injury. These results indicate that a decrease in extracellular Ca++ generates an oxidative stress in hepatocytes which leads ultimately to severe cell injury. This oxidative stress appears to be a Ca++-dependent phenomenon, precipitating all toxicity by a mechanism distinct from that induced by A23187 treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


Heterogeneous nuclear ribonucleoprotein (hnRNP) K is a component of an intronic splicing enhancer complex that activates the splicing of the alternative exon 6A from chicken beta-tropomyosin pre-mRNA.
Splicing of the chicken beta-tropomyosin exon 6A is stimulated, both in vivo and in vitro, by an intronic pyrimidine-rich element (S4) located 37 nucleotides downstream of exon 6A. Several pyrimidine-rich sequences are able to substitute for the natural S4 enhancer with various stimulatory effects. We show that the different enhancer sequences recruit U1 small nuclear ribonucleoprotein (SnRNP) to the exon 6A 5' splice site, with an efficiency that correlates with the splicing activation. By using RNA affinity and two-dimensional gel electrophoresis, we characterized several proteins that bind to the different enhancer sequences. Heterogeneous nuclear ribonucleoprotein (hnRNP) K and hnRNP I (polypyrimidine track-binding protein, PTB) exhibit a higher level of interaction with the strong enhancer sequences (S4) than with the weakest enhancers. Functional analysis shows that hnRNP K is a component of the enhancer complex that promotes exon 6A splicing through the wild-type S4 sequence. The addition of recombinant hnRNP K to nuclear extracts preincubated with poly(rC) RNA competitor completely restores splicing efficiency to the original level. hnRNP I (PTB) was also found associated with the strong enhancer sequences. Its function in the splicing of exon 6A is discussed.


Assessment of the safety and biodistribution of a regulated AAV2 gene transfer vector after delivery to murine submandibular glands.
Clinical gene transfer holds promise for the treatment of many inherited and acquired disorders. A key consideration for all clinical gene transfer applications is the tight control of transgene expression. We have examined the safety and biodistribution of a serotype 2, recombinant adeno-associated viral (AAV2) vector that encodes a rapamycin-responsive chimeric transcription factor, which regulates the expression of a therapeutic transgene (human erythropoietin [hEpo]). The vector, AAV2-TF2.3w-hEpo (2.5 × 10(7)-2.5 × 10(10) particles), was administered once to a single submandibular gland of male and female mice and mediated hEpo expression in vivo following a rapamycin injection but not in its absence. Control (saline treated) and vector-treated animals maintained their weight, and consumed food and water, similarly. Vector delivery led to no significant toxicological effects as judged by hematology, clinical chemistry, and gross and microscopic pathology evaluations. On day 3 after vector delivery, vector copies were not only abundant in the targeted right submandibular gland but also detected in multiple other tissues. Vector was cleared from the targeted gland much more rapidly in female mice than in male mice. Overall, our results are consistent with the notion that administration of the AAV2-TF2.3w-hEpo vector to salivary glands posed no significant risk in mice.


The roles of housing, financial, and food insecurities in understanding the relationship between childhood neglect and violence in adulthood.
The aim of this study is to determine whether basic housing, financial, and food insecurities in part explain the relationship between childhood neglect and violence as documented in the "cycle of violence" literature. Using a prospective cohort design, neglected children (under the age of 12) with court substantiated histories (1967-1971) in one metropolitan Midwest area and demographically matched non-neglected children were followed into adulthood. Housing, financial, and food insecurities were assessed in 2003-2005 interviews at mean age 41. Official arrest data were used to measure violence ever and from 2003 through 2013. Mediation was tested using probit structural equation modeling. Controlling for age, sex, and race, childhood neglect predicted violent arrests and housing, financial, and food insecurities in middle adulthood. Housing and financial securities predicted violent arrests ever and after 2003, whereas food insecurity only predicted any violent arrest ever. Housing and financial insecurities partially mediated the relationship between childhood neglect and violent criminal behavior. Greater attention and efforts need to focus on providing basic housing, financial, and food support for neglected children to reduce their risk for violent criminal behavior.


Identification of a core bacterial community within the large intestine of the horse.
The horse has a rich and complex microbial community within its gastrointestinal tract that plays a central role in both health and disease. The horse receives much of its dietary energy through microbial hydrolysis and fermentation of fiber predominantly in the large intestine/hindgut. The presence of a possible core bacterial community in the equine large intestine was investigated in this study. Samples were taken from the terminal ileum and 7 regions of the large intestine from ten animals, DNA extracted and the V1-V2 regions of 16SrDNA 454-pyrosequenced. A specific group of OTUs clustered in all ileal samples and a distinct and different signature existed for the proximal regions of the large intestine and the distal regions. A core group of bacterial families were identified in all gut regions with clear differences shown between the ileum and the various large intestine regions. The core in the ileum accounted for 32% of all sequences and comprised of only seven OTUs of varying abundance; the core in the large intestine was much smaller (5-15% of all sequences) with a much larger number of OTUs present but in low abundance. The most abundant member of the core community in the ileum was Lactobacillaceae, in the proximal large intestine the Lachnospiraceae and in the distal large intestine the Prevotellaceae. In conclusion, the presence of a core bacterial community in the large intestine of the horse that is made up of many low abundance OTUs may explain in part the susceptibility of horses to digestive upset.


[Parameters of ovulation in clomiphene citrate treated cycles. The effect of pre-ovulatory administration of oestrogen and human chorionic gonadotropin].
This study was undertaken to evaluate the parameters used for monitoring ovulation in patients on clomiphene citrate treatment for the induction of ovulation. The mean serum oestradiol level for every follicle larger than 17 mm on the day before ovulation was 1 885 pmol/l. This value is significantly higher than the value found in spontaneous cycles. The maximum average pre-ovulatory follicle size was 22 mm, also significantly larger than in spontaneous cycles. The maximum pre-ovulatory cervical mucus score is significantly lower than that found in spontaneous cycles. The average midluteal serum progesterone and oestradiol values were significantly higher than in spontaneous cycles. These differences should be taken into consideration in interpreting the results of patients treated with clomiphene citrate for ovulation abnormalities and before in vitro fertilization. Seven patients with a healthy cervix had poor cervical mucus scores in spite of high pre-ovulatory serum oestradiol levels; scores did not improve with administration of exogenous oestrogen, but became optimal with subsequent human menopausal gonadotrophin treatment. Poor cervical mucus despite ovulation and adequate pre-ovulatory serum oestradiol levels in patients on clomiphene treatment should therefore be considered as an indication for gonadotrophin treatment. Pre-ovulatory administration of human chorionic gonadotrophin had no effect in increasing midluteal serum progesterone and oestradiol levels.


Inhalation of hydrogen gas attenuates airway inflammation and oxidative stress in allergic asthmatic mice.
Asthma is a worldwide common chronic airway disease that cannot be cured and results in the huge burden in public health. Oxidative stress was considered an important mechanism in the pathogenesis of asthma. Hydrogen gas been demonstrated to function as a novel antioxidant and exert therapeutic antioxidant activity in a number of diseases and the function of this nontoxic gas in asthma was unclear. The purpose of the study aims to examine the effect of inhalation hydrogen gas on the pathophysiology of a mouse model of asthma. A murine model of ovalbumin (OVA)-induced allergic airway inflammation was used in this study. Briefly, Mice were sensitized to ovalbumin and received inhalation of 67% high concentration of hydrogen gas for 60 min once a day for 7 consecutive days after OVA or PBS challenge respectively. Lung function was assessed in the apparatus with 4 channels of biological signal system. Morphology and goblet cell hyperplasia were stained by H/E and Periodic acid-Schiff staining. Cytologic classification in the bronchial alveolar lavage fluid (BALF) was analyzed by Wright Giemsa staining. Serum, BALF and lung tissue were collected for biochemical assay. One-way analysis of variance (ANOVA) was used to determine statistical significance between groups. Multiple comparisons were made by Bonferroni's Multiple Comparison Test by using GraphPad Prism 5 software. Inhalation of hydrogen gas abrogated ovalbumin-induced the increase in lung resistance. Concomitantly, the asthmatic mice showed severe inflammatory infiltration and goblet cell hyperplasia which were reversed by hydrogen gas inhalation. Hydrogen gas inhalation reduced significantly the number of total cells, eosinophils and lymphocytes in BALF. Increased level of IL-4, IL-13, TNF-α and CXCL15 in the BALF and IL-4 in the serum were decreased significantly after inhalation. Hydrogen gas inhalation markedly upregulated the activity of decreased superoxide dismutase and significantly attenuated the increased level of malondialdehyde and myeloperoxidase. Hydrogen gas inhalation improves lung function and protects established airway inflammation in the allergic asthmatic mice model which may be associated with the inhibition of oxidative stress process. This study provides a potential alternative therapeutic opportunity for the clinical management of asthma.


Supervised classification enables rapid annotation of cell atlases.
Single-cell molecular profiling technologies are gaining rapid traction, but the manual process by which resulting cell types are typically annotated is labor intensive and rate-limiting. We describe Garnett, a tool for rapidly annotating cell types in single-cell transcriptional profiling and single-cell chromatin accessibility datasets, based on an interpretable, hierarchical markup language of cell type-specific genes. Garnett successfully classifies cell types in tissue and whole organism datasets, as well as across species.