Influence of rumen protein degradability and supplementation frequency on performance and nitrogen use in ruminants consuming low-quality forage: cow performance and efficiency of nitrogen use in wethers.
Two studies were conducted to determine the influence of CP degradability and supplementation frequency (SF) on ruminant performance and N efficiency. Treatments included an unsupplemented control (CON) and degradable intake protein (DIP; 82% of CP) or undegradable intake protein (UIP; 60% of CP) provided daily, every 3 d, or every 6 d. Seven wethers (36+/-1 kg BW) were used in the digestibility study with DIP and UIP treatments formulated to meet CP requirements. Eighty-four Angus x Hereford cows (512+/-42 kg BW) in the last third of gestation were used for the performance study. The DIP treatments were calculated to provide 100% of the DIP requirement and UIP treatments were provided on an isonitrogenous basis compared with DIP. Basal diets consisted of low-quality (5% CP) meadow hay. Forage DMI and N intake by lambs decreased (P < 0.05) linearly as SF decreased. Additionally, DMI, OM intake, N retention, N digestibility, and digested N retained were greater (P < 0.01) for supplemented wethers than for controls with no difference due to crude protein degradability. Nitrogen balance, DMI, and OM intake decreased linearly (P < 0.05) as SF decreased. Plasma urea (PU; mM) was measured over a 6-d period and supplemented lambs had increased (P < 0.01) PU compared with CON. Plasma urea linearly decreased (P < 0.01) as SF decreased. Pre- and postcalving (within 14 d and 24 h of calving, respectively) cow weight and body condition score change were more positive (P < 0.05) for supplemented groups than for controls. Results suggest CP supplements consisting of 20 to 60% UIP can be effectively used by ruminants consuming low-quality forage without adversely affecting N efficiency and animal performance, even when provided as infrequently as once every 6 d.


Hybridization and emergence of virulence in opportunistic human yeast pathogens.
Hybridization between different species can result in the emergence of new lineages and adaptive phenotypes. Occasionally, hybridization in fungal organisms can drive the appearance of opportunistic lifestyles or shifts to new hosts, resulting in the emergence of novel pathogens. In recent years, an increasing number of studies have documented the existence of hybrids in diverse yeast clades, including some comprising human pathogens. Comparative and population genomics studies performed on these clades are enabling us to understand what roles hybridization may play in the evolution and emergence of a virulence potential towards humans. Here we survey recent genomic studies on several yeast pathogenic clades where hybrids have been identified, and discuss the broader implications of hybridization in the evolution and emergence of pathogenic lineages. © 2017 The Authors. Yeast published by John Wiley & Sons, Ltd.


Detection of Chlamydia trachomatis infections by polymerase chain reaction in asymptomatic pregnant women with special reference to the utility of the pooling of urine specimens.
Genital Chlamydia trachomatis (CT) infections are one of the most prevalent sexually transmitted infections across the world. In pregnant women, if not detected and treated early, these may result in poor pregnancy outcomes and complications. The present study was aimed to screen CT infections from first void urine (FVU) samples of asymptomatic pregnant women using molecular methods. The secondary objective was to evaluate cost-effectiveness in pooling FVU samples for their diagnostic application. FVU samples were collected from 1000 asymptomatic pregnant women over a period of three years. Pooling was done by including five specimens in one pool in the amount of 10 μl and subjected to polymerase chain reaction (PCR) and further confirmed by direct fluorescent antibody assay (DFA). The age of study participants ranged from 18 to 43 yr with the median±standard deviation of 26±3.84 yr. Majority of positive participants were younger than 25 years. A total of 200 pools were prepared and 20 of these were PCR positive. When individual specimen in 20 positive pools was tested, 20 PCR-positive specimens were identified from 19 pools, of which 16 were positive by DFA. Thus, CT was detected in 1.6 per cent asymptomatic pregnant women in India and pooling strategy resulted in 70 per cent reduction in a number of tests performed. Our study detected C. trachomatis infection in 1.6 per cent asymptomatic pregnant women, and pooling of FVU specimens for PCR testing was found to be a cost-saving strategy in comparison to testing individual samples. Further evaluation and studies on the bigger sample size are warranted to validate these results.


Robotic systems in spine surgery.
Surgical robotic systems have been available for almost twenty years. The first surgical robotic systems were designed as supportive systems for laparoscopic approaches in general surgery (the first procedure was a cholecystectomy in 1987). The da Vinci Robotic System is the most common system used for robotic surgery today. This system is widely used in urology, gynecology and other surgical disciplines, and recently there have been initial reports of its use in spine surgery, for transoral access and anterior approaches for lumbar inter-body fusion interventions. SpineAssist, which is widely used in spine surgery, and Renaissance Robotic Systems, which are considered the next generation of robotic systems, are now FDA approved. These robotic systems are designed for use as guidance systems in spine instrumentation, cement augmentations and biopsies. The aim is to increase surgical accuracy while reducing the intra-operative exposure to harmful radiation to the patient and operating team personnel during the intervention. We offer a review of the published literature related to the use of robotic systems in spine surgery and provide information on using robotic systems.


Bactericidal activities of peptide antibiotics against multidrug-resistant Enterococcus faecium.
Multidrug-resistant Enterococcus faecium has emerged as a serious pathogen for which no effective therapy has been established. In this report, we describe the activities of two peptide antibiotics, ramoplanin and daptomycin, against 15 isolates of E. faecium resistant to vancomycin, ampicillin, and aminoglycosides using time-kill experiments. Both antibiotics were rapidly bactericidal when tested in broth; however, the addition of 50% serum resulted in significant regrowth. The combination of ampicillin with either ramoplanin or daptomycin largely prevented this regrowth. These peptide antibiotics showed good activity against these pathogens. While the development of daptomycin has been halted, ramoplanin may hold promise for the therapy of multidrug-resistant E. faecium, especially when combined with ampicillin.


Impact Evaluation of Training Natural Leaders during a Community-Led Total Sanitation Intervention: A Cluster-Randomized Field Trial in Ghana.
We used a cluster-randomized field trial to evaluate training natural leaders (NLs) as an addition to a community-led total sanitation (CLTS) intervention in Ghana. NLs are motivated community members who influence their peers' behaviors during CLTS. The outcomes were latrine use and quality, which were assessed from surveys and direct observation. From October 2012, Plan International Ghana (Plan) implemented CLTS in 60 villages in three regions in Ghana. After 5 months, Plan trained eight NLs from a randomly selected half of the villages, then continued implementing CLTS in all villages for 12 more months. The NL training led to increased time spent on CLTS by community members, increased latrine construction, and a 19.9 percentage point reduction in open defecation (p < 0.001). The training had the largest impact in small, remote villages with low exposure to prior water and sanitation projects, and may be most effective in socially cohesive villages. For both interventions, latrines built during CLTS were less likely to be constructed of durable materials than pre-existing latrines, but were equally clean, and more often had handwashing materials. CLTS with NL training contributes to three parts of Goal 6 of the Sustainable Development Goals: eliminating open defecation, expanding capacity-building, and strengthening community participation.


Dual Versus Mono Antiplatelet Therapy in Large Atherosclerotic Stroke.
Background and Purpose- Two large-scale randomized controlled trials of recurrent stroke prevention suggest that dual antiplatelet therapy with clopidogrel plus aspirin is beneficial for prevention of subsequent ischemic events. There is a paucity of data, however, on the efficacy or effectiveness of such an approach in the treatment of stroke patients with symptomatic large artery atherosclerotic occlusive disease. Methods- We used a multicenter stroke registry database (Clinical Research Collaboration for Stroke in Korea) to analyze acute ischemic stroke patients due to large artery atherosclerotic occlusive disease who were treated with aspirin alone or combination of clopidogrel and aspirin from May 2008 to May 2015. The results were analyzed by intention-to-treat, per-protocol, and as-treated methodologies. The primary end point was the 1-year composite outcome of stroke recurrence, myocardial infarction, and all-cause death. To balance the differences between groups, a frailty model using propensity scores and inverse probability of treatment weighting was used. Results- A total of 5934 patients with symptomatic large artery atherosclerotic occlusive disease were treated either with clopidogrel plus aspirin (n=2903, 49%) or aspirin (n=3031, 51%). The frequency of the primary outcome was 12% (n=353) in the clopidogrel-aspirin group and 14% (n=410) in the aspirin group. The hazards of the primary outcome with combination over aspirin only were significantly reduced in the per-protocol and as-treated analyses (hazard ratio, 0.71; 95% CI, 0.57-0.88; P=0.002 and hazard ratio, 0.81; 95% CI, 0.69-0.96; P=0.02, respectively), but there was borderline significance in the intention-to-treat analysis (hazard ratio, 0.86; 95% CI, 0.74-1.01; P=0.06). Combination therapy was beneficial for all-cause death in all analyses but did not reduce recurrent stroke. Conclusions- Compared with patients receiving aspirin monotherapy, the primary outcome seemed to occur less frequently in patients receiving dual antiplatelet therapy, which is explained mainly by the decrease of all-cause death. Since this is a nonrandomized, retrospective, observational study, our study should be cautiously interpreted.


Symmetrical serotonin release during asymmetrical slow-wave sleep: implications for the neurochemistry of sleep-waking states.
On land, fur seals predominately display bilaterally synchronized electroencephalogram (EEG) activity during slow-wave sleep (SWS), similar to that observed in all terrestrial mammals. In water, however, fur seals exhibit asymmetric slow-wave sleep (ASWS), resembling the unihemispheric slow-wave sleep of odontocetes (toothed whales). The unique sleeping pattern of fur seals allows us to distinguish neuronal mechanisms mediating EEG changes from those mediating behavioral quiescence. In a prior study we found that cortical acetylcholine release is lateralized during ASWS in the northern fur seal, with greater release in the hemisphere displaying low-voltage (waking) EEG activity, linking acetylcholine release to hemispheric EEG activation (Lapierre et al. 2007). In contrast to acetylcholine, we now report that cortical serotonin release is not lateralized during ASWS. Our data demonstrate that bilaterally symmetric levels of serotonin are compatible with interhemispheric EEG asymmetry in the fur seal. We also find greatly elevated levels during eating and hosing the animals with water, suggesting that serotonin is more closely linked to bilateral variables, such as axial motor and autonomic control, than to the lateralized cortical activation manifested in asymmetrical sleep.


Building a comprehensive genomic program for hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is the most common primary liver cancer, causing approximately 660,000 deaths worldwide annually. The preferred treatment of HCC is surgical resection or orthotopic liver transplantation (OLT) for patients meeting specific criteria. For patients outside these criteria, options are limited and include medical therapy, radiofrequency ablation, chemoembolization, or palliative measures, and these result in poor outcomes. Various centers at Baylor are elucidating the genomics of HCC to improve treatment options, with a focus on three etiologies: hepatitis C virus, hepatitis B virus, and non-viral. Through collaborative efforts, we have established an effective specimen biobanking protocol, and we are using several techniques to analyze HCC, including whole genome sequencing, whole exome sequencing, gene-specific analysis, gene expression, and epigenetic analysis. We have completed whole genome sequencing on two patient samples, whole exome sequencing on 47 patient samples, gene-specific analysis on 94 patient samples, gene expression on 4 patient samples, and epigenetic analysis on 1 patient sample. We hope to use these results to define novel genetic therapeutic strategies that may work in conjunction with surgical approaches to improve long-term patient and graft survival rates in patients with HCC. We also aim to provide a functional framework of a comprehensive program for genomic analysis that may be imitated by other institutions and for other tumors in the global quest toward personalized genomic medicine.


Confusing action and imagination: action source monitoring in individuals with schizotypal traits.
To explore whether schizotypal traits may undermine source monitoring for actions, 67 undergraduate participants (21 men) completed the Schizotypal Personality Scale and were then given a source-monitoring task in which some specific acts had to be performed, whereas others only had to be imagined. Next, participants had to complete an old-new recognition task and ascribe the source, i.e., whether they had performed or only imagined the items (i.e., source monitoring). Participants also completed a working memory capacity task (i.e., Operation Span task). We found that the higher the Schizotypal Personality Scale scores, the poorer recognition and source attribution scores. Relative to participants with low levels of schizotypal traits (i.e., controls), those with higher levels of schizotypal traits more often falsely claimed to have performed actions when in fact they had only imagined them. Although participants high and low in schizotypical traits did not differ in their working memory capacity, poor working memory capacity was related to source misattribution (i.e., increase false alarms). The present findings indicate that schizotypal traits undermine source monitoring for action in a healthy population.


Dipeptidyl peptidase IV (DPP-IV) inhibitory properties of a camel whey protein enriched hydrolysate preparation.
Camel milk proteins contain dipeptidyl peptidase IV (DPP-IV) inhibitory peptides. A camel whey protein concentrate (WPC, 44.7 ± 3.4% (w/w) protein) was prepared and subsequently hydrolysed with trypsin at different temperatures, enzyme to substrate (E:S) ratios and hydrolysis times yielding fifteen hydrolysates (H1-H15). Their DPP-IV half maximal inhibitory concentrations (IC50) ranged from 0.55 ± 0.05 to 1.52 ± 0.16 mg L-1 for H8 and H6, respectively. E:S was the only factor having a significant effect on the DPP-IV IC50 value (p < 0.05). Relatively potent α-lactalbumin-derived DPP-IV inhibitory peptides (LAHKPL and ILDKEGIDY) were detected in selected hydrolysates. Additionally, three potent β-CN-derived peptides, VPV, YPI and VPF having DPP-IV IC50 values of 6.6 ± 0.5, 35.0 ± 2.0 and 55.1 ± 5.8 µM, respectively, were identified. After IPI, VPV is the second most potent DPP-IV inhibitory peptide identified to date, which supports the role of camel milk as an antidiabetic agent.


Analysis, advice, and congressional leadership: the Physician Payment Review Commission and the politics of Medicare.
In 1985, Congress established the Physician Payment Review Commission (PPRC) to help formulate changes in the system used by Medicare to pay for physician services. The recommendations of the PPRC and subsequent legislative action led to fundamental reform. As a new type of advisory body, the PPRC enabled Congress to establish an agenda for physician payment reform and set it into law despite initial resistance from the executive branch. Four key factors contributed to the influence of the commission: (1) an institutional design that enhanced and integrated congressional policy formulation; (2) the quality of the information generated for legislative deliberation; (3) the open, consensual process the commission used to translate that information into policy recommendations; and (4) the strategic packaging of the proposals for reform. In the process leading to enactment of the new payment system, the commission skillfully bridged the traditionally segmented roles of neutral analyst and political advisor for legislators pursuing Medicare reform. Implementation of physician payment reform has been largely an administrative responsibility, in which the PPRC has played a minimal role. The complexity and ambiguity of some of the legislative provisions have left room for administrative officials and interest groups to maneuver according to their priorities. Thus, despite congressional efforts to design a tightly controlled system, a considerable amount of work remains to assure its technical and political success.


Innervation of footpads of normal and mutant mice lacking sweat glands.
Footpads of normal adult mice are innervated by sympathetic and sensory fibers. The sympathetic fibers associated with sweat glands contain acetylcholinesterase and immunoreactivity for vasoactive intestinal peptide. Although catecholamine histofluorescence is absent, the gland innervation exhibits immunoreactivity for tyrosine hydroxylase. A distinct population of sympathetic fibers, which possess catecholamines and neuropeptide Y as well as tyrosinehydroxylase immunoreactivity, innervates blood vessels. Sensory fibers containing immunoreactivity for substance P and calcitonin gene-related peptide course beneath the epidermis and some form endings in it. Treatment of neonatal mice with the adrenergic neurotoxin, 6-hydroxydopamine, results in loss of sympathetic innervation of sweat glands and blood vessels, permits growth of sensory axons into sweat glands, but does not alter the peptidergic sensory innervation of the dermis and epidermis. Three mouse mutations, Tabby (Ta), crinkled (cr), and downless (dl), disrupt the interactions between the mesenchyme and epidermis that are required for normal development of specific epidermal derivatives, including sweat glands. The sympathetic innervation of blood vessels and sensory innervation of footpad skin of the three mutant mice that lack sweat glands is indistinguishable from normal. The sympathetic fibers that normally innervate sweat glands, however, are not present. These results indicate that in the absence of their normal target, the sympathetic fibers that innervate sweat glands are lacking. Furthermore, they suggest that, although sensory fibers may sprout into sympathetic targets in the footpad, the domains occupied by sensory fibers are not normally accessible to sympathetic axons.


Effective dipolar polarizability of amorphous arrays of size-dispersed nanoparticles.
Inhomogeneity of nanoparticle size, shape, and distribution is ubiquitous and inherent in fabricated arrays or may be a deliberate attempt to engineer the optical response. It leads to a spread of polarizabilities of interacting elements and phases of scattered light, and quantitative understanding of these effects is important. Focusing on random/amorphous arrays of optical antennas, we combine T-matrix calculations and an analytical approach based on an effective dipolar polarizability within a film of dipoles framework to quantify the spectral response as a function of the particle inhomogeneity and stochastic clustering. The interplay of position-dependent stochastic coupling and size distribution of antennas determines the optical properties of such arrays as a function of mean/standard deviation of diameter and minimum separation. The resonance wavelength, amplitude, and scattering-to-absorption ratio exhibit oscillations around their size-averaged values with periods and amplitudes given by average structural factors.


Mapping the structure of folding cores in TIM barrel proteins by hydrogen exchange mass spectrometry: the roles of motif and sequence for the indole-3-glycerol phosphate synthase from Sulfolobus solfataricus.
To test the roles of motif and amino acid sequence in the folding mechanisms of TIM barrel proteins, hydrogen-deuterium exchange was used to explore the structure of the stable folding intermediates for the of indole-3-glycerol phosphate synthase from Sulfolobus solfataricus (sIGPS). Previous studies of the urea denaturation of sIGPS revealed the presence of an intermediate that is highly populated at approximately 4.5 M urea and contains approximately 50% of the secondary structure of the native (N) state. Kinetic studies showed that this apparent equilibrium intermediate is actually comprised of two thermodynamically distinct species, I(a) and I(b). To probe the location of the secondary structure in this pair of stable on-pathway intermediates, the equilibrium unfolding process of sIGPS was monitored by hydrogen-deuterium exchange mass spectrometry. The intact protein and pepsin-digested fragments were studied at various concentrations of urea by electrospray and matrix-assisted laser desorption ionization time-of-flight mass spectrometry, respectively. Intact sIGPS strongly protects at least 54 amide protons from hydrogen-deuterium exchange in the intermediate states, demonstrating the presence of stable folded cores. When the protection patterns and the exchange mechanisms for the peptides are considered with the proposed folding mechanism, the results can be interpreted to define the structural boundaries of I(a) and I(b). Comparison of these results with previous hydrogen-deuterium exchange studies on another TIM barrel protein of low sequence identify, alpha-tryptophan synthase (alphaTS), indicates that the thermodynamic states corresponding to the folding intermediates are better conserved than their structures. Although the TIM barrel motif appears to define the basic features of the folding free energy surface, the structures of the partially folded states that appear during the folding reaction depend on the amino acid sequence. Markedly, the good correlation between the hydrogen-deuterium exchange patterns of sIGPS and alphaTS with the locations of hydrophobic clusters defined by isoleucine, leucine, and valine residues suggests that branch aliphatic side-chains play a critical role in defining the structures of the equilibrium intermediates.


Decision-making process for choosing an elective cesarean delivery among primiparas in Taiwan.
A significant proportion of cesarean deliveries in Taiwan were without medical indications and/or on maternal request. The purpose of this study was to understand the decision-making process of choosing an elective cesarean delivery (ELCD) among primiparas in Taiwan. This qualitative exploratory study was guided by grounded theory. Data were collected through in-depth interviews with 20 primiparous women, 15 of whom chose ELCD. Verbatim transcriptions were analyzed using constant comparative analysis and methods of open, axial, and selective coding. The core category that describes the process of ELCD decision making among primiparas is "controlling risks of childbirth and ensuring well-being." The decision process can be divided into three phases: (1) Pre-decision: risk perception, including two subcategories, negative pre-existing ideas about normal spontaneous delivery (NSD) caused worry, and desire for ELCD as the solution; (2) In-decision: risk assessment. Women proactively collected information about NSD and cesarean delivery, and then weighed the personal risk between NSD and ELCD. Risk assessment focused on seven dimensions including safety, health, comfort, efficiency, feminine charms, time and economy; (3) Post-decision: marching onward fearlessly, including two subcategories, belief in ELCD and stress relief, and persuading stakeholders to agree and gaining the required resources for ELCD. At different phases of decision-making, women revealed different concerns, though the sequence of phases was not necessarily unidirectional. Health care providers should be aware of the decision phase and intervene at appropriate times. It is difficult to change a women's decision to have ELCD in the post-decision phase.


Evaluation of the Nonpathogenic Agrobacterium vitis Strain ARK-1 for Crown Gall Control in Diverse Plant Species.
The nonpathogenic strain of Agrobacterium (=Rhizobium) vitis ARK-1 is a biological agent able to effectively control grapevine crown gall. In this study, treating apple, Japanese pear, peach, rose, and tomato by soaking the roots in a cell suspension of ARK-1 before planting into soil infected with tumorigenic Agrobacterium spp. reduced the number of plants developing crown gall tumors. Meta-analyses of the results from six field trials of apple, four field trials of Japanese pear, and four field trials of peach, from 2010 to 2013, showed integrated risk ratio (IRR) after treatment with ARK-1 to be 0.38 for apple crown gall, 0.16 for Japanese pear crown gall, and 0.20 for peach crown gall, indicating that the disease incidence was significantly reduced by ARK-1 treatment. Meta-analyses of the results from three greenhouse trials of rose and three greenhouse trials of tomato showed IRR after treatment with ARK-1 to be 0.29 for rose crown gall and 0.16 for tomato crown gall, indicating that the disease incidence was significantly reduced by ARK-1 treatment. These results indicated that control by ARK-1 covers five different species of host plants and tumorigenic (Ti) strains of Agrobacterium species.


Intracellular targets in heme protein-induced renal injury.
We examined two potential intracellular targets in the glycerol model of acute renal failure, namely, the mitochondrion and the nucleus. Within three hours, alterations in mitochondrial function are already apparent. With either glutamate/malate or succinate/rotenone, state 3 and uncoupled respirations were decreased at three hours, and at 24 hours, such decrements were quite pronounced; in the presence of glutamate/malate, state 2 respiration was also depressed at 24 hours, while with succinate/rotenone state 2 was increased. Marked ultrastructural changes were observed in mitochondria studied at three hours, including the novel finding of degenerate mitochondria in autophagic vacuoles. Since the heme content in mitochondria was increased some tenfold within three hours, mitochondrial function was studied after exposure to concentrations of heme that reproduced such contents of heme: mitochondria initially displayed increased respiration, and subsequently, a persistent decline in oxygen consumption until oxygen consumption was virtually undetectable. With higher concentrations of heme, the early increase in oxygen consumption was blunted and the progressive decline in oxygen consumption was hastened. The antioxidant iron chelator, deferoxamine, prevented the early rise in oxygen consumption but did not prevent or delay the subsequent decline. We also assessed nuclear damage as a potential lesion in the glycerol model. DNA laddering was not observed at any time point. At 3 and 24 hours there was DNA injury by the TUNEL technique in the distal nephron but not in the proximal nephron. The 8-hydroxydeoxyguanosine/deoxyguanosine content was increased in the glycerol kidneys at 24 hours but not at three hours. At neither time point was evidence of apoptosis observed by light or electron microscopy. In studies undertaken in cell culture models, heme, at concentrations of 10 microM, failed to evince any such changes in LLC-PK1 cells, a cell line from the proximal tubule, or in MDCK cells, a cell line derived from the distal tubule. At concentrations of 50 microM, heme induced approximately 20% positivity in MDCK cells but none in LLC-PK1 cells by the TUNEL technique. We conclude that mitochondria and nuclei are prominent targets for injury in the glycerol model of acute renal failure. The presence of TUNEL-positive cells in the distal nephron but not at proximal sites in vivo underscores the increasing appreciation of the distinct responses of these nephron sites to nephrotoxic insults.


Direct estimation of differential networks.
It is often of interest to understand how the structure of a genetic network differs between two conditions. In this paper, each condition-specific network is modeled using the precision matrix of a multivariate normal random vector, and a method is proposed to directly estimate the difference of the precision matrices. In contrast to other approaches, such as separate or joint estimation of the individual matrices, direct estimation does not require those matrices to be sparse, and thus can allow the individual networks to contain hub nodes. Under the assumption that the true differential network is sparse, the direct estimator is shown to be consistent in support recovery and estimation. It is also shown to outperform existing methods in simulations, and its properties are illustrated on gene expression data from late-stage ovarian cancer patients.


Complex N-glycans in Mgat1 null preimplantation embryos arise from maternal Mgat1 RNA.
Mice with a null mutation in the Mgat1 gene lack N-acetyl-glucosaminyltransferase I (GlcNAc-TI; EC 188.8.131.52), and die at mid-gestation. This result suggested that development of Mgat1-/- blastocysts and their implantation could occur in the absence of complex and hybrid N-glycans. However, inner cell mass of all blastocysts from several Mgat1 +/- heterozygous crosses bind the lectin E-PHA, indicating that Mgat1 null mutant blastocysts are able to synthesize complex N-glycans (Campbell et al., (1995) Glycobiology, 5, 535-543). In order to directly test this hypothesis, Mgat1-/- blastocysts were positively identified by polymerase chain reaction (PCR) of genomic DNA. Reverse transcriptase PCR (RT-PCR) of RNA isolated from the same blastocysts, and restriction analysis of the PCR products, revealed that Mgat1 null blastocysts contained Mgat1 RNA derived from the wild-type Mgat1 gene. Consistent with this, all 3.5 day blastocysts from five heterozygous crosses bound the lectin L-PHA, a lectin previously shown not to bind to E8.5 or E9.5 Mgat1-/- embryos that lack complex N-glycans (Ioffe and Stanley (1994) Proc. Natl. Acad. Sci., USA, 91, 728-732). Blastocysts of 4.5 days postcoitum (dpc) obtained by culturing 3.5 dpc blastocysts also bound L-PHA. However, mutant embryos that did not bind L-PHA were present among progeny from E5.5 onward. Therefore, the effects of the Mgat1 null mutation are not operative until sometime between implantation and E5.5, due to the continued presence of maternally derived Mgat1 mRNA in preimplantation embryos.


Clinical drug-drug interactions of bosentan, a potent endothelial receptor antagonist, with various drugs: Physiological role of enzymes and transporters.
Bosentan, an endothelin-1 (ET) receptor antagonist is an important drug for the effective management of patients with pulmonary arterial hypertension. Bosentan has a rather complicated pharmacokinetics in humans involving multiple physiological components that have a profound influence on its drug disposition. Bosentan is mainly metabolized by cytochrome P450 (CYP) 3A4 and 2C9 enzymes with the involvement of multiple transporters that control its hepatic uptake and biliary excretion. The involvement of phase 2 metabolism of bosentan is a key to have an enhanced biliary excretion of the drug-related products. While bosentan exhibits high protein binding restricting the drug from extensive distribution and significant urinary excretion, bosentan induces its own metabolism by an increased expression of CYP3A4 on repeated dosing. Due to the above properties, bosentan has the potential to display drug-drug interaction with the co-administered drugs, either being a perpetrator or a victim. The intent of this review is manifold: a) to summarize the physiological role of CYP enzymes and hepatic-biliary transporters; b) to discuss the mechanism(s) involved in the purported liver injury caused by bosentan; c) to tabulate the numerous clinical drug-drug interaction studies involving the physiological interplay with CYP and/or transporters; d) to provide some perspectives on dosing strategy of bosentan.


Applications of modern ferroelectrics.
Long viewed as a topic in classical physics, ferroelectricity can be described by a quantum mechanical ab initio theory. Thin-film nanoscale device structures integrated onto Si chips have made inroads into the semiconductor industry. Recent prototype applications include ultrafast switching, cheap room-temperature magnetic-field detectors, piezoelectric nanotubes for microfluidic systems, electrocaloric coolers for computers, phased-array radar, and three-dimensional trenched capacitors for dynamic random access memories. Terabit-per-square-inch ferroelectric arrays of lead zirconate titanate have been reported on Pt nanowire interconnects and nanorings with 5-nanometer diameters. Finally, electron emission from ferroelectrics yields cheap, high-power microwave devices and miniature x-ray and neutron sources.


Hydrogen-rich saline protects against hepatic injury induced by ischemia-reperfusion and laparoscopic hepatectomy in swine.
Hydrogen-rich saline (HRS) has antioxidative, anti-inflammatory and anti-apoptotic properties. We investigated the effects of hydrogen on hepatic ischemia-reperfusion (I/R) and laparoscopic hepatectomy in swine. Twenty-one healthy Bama miniature pigs were randomly divided into the sham group, ischemia-reperfusion injury (IRI) group, HRS-5 (5 mL/kg) group, and HRS-10 (10 mL/kg) group. HRS was injected through the portal vein 10 min before reperfusion and at postoperative day 1, 2 and 3. The roles of HRS on oxidative stress, inflammatory response and liver regeneration were studied. Compared with the IRI group, HRS treatment attenuated oxidative stress by increasing catalase activity and reducing myeloperoxidase. White blood cells in the HRS-10 group were reduced compared with the IRI group (P < 0.01). In the HRS-10 group, interleukin-1 beta, interleukin-6 and tumor necrosis factor alpha, C-reactive protein and cortisol were downregulated, whereas interleukin-10 was upregulated. In addition, HRS attenuated endothelial cell injury and promoted the secretion of angiogenic cytokines, including vascular endothelial growth factor, angiopoietin-1 and angiopoietin-2. HRS elevated the levels of hepatocyte growth factor, Cyclin D1, proliferating cell nuclear antigen, Ki-67 and reduced the secretion of transforming growth factor-beta. HRS treatment may exert a protective effect against I/R and hepatectomy-induced hepatic damage by reducing oxidative stress, suppressing the inflammatory response and promoting liver regeneration.


Gauss-Legendre quadrature method used to evaluate the spatio-temporal intensity of ultrashort pulses in the focal region of lenses.
We analyze the spatio-temporal intensity of sub-20 femtosecond pulses with a carrier wavelength of 810 nm along the optical axis of low numerical aperture achromatic and apochromatic doublets designed in the IR region by using the scalar diffraction theory. The diffraction integral is solved by expanding the wave number around the carrier frequency of the pulse in a Taylor series up to third order, and then the integral over the frequencies is solved by using the Gauss-Legendre quadrature method. The numerical errors in this method are negligible by taking 96 nodes and the computational time is reduced by 95% compared to the integration method by rectangles. We will show that the third-order group velocity dispersion (GVD) is not negligible for 10 fs pulses at 810 nm propagating through the low numerical aperture doublets, and its effect is more important than the propagation time difference (PTD). This last effect, however, is also significant. For sub-20 femtosecond pulses, these two effects make the use of a pulse shaper necessary to correct for second and higher-order GVD terms and also the use of apochromatic optics to correct the PTD effect. The design of an apochromatic doublet is presented in this paper and the spatio-temporal intensity of the pulse at the focal region of this doublet is compared to that given by the achromatic doublet.


Society of Breast Imaging residency and fellowship training curriculum.
A recently developed Society of Breast Imaging curriculum for residency training is intended to provide guidance to residents and their mentors, and to practicing radiologists who want to keep up to date in screening, diagnosis, and interventional procedures. The curriculum contains lists of key concepts in 14 subject areas: epidemiology; anatomy; pathology, and physiology; equipment and technique; quality control; interpretation; problem-solving mammography; ultrasound; interventional procedures; reporting and medicolegal aspects; screening; MR imaging; therapeutic considerations; and patient management principles. The curriculum also makes recommendations about residency training, including the number of examinations the resident should interpret, and the time the resident should spend in breast imaging. Recommendations for fellowship training are also discussed.


Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex.
Systematic interrogation of gene function requires the ability to perturb gene expression in a robust and generalizable manner. Here we describe structure-guided engineering of a CRISPR-Cas9 complex to mediate efficient transcriptional activation at endogenous genomic loci. We used these engineered Cas9 activation complexes to investigate single-guide RNA (sgRNA) targeting rules for effective transcriptional activation, to demonstrate multiplexed activation of ten genes simultaneously, and to upregulate long intergenic non-coding RNA (lincRNA) transcripts. We also synthesized a library consisting of 70,290 guides targeting all human RefSeq coding isoforms to screen for genes that, upon activation, confer resistance to a BRAF inhibitor. The top hits included genes previously shown to be able to confer resistance, and novel candidates were validated using individual sgRNA and complementary DNA overexpression. A gene expression signature based on the top screening hits correlated with markers of BRAF inhibitor resistance in cell lines and patient-derived samples. These results collectively demonstrate the potential of Cas9-based activators as a powerful genetic perturbation technology.


Disparities in Psychiatric Diagnosis and Treatment for Youth with Systemic Lupus Erythematosus: Analysis of a National US Medicaid Sample.
To estimate the national prevalence and racial/ethnic differences in psychiatric diagnoses and pharmacologic treatment in a US Medicaid beneficiary population of youth with systemic lupus erythematosus (SLE). We included youth aged 10 to 18 years with a diagnosis of SLE (defined as ≥ 3 outpatient visit claims with an International Classification of Diseases, 9th ed. code of 710.0, each > 30 days apart) in the US Medicaid Analytic Extract database from 2006 and 2007. This database contains all inpatient and outpatient Medicaid claims from 49 states and the District of Columbia. We calculated the prevalence of psychiatric diagnoses and treatment, and used logistic regression to compare depression and anxiety diagnoses, antidepressant, and anxiolytic use among racial/ethnic groups. Of 970 youth with SLE, 15% were white, 42% were African American, 27% were Latino, and 16% were of other races/ethnicities. Diagnoses of depression were present for 19%, anxiety for 7%, acute stress/adjustment for 6%, and other psychiatric disorders for 18%. Twenty percent were prescribed antidepressants, 7% were prescribed anxiolytics, 6% were prescribed antipsychotics, and 5% were prescribed stimulants. In adjusted analyses, African Americans were less likely than whites to be diagnosed with depression (OR 0.56, 95% CI 0.34-0.90) or anxiety (OR 0.49, 95% CI 0.25-0.98), or to be prescribed anxiolytics (OR 0.23, 95% CI 0.11-0.48). We present population-level estimates showing high psychiatric morbidity in youth with SLE, but less prevalent diagnosis and treatment in African Americans. Mental health interventions should address potential racial/ethnic disparities in care.


Electrode-free picoinjection of microfluidic drops.
Existing methods for injecting reagents into drops utilize metal electrodes integrated into the microfluidic chip, adding cumbersome and error-prone steps to the fabrication process. Here, we introduce a method that uses the injection fluid itself as an electrode by exploiting dissolved electrolytes in the solution. This obviates the need for metal electrodes and reduces fabrication time and complexity while also affording precise control of the injected reagent volumes.


Clinical review: Special populations--critical illness and pregnancy.
Critical illness is an uncommon but potentially devastating complication of pregnancy. The majority of pregnancy-related critical care admissions occur postpartum. Antenatally, the pregnant patient is more likely to be admitted with diseases non-specific to pregnancy, such as pneumonia. Pregnancy-specific diseases resulting in ICU admission include obstetric hemorrhage, pre-eclampsia/eclampsia, HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome, amniotic fluid embolus syndrome, acute fatty liver of pregnancy, and peripartum cardiomyopathy. Alternatively, critical illness may result from pregnancy-induced worsening of pre-existing diseases (for example, valvular heart disease, myasthenia gravis, and kidney disease). Pregnancy can also predispose women to diseases seen in the non-pregnant population, such as acute respiratory distress syndrome (for example, pneumonia and aspiration), sepsis (for example, chorioamnionitis and pyelonephritis) or pulmonary embolism. The pregnant patient may also develop conditions co-incidental to pregnancy such as trauma or appendicitis. Hemorrhage, particularly postpartum, and hypertensive disorders of pregnancy remain the most frequent indications for ICU admission. This review focuses on pregnancy-specific causes of critical illness. Management of the critically ill mother poses special challenges. The physiologic changes in pregnancy and the presence of a second, dependent, patient may necessitate adjustments to therapeutic and supportive strategies. The fetus is generally robust despite maternal illness, and therapeutically what is good for the mother is generally good for the fetus. For pregnancy-induced critical illnesses, delivery of the fetus helps resolve the disease process. Prognosis following pregnancy-related critical illness is generally better than for age-matched non-pregnant critically ill patients.


Differential effects of the anticonvulsant topiramate on neurobehavioral and histological outcomes following traumatic brain injury in rats.
The efficacy of topiramate, a novel therapeutic agent approved for the treatment of seizure disorders, was evaluated in a model of traumatic brain injury (TBI). Adult male rats were anesthetized (sodium pentobarbital, 60 mg/kg, i.p.), subjected to lateral fluid percussion brain injury (n = 60) or sham injury (n = 47) and randomized to receive either topiramate or vehicle at 30 min (30 mg/kg, i.p.), and 8, 20 and 32 h postinjury (30 mg/kg, p.o.). In Study A, memory was evaluated using a Morris water maze at 48 h postinjury, after which brain tissue was evaluated for regional cerebral edema. In Study B, animals were evaluated for motor function at 48 h and 1, 2, 3, and 4 weeks postinjury using a composite neuroscore and the rotating pole test and for learning ability at 4 weeks. Brains were analyzed for hemispheric tissue loss and hippocampal CA3 cell loss. Topiramate had no effect on posttraumatic cerebral edema or histologic damage when compared to vehicle. At 48 h, topiramate treatment improved memory function in sham but not brain-injured animals, while at one month postinjury it impaired learning performance in brain-injured but not sham animals. Topiramate significantly improved composite neuroscores at 4 weeks postinjury and rotating pole performance at 1 and 4 weeks postinjury, suggesting a potentially beneficial effect on motor function following TBI.


The complexity of crime network data: a case study of its consequences for crime control and the study of networks.
The field of social network analysis has received increasing attention during the past decades and has been used to tackle a variety of research questions, from prevention of sexually transmitted diseases to humanitarian relief operations. In particular, social network analyses are becoming an important component in studies of criminal networks and in criminal intelligence analysis. At the same time, intelligence analyses and assessments have become a vital component of modern approaches in policing, with policy implications for crime prevention, especially in the fight against organized crime. In this study, we have a unique opportunity to examine one specific Swedish street gang with three different datasets. These datasets are the most common information sources in studies of criminal networks: intelligence, surveillance and co-offending data. We use the data sources to build networks, and compare them by computing distance, centrality, and clustering measures. This study shows the complexity factor by which different data sources about the same object of study have a fundamental impact on the results. The same individuals have different importance ranking depending on the dataset and measure. Consequently, the data source plays a vital role in grasping the complexity of the phenomenon under study. Researchers, policy makers, and practitioners should therefore pay greater attention to the biases affecting the sources of the analysis, and be cautious when drawing conclusions based on intelligence assessments and limited network data. This study contributes to strengthening social network analysis as a reliable tool for understanding and analyzing criminality and criminal networks.


Polyunsaturated Fatty Acid EAB-277® Supplementation Improved Heart Rate Variability and Clinical Signs in Tracheal Collapse Dogs.
Canine tracheal collapse is a progressive disease in small breed dogs resulting from chronic inflammation of the tracheal mucosal lining. Polyunsaturated fatty acid EAB-277® is one of the nutraceuticals that can alleviate inflammation and oxidative stress. Heart rate variability (HRV) is a prognostic tool related to sympathovagal balance and oxidative stress level, which is widely used with cardiorespiratory diseases. However, the effect of EAB-277® on HRV in tracheal collapse dogs has rarely been investigated. In this study, 26 tracheal collapse dogs were divided into two groups. In the control group, the dogs received the standard treatment, whereas the dogs in the EAB-277® group received standard treatment combined with EAB-277®. After being treated for 5 weeks, changes in radiographic findings, blood profiles, serum malondialdehyde, inflammatory markers, and HRV were evaluated. This study found that clinical signs were improved in both groups (p < 0.05). However, serum malondialdehyde (MDA), Interleukin-6 (IL-6), and Tumor necrosis factor-alpha (TNF-α) were decreased only in the EAB-277® group after treatment for five weeks (p < 0.05) and the mean percent change of MDA, IL-6, and TNF-α at week five compared to baseline in the EAB-277® group was greater than in the control group (p < 0.05). Additionally, greater sympathovagal imbalance indicated by decreased standard deviation of all normal R-R intervals (SDNN) and standard deviation of the averaged R-R intervals for all 5-minutes segments (SDANN) was found in the control group at week five compared to baseline (P < 0.05), whereas EAB-277® improved SDNN and SDANN and decreased low frequency/high-frequency component (LF/HF ratio) after being treated for five weeks (P < 0.05). This study demonstrates that EAB-277® improves clinical signs and attenuates HRV impairment by reducing oxidative stress and inflammation in tracheal collapse dogs.


Importance of acid-base equilibrium in electrocatalytic oxidation of formic acid on platinum.
Electro-oxidation of formic acid on Pt in acid is one of the most fundamental model reactions in electrocatalysis. However, its reaction mechanism is still a matter of strong debate. Two different mechanisms, bridge-bonded adsorbed formate mechanism and direct HCOOH oxidation mechanism, have been proposed by assuming a priori that formic acid is the major reactant. Through systematic examination of the reaction over a wide pH range (0-12) by cyclic voltammetry and surface-enhanced infrared spectroscopy, we show that the formate ion is the major reactant over the whole pH range examined, even in strong acid. The performance of the reaction is maximal at a pH close to the pKa of formic acid. The experimental results are reasonably explained by a new mechanism in which formate ion is directly oxidized via a weakly adsorbed formate precursor. The reaction serves as a generic example illustrating the importance of pH variation in catalytic proton-coupled electron-transfer reactions.


Relationships between age and epi-genotype of the FMR1 exon 1/intron 1 boundary are consistent with non-random X-chromosome inactivation in FM individuals, with the selection for the unmethylated state being most significant between birth and puberty.
Methylation of the fragile X-related epigenetic element 2 (FREE2) located on the exon 1/intron 1 boundary of the FMR1 gene is related to FMRP expression and cognitive impairment in full mutation (FM; CGG>200) individuals. We examined the relationship between age, the size of the FMR1 CGG expansion and the methylation output ratio (MOR) at 12 CpG sites proximal to the exon 1/intron 1 boundary using FREE2 MALDI-TOF MS. The patient cohort included 119 males and 368 females, i.e. 121 healthy controls (CGG<40), 176 premutation (CGG 55-170) and 190 FM (CGG 213-2000). For all CpG units examined, FM males showed a significantly elevated MOR compared with that in hypermethylated FM females. In FM males the MOR for most CpG units significantly positively correlated with both age and CGG size (P< 0.05). In FM females the skewing towards the unmethylated state was significant for half of the units between birth and puberty (P < 0.05). The methylation status of intron 1 CpG10-12 that was most significantly related to cognitive impairment in our earlier study, did not change significantly with age in FM females. These results challenge the concept of fragile X syndrome (FXS)-related methylation being static over time, and suggest that due to the preference for the unmethylated state in FM females, X-inactivation at this locus is not random. The findings also highlight that the prognostic value of FXS methylation testing is not uniform between all CpG sites, and thus may need to be evaluated on a site-by-site basis.


One ligand, two regulators and three binding sites: How KDPG controls primary carbon metabolism in Pseudomonas.
Effective regulation of primary carbon metabolism is critically important for bacteria to successfully adapt to different environments. We have identified an uncharacterised transcriptional regulator; RccR, that controls this process in response to carbon source availability. Disruption of rccR in the plant-associated microbe Pseudomonas fluorescens inhibits growth in defined media, and compromises its ability to colonise the wheat rhizosphere. Structurally, RccR is almost identical to the Entner-Doudoroff (ED) pathway regulator HexR, and both proteins are controlled by the same ED-intermediate; 2-keto-3-deoxy-6-phosphogluconate (KDPG). Despite these similarities, HexR and RccR control entirely different aspects of primary metabolism, with RccR regulating pyruvate metabolism (aceEF), the glyoxylate shunt (aceA, glcB, pntAA) and gluconeogenesis (pckA, gap). RccR displays complex and unusual regulatory behaviour; switching repression between the pyruvate metabolism and glyoxylate shunt/gluconeogenesis loci depending on the available carbon source. This regulatory complexity is enabled by two distinct pseudo-palindromic binding sites, differing only in the length of their linker regions, with KDPG binding increasing affinity for the 28 bp aceA binding site but decreasing affinity for the 15 bp aceE site. Thus, RccR is able to simultaneously suppress and activate gene expression in response to carbon source availability. Together, the RccR and HexR regulators enable the rapid coordination of multiple aspects of primary carbon metabolism, in response to levels of a single key intermediate.


Aqueous Huaier Extract Suppresses Gastric Cancer Metastasis and Epithelial to Mesenchymal Transition by Targeting Twist.
Trametes robiniophila Murr. (Huaier) is a widely used anti-cancer agent in China. Strong evidence for the anti-proliferative activity of Huaier has been reported; however, its anti-metastatic potential against gastric cancer (GC) as well as its underlying mechanism of action are unknown. Here, we show that treatment with an aqueous Huaier extract over a range of concentrations significantly suppressed both the invasiveness and migratory ability of GC cells. Huaier could also partly reverse the epithelial-mesenchymal transition (EMT), as characterized by increased expression of the epithelial marker E-cadherin and decreased expression of the mesenchymal markers N-cadherin and vimentin. In addition, Huaier-treated cells expressed lower levels of Twist compared to untreated controls, and overexpression of Twist via transfection could partially abolish the anti-metastatic activity of Huaier. Furthermore, elevated Twist expression was correlated with an advanced TNM stage, a high rate of lymph node metastasis, and reduced disease-free survival in GC patients. These findings reveal a novel anti-metastatic mechanism for Huaier, which inhibits the EMT by targeting Twist, suggesting its potential application against a GC relapse.


Interaction of Mycobacterium avium complex with human macrophages: roles of membrane receptors and serum proteins.
Invasive, disease-associated members of the Mycobacterium avium complex are facultative intracellular pathogens of mammalian macrophages. The mechanism(s) by which M. avium is ingested by mononuclear phagocytes is unknown. We examined the role of membrane receptors on macrophages as well as the role of opsonic components of the serum (complement, fibronectin, C-reactive protein and fibrinogen in the attachment and ingestion of M. avium by human monocyte-derived macrophages. Preincubation of serum with antibodies against C3 and fibronectin, in contrast to preincubation of serum with antibodies against complement-reactive protein and fibrinogen, significantly reduced the binding of M. avium to macrophages in concentration-dependent manner (57 to 93% and 35 to 61% inhibition by anti-C3 and anti-fibronectin antibody, respectively, in a concentration range of 25 to 100 micrograms/ml). We also observed that incubation of macrophages with OKM1 anti-complement receptor type 3 (CR3) antibody in the presence of serum decreased the binding of M. avium to macrophages by 86% +/- 6%, while the same antibody inhibited binding by 63% +/- 7% in the absence of serum. In contrast, incubation of macrophages with anti-LFA-1, anti-p 150.95, anti-CR1, or anti-Mac-1 had no effect on the ability of M. avium to bind to the cell. In addition, incubation of macrophages with alpha-methyl-D-mannoside was also associated with decreased attachment of M. avium to macrophages. These results provide evidence for the role of three macrophage receptors (CR3, fibronectin, and mannosyl-fucosyl receptors) in the uptake of M. avium by human macrophages.


Ccdc94 protects cells from ionizing radiation by inhibiting the expression of p53.
DNA double-strand breaks (DSBs) represent one of the most deleterious forms of DNA damage to a cell. In cancer therapy, induction of cell death by DNA DSBs by ionizing radiation (IR) and certain chemotherapies is thought to mediate the successful elimination of cancer cells. However, cancer cells often evolve to evade the cytotoxicity induced by DNA DSBs, thereby forming the basis for treatment resistance. As such, a better understanding of the DSB DNA damage response (DSB-DDR) pathway will facilitate the design of more effective strategies to overcome chemo- and radioresistance. To identify novel mechanisms that protect cells from the cytotoxic effects of DNA DSBs, we performed a forward genetic screen in zebrafish for recessive mutations that enhance the IR-induced apoptotic response. Here, we describe radiosensitizing mutation 7 (rs7), which causes a severe sensitivity of zebrafish embryonic neurons to IR-induced apoptosis and is required for the proper development of the central nervous system. The rs7 mutation disrupts the coding sequence of ccdc94, a highly conserved gene that has no previous links to the DSB-DDR pathway. We demonstrate that Ccdc94 is a functional member of the Prp19 complex and that genetic knockdown of core members of this complex causes increased sensitivity to IR-induced apoptosis. We further show that Ccdc94 and the Prp19 complex protect cells from IR-induced apoptosis by repressing the expression of p53 mRNA. In summary, we have identified a new gene regulating a dosage-sensitive response to DNA DSBs during embryonic development. Future studies in human cancer cells will determine whether pharmacological inactivation of CCDC94 reduces the threshold of the cancer cell apoptotic response.


Epidemiology of Methicillin-Resistant Staphylococcus aureus Diabetic Foot Infections in a Large Academic Hospital: Implications for Antimicrobial Stewardship.
Diabetic foot infections (DFIs) are the leading cause of non-traumatic lower extremity amputations in the United States. Antimicrobials active against methicillin-resistant Staphylococcus aureus (MRSA) are recommended in patients with associated risk factors; however, limited data exist to support these recommendations. Due to the changing epidemiology of MRSA, and the consequences of unnecessary antibiotic therapy, guidance regarding the necessity of empirical MRSA coverage in DFIs is needed. We sought to 1) describe the prevalence of MRSA DFIs at our institution and compare to the proportion of patients who receive MRSA antibiotic coverage and 2) identify risk factors for MRSA DFI. This was a retrospective cohort study of all adult, culture-positive DFI patients managed at University Hospital, San Antonio, TX between January 1, 2010 and September 1, 2014. Patient eligibility included a principal ICD-9-CM discharge diagnosis code for foot infection and a secondary diagnosis of diabetes. The primary outcome was MRSA identified in the wound culture. Independent variables assessed included patient demographics, comorbidities, prior hospitalization, DFI therapies, prior antibiotics, prior MRSA infection, and laboratory values. Multivariable logistic regression was used to identify risk factors for MRSA DFI. Overall, 318 patients met inclusion criteria. Patients were predominantly Hispanic (79%) and male (69%). Common comorbidities included hypertension (76%), dyslipidemia (52%), and obesity (49%). S. aureus was present in 46% of culture-positive DFIs (MRSA, 15%). A total of 273 patients (86%) received MRSA antibiotic coverage, resulting in 71% unnecessary use. Male gender (OR 3.09, 95% CI 1.37-7.99) and bone involvement (OR 1.93, 1.00-3.78) were found to be independent risk factors for MRSA DFI. Although MRSA was the causative pathogen in a small number of DFI, antibiotic coverage targeted against MRSA was unnecessarily high.


Defining phenotypic and functional heterogeneity of glioblastoma stem cells by mass cytometry.
Most patients with glioblastoma (GBM) die within 2 years. A major therapeutic goal is to target GBM stem cells (GSCs), a subpopulation of cells that contribute to treatment resistance and recurrence. Since their discovery in 2003, GSCs have been isolated using single-surface markers, such as CD15, CD44, CD133, and α6 integrin. It remains unknown how these single-surface marker-defined GSC populations compare with each other in terms of signaling and function and whether expression of different combinations of these markers is associated with different functional capacity. Using mass cytometry and fresh operating room specimens, we found 15 distinct GSC subpopulations in patients, and they differed in their MEK/ERK, WNT, and AKT pathway activation status. Once in culture, some subpopulations were lost and previously undetectable ones materialized. GSCs that highly expressed all 4 surface markers had the greatest self-renewal capacity, WNT inhibitor sensitivity, and in vivo tumorigenicity. This work highlights the potential signaling and phenotypic diversity of GSCs. Larger patient sample sizes and antibody panels are required to confirm these findings.


Effects of inbred/outbred crosses on progeny sex ratio in Silene latifolia (Caryophyllaceae).
Sex ratio polymorphism has been extensively studied in Silene latifolia, but it is neither known whether inbreeding (which is likely to occur under field conditions) affects it, nor which of the proposed mechanisms (Y degeneration, X-linked drive) is more important. Both mechanisms predict reduced pollen performance. In this study, females were crossed with pollen from related and unrelated males in single-donor and two-donor crosses, and the sex ratio of offspring (n = 866, 60 crosses), sons'in vitro pollen germination and sex ratios in parental families were scored. Flowers receiving only unrelated pollen produced a significant excess of sons. Sex ratios were not significantly correlated between generations. Sons'in vitro pollen germination was significantly negatively correlated with the 'sex-ratio phenotype' of maternal grandfathers, but not of fathers. This generation leap may be consistent with X-linked determinants because Y-linked determinants alone cannot explain it (grandfathers, fathers and sons share the same Y chromosome). Further work is required, but inbreeding and limited dispersal may lead to local accumulation of biasing factors, a process potentially countered by conditional shifts to produce more sons in pure outbred crosses.


Cyclophosphamide dose intensification during induction therapy for intermediate-risk pediatric rhabdomyosarcoma is feasible but does not improve outcome: a report from the soft tissue sarcoma committee of the children's oncology group.
More than half of pediatric rhabdomyosarcoma cases have intermediate-risk features and suboptimal outcome (3-year failure-free survival estimates, 55 to 76%). Dose intensification of known active agents may improve outcome. This pilot study evaluated the feasibility of dose intensification of cyclophosphamide in previously untreated patients ages < 21 years with intermediate-risk rhabdomyosarcoma. Induction therapy comprised four 3-week cycles of VAC: vincristine (V) 1.5 mg/m2 on days 0, 7, and 14; actinomycin D (A) 1.35 mg/m2 on day 0; and dose-intensified cyclophosphamide (C) on days 0, 1, and 2. The three cyclophosphamide dose levels tested were as follows: (a) 1.2 g/m2/dose; (b) 1.5 g/m2/dose; and (c) 1.8 g/m2/dose. Continuation therapy comprised nine additional cycles of VAC with 2.2 g/m2/cycle of C. Radiotherapy was administered at week 0 (parameningeal tumors with intracranial extension) or week 12 or 15 (all others). Between October 1996 and August 1999, 115 eligible patients were enrolled. Three of 15 patients treated at dose level 2 experienced life-threatening dose-limiting toxicity (typhlitis +/- other severe toxicity). Dose level 1 was the maximum-tolerated dose, and 91 evaluable patients were treated at this level. The 3-year failure-free and overall survival estimates for patients treated at the maximum-tolerated dose were 52% (95% confidence interval, 41-64%) and 67% (95% confidence interval, 56-77%), respectively, at a median follow-up of 3 years. A 64% increase in the standard cyclophosphamide dosage during induction (to 3.6 g/m2/cycle) was tolerated. However, outcomes were similar to those observed at lower dosages, suggesting that alkylator dose intensification does not benefit patients with intermediate-risk rhabdomyosarcoma.


Excitable properties in astrocytes derived from human embryonic CNS stem cells.
Although it is widely believed that astrocytes lack excitability in adult tissue, primitive action potential-like responses have been elicited from holding potentials negative to -80 mV, in cultured and injury-induced gliotic rodent astrocytes and in human glia under pathological conditions such as glioblastomas and temporal lobe epilepsy. The present study was designed to investigate the properties of astrocytes (identified by immunoreactivity for glial fibrillary acidic protein) derived from multipotent human embryonic CNS stem cells and cultured for 12-25 days in differentiating conditions. We describe here for the first time that brief (1 ms) current pulses elicit spikes from a resting potential (VREST) of approximately -37 mV and, more interestingly, that spontaneous firing can be occasionally recorded in human astrocytes. A voltage-clamp study revealed that in these cells: (i) the half-inactivation of the tetrodotoxin (TTX)-sensitive Na+ channels is around VREST; (ii) the delayed rectifier K+ current is very small; (iii) the ever-present transient outward A-type K+ channels are paradoxically capable of inhibiting the action potentials elicited from a negative membrane potential (-55 to -60 mV); and (iv) inwardly rectifying currents are not present. The responses predicted from a simulation model are in agreement with the experiments. As suggested by recent studies, the decrease of Na+ channel expression and the changes of the electrophysiological properties during the postnatal maturation of the CNS seem to exclude the possibility that astrocytes may play an excitable role in adult tissue. Our data show that excitability and firing should be considered an intrinsic attribute of human astrocytes during CNS development. This is likely to have physiological importance because the role of astrocytes during development is different from the [K+]o-buffering role played in adult CNS, namely the glutamate release and/or the guiding of migrating neurons.


Priority setting for the prevention and control of cardiovascular diseases: multi-criteria decision analysis in four eastern Mediterranean countries.
To explore the feasibility of using a simple multi-criteria decision analysis method with policy makers/key stakeholders to prioritize cardiovascular disease (CVD) policies in four Mediterranean countries: Palestine, Syria, Tunisia and Turkey. A simple multi-criteria decision analysis (MCDA) method was piloted. A mixed methods study was used to identify a preliminary list of policy options in each country. These policies were rated by different policymakers/stakeholders against pre-identified criteria to generate a priority score for each policy and then rank the policies. Twenty-five different policies were rated in the four countries to create a country-specific list of CVD prevention and control policies. The response rate was 100% in each country. The top policies were mostly population level interventions and health systems' level policies. Successful collaboration between policy makers/stakeholders and researchers was established in this small pilot study. MCDA appeared to be feasible and effective. Future applications should aim to engage a larger, representative sample of policy makers, especially from outside the health sector. Weighting the selected criteria might also be assessed.


Designing focused chemical libraries enriched in protein-protein interaction inhibitors using machine-learning methods.
Protein-protein interactions (PPIs) may represent one of the next major classes of therapeutic targets. So far, only a minute fraction of the estimated 650,000 PPIs that comprise the human interactome are known with a tiny number of complexes being drugged. Such intricate biological systems cannot be cost-efficiently tackled using conventional high-throughput screening methods. Rather, time has come for designing new strategies that will maximize the chance for hit identification through a rationalization of the PPI inhibitor chemical space and the design of PPI-focused compound libraries (global or target-specific). Here, we train machine-learning-based models, mainly decision trees, using a dataset of known PPI inhibitors and of regular drugs in order to determine a global physico-chemical profile for putative PPI inhibitors. This statistical analysis unravels two important molecular descriptors for PPI inhibitors characterizing specific molecular shapes and the presence of a privileged number of aromatic bonds. The best model has been transposed into a computer program, PPI-HitProfiler, that can output from any drug-like compound collection a focused chemical library enriched in putative PPI inhibitors. Our PPI inhibitor profiler is challenged on the experimental screening results of 11 different PPIs among which the p53/MDM2 interaction screened within our own CDithem platform, that in addition to the validation of our concept led to the identification of 4 novel p53/MDM2 inhibitors. Collectively, our tool shows a robust behavior on the 11 experimental datasets by correctly profiling 70% of the experimentally identified hits while removing 52% of the inactive compounds from the initial compound collections. We strongly believe that this new tool can be used as a global PPI inhibitor profiler prior to screening assays to reduce the size of the compound collections to be experimentally screened while keeping most of the true PPI inhibitors. PPI-HitProfiler is freely available on request from our CDithem platform website, www.CDithem.com.


The management of isolated distal radius fractures in children.
In a consecutive series of 175 forearm fractures requiring manipulation in children who presented to the John Radcliffe Hospital between 1 January 1991 and 30 June 1992, 23 had isolated distal radius fractures, all of which required manipulation for unacceptable angulation or displacement. We had previously noted that these fractures had a high incidence of loss of position leading to malunion. The 23 patients were randomly allocated to one of two treatment groups: either manipulation and cast alone, or manipulation and percutaneous Kirschner wiring with cast. These two groups were followed up clinically and radiographically until union occurred. Those fractures treated by percutaneous wiring had no significant complications, and all had a satisfactory result. The fractures treated by manipulation and casting alone required further manipulation in 10 cases (91%). We conclude that these problematic fractures should be treated by stabilisation with percutaneous wiring.


Neural probe combining microelectrodes and a droplet-based microdialysis collection system for high temporal resolution sampling.
We propose a novel neural probe which combines microfluidic channels with recording and stimulation electrodes. The developed microfabrication approach enables the concentration of every active element such as electrodes and the sampling inlet in close proximity on the same surface. As a first approach, full functional validation is presented in this work (in vivo testing will be presented in the next study). Electrical characterization by impedance spectroscopy is performed in order to assess the electrode properties. An advanced experimental setup enabling the validation of the fluidic functions of the neural probe is also presented. It allowed the achievement of a high temporal resolution (170 ms) during sampling as a result of the integration of a T-junction droplet generator inside the probe. The droplets reached a volume of 0.84 nL and are separated by a non-aqueous phase (perfluoromethyldecalin, PFD). This probe represents an innovative tool for neuroscientists as it can be implanted in precise brain structures while combining electrical stimulation with sampling at a high temporal resolution.


Data communication between brain implants and computer.
Recent advances in neuroscience, microelectronics, and information technology have allowed construction of miniature, but highly intelligent, devices to be implanted within the brain to perform in vitro diagnostic and therapeutic functions. However, there exists a significant problem in establishing an effective wireless data communication link between brain implants and external computer. This communication investigates this link and presents a new design using the mechanism of volume conduction of biological tissues. A theoretical model of volume conduction of the head is utilized to compute signal strength in data communication and the result is evaluated by a physical model. The two-way data communication sensitivity of the volume conduction channel is found to be symmetric, as suggested by the reciprocity theorem. A high-performance, x-shaped volume conduction antenna has been designed. Experiments are performed on animals which demonstrate the effectiveness of this volume conduction approach.


Identification of Subtypes of Barrett's Esophagus and Esophageal Adenocarcinoma Based on DNA Methylation Profiles and Integration of Transcriptome and Genome Data.
Esophageal adenocarcinomas (EACs) are heterogeneous and often preceded by Barrett's esophagus (BE). Many genomic changes have been associated with development of BE and EAC, but little is known about epigenetic alterations. We performed epigenetic analyses of BE and EAC tissues and combined these data with transcriptome and genomic data to identify mechanisms that control gene expression and genome integrity. In a retrospective cohort study, we collected tissue samples and clinical data from 150 BE and 285 EAC cases from the Oesophageal Cancer Classification and Molecular Stratification consortium in the United Kingdom. We analyzed methylation profiles of all BE and EAC tissues and assigned them to subgroups using non-negative matrix factorization with k-means clustering. Data from whole-genome sequencing and transcriptome studies were then incorporated; we performed integrative methylation and RNA-sequencing analyses to identify genes that were suppressed with increased methylation in promoter regions. Levels of different immune cell types were computed using single-sample gene set enrichment methods. We derived 8 organoids from 8 EAC tissues and tested their sensitivity to different drugs. BE and EAC samples shared genome-wide methylation features, compared with normal tissues (esophageal, gastric, and duodenum; controls) from the same patients and grouped into 4 subtypes. Subtype 1 was characterized by DNA hypermethylation with a high mutation burden and multiple mutations in genes in cell cycle and receptor tyrosine signaling pathways. Subtype 2 was characterized by a gene expression pattern associated with metabolic processes (ATP synthesis and fatty acid oxidation) and lack methylation at specific binding sites for transcription factors; 83% of samples of this subtype were BE and 17% were EAC. The third subtype did not have changes in methylation pattern, compared with control tissue, but had a gene expression pattern that indicated immune cell infiltration; this tumor type was associated with the shortest time of patient survival. The fourth subtype was characterized by DNA hypomethylation associated with structure rearrangements, copy number alterations, with preferential amplification of CCNE1 (cells with this gene amplification have been reported to be sensitive to CDK2 inhibitors). Organoids with reduced levels of MGMT and CHFR expression were sensitive to temozolomide and taxane drugs. In a comprehensive integrated analysis of methylation, transcriptome, and genome profiles of more than 400 BE and EAC tissues, along with clinical data, we identified 4 subtypes that were associated with patient outcomes and potential responses to therapy.


Coffee, Caffeine, and Health Outcomes: An Umbrella Review.
To evaluate the associations between coffee and caffeine consumption and various health outcomes, we performed an umbrella review of the evidence from meta-analyses of observational studies and randomized controlled trials (RCTs). Of the 59 unique outcomes examined in the selected 112 meta-analyses of observational studies, coffee was associated with a probable decreased risk of breast, colorectal, colon, endometrial, and prostate cancers; cardiovascular disease and mortality; Parkinson's disease; and type-2 diabetes. Of the 14 unique outcomes examined in the 20 selected meta-analyses of observational studies, caffeine was associated with a probable decreased risk of Parkinson's disease and type-2 diabetes and an increased risk of pregnancy loss. Of the 12 unique acute outcomes examined in the selected 9 meta-analyses of RCTs, coffee was associated with a rise in serum lipids, but this result was affected by significant heterogeneity, and caffeine was associated with a rise in blood pressure. Given the spectrum of conditions studied and the robustness of many of the results, these findings indicate that coffee can be part of a healthful diet.


Influence of hippophae rhamnoides on two appetite factors, gastric emptying and metabolic parameters, in children with functional dyspepsia.
Our aim was to explore in children with functional dyspepsia the effect of hippophae rhamnoides on the levels of plasma appetite factors and on their gastrointestinal motility. A hundred and twenty children with functional dyspepsia were randomly divided into three groups: Group I (treated with hippophae rhamnoides), Group II (treated with domperidone), and Group III (treated with hippophae rhamnoides plus domperidone). The treatment lasted for eight weeks. The levels of plasma leptin (LP) and neuropeptide Y (NPY) were measured before and after treatment. All patients underwent a gastric emptying (GE) test by ultrasound (US) to measure the rate of postprandial gastric antrum residual, at 30min, 60min, 90min and 120min. The average value of subcutaneous fat, body fat percentage, upper arm girth and body mass index (BMI) were also measured. To compare the US with the radionuclide GE test 14 healthy adults volunteers were tested by both GE techniques. We found that the levels of LP and NPY in plasma were markedly higher after treatment in Groups I and III than in Group II. The postprandial gastric antrum remains at 60min, 90min and 120min in Groups I and III fell greatly and the thickness of skin fold (SF), body fat percentage and arm girth increased (P<0.05). The GE half emptying time of a mixed liquid-solid food measured by B US and by the radionuclide technique in the same individuals was similar (P>0.05). In conclusion, in children's functional dyspepsia, our study showed that hippophae rhamnoides increases the levels of appetite factors, leptin and neuropeptide Y, increases gastric emptying and gastrointestinal digestive function, children's growth and development.


Acetylcholine receptors from human muscle as pharmacological targets for ALS therapy.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease affecting motor neurons that leads to progressive paralysis of skeletal muscle. Studies of ALS have revealed defects in expression of acetylcholine receptors (AChRs) in skeletal muscle that occur even in the absence of motor neuron anomalies. The endocannabinoid palmitoylethanolamide (PEA) modified the clinical conditions in one ALS patient, improving muscle force and respiratory efficacy. By microtransplanting muscle membranes from selected ALS patients into Xenopus oocytes, we show that PEA reduces the desensitization of acetylcholine-evoked currents after repetitive neurotransmitter application (i.e., rundown). The same effect was observed using muscle samples from denervated (non-ALS) control patients. The expression of human recombinant α1β1γδ (γ-AChRs) and α1β1εδ AChRs (ε-AChRs) in Xenopus oocytes revealed that PEA selectively affected the rundown of ACh currents in ε-AChRs. A clear up-regulation of the α1 subunit in muscle from ALS patients compared with that from non-ALS patients was found by quantitative PCR, but no differential expression was found for other subunits. Clinically, ALS patients treated with PEA showed a lower decrease in their forced vital capacity (FVC) over time as compared with untreated ALS patients, suggesting that PEA can enhance pulmonary function in ALS. In the present work, data were collected from a cohort of 76 ALS patients and 17 denervated patients. Our results strengthen the evidence for the role of skeletal muscle in ALS pathogenesis and pave the way for the development of new drugs to hamper the clinical effects of the disease.


The first house officer year: views of graduate and non-graduate entrants to medical school.
To determine whether graduate and non-graduate entrants to medical school differ in their views on the first year spent in medical practice as a pre-registration house officer. We carried out postal questionnaire surveys of medical qualifiers of 1999, 2000 and 2002 from all UK medical schools, 1 year after qualification. The timing of the study slightly pre-dates the recent major expansion in graduate entry fast-track courses. Differences between graduate and non-graduate entrants were few and, even when statistically significant, were small in scale. Graduate entrants viewed their working hours, pay and living conditions at work, such as hospital accommodation and food, a little less favourably than did non-graduate entrants. Graduate entrants were also less satisfied than non-graduates with time available for family, social and recreational activities. However, graduate entrants were more likely than non-graduate entrants to feel positive about their future career prospects. There were no differences between graduate and non-graduate entrants in whether they felt they had been well prepared by their medical schools for the jobs they undertook as house officers. Levels of job satisfaction expressed by graduate and non-graduate entrants were similar, as were their responses to most other statements about attitudes to clinical work. 'Quality of life' issues, a sense of being fairly rewarded, and expectations about one's physical working environment seem a little more important to graduate than to non-graduate entrants. Apart from these, the findings suggest that graduate status, at entry to medical school, has no appreciable influence on attitudes to the work of a junior hospital doctor.


A scientific and animal welfare assessment of the OECD Health Effects Test Guidelines for the safety testing of chemicals under the European Union REACH system.
We have assessed each of the OECD Health Effects Test Guidelines (TGs) that were included in an annex to the Internet consultation issued by the European Commission relating to the Registration, Evaluation and Authorisation of Chemicals (REACH) legislation for the testing of new and existing chemical substances. Each guideline has been analysed with respect to its design and its scientific and animal welfare implications, the extent to which it makes use of modern techniques, and its suitability to be used in the REACH system for the testing of large numbers of chemicals. The scientific basis of the test and its justification are considered, as well as the numbers of animals required, and the potential adverse effects on them. The prospects and possibilities for applying the Three Rs (reduction, refinement and replacement) to each of the TGs are also discussed. We have proposed an overall testing strategy for how these TGs and other methods could best be deployed for chemicals testing, should it be necessary to fill data gaps. Certain TGs have been omitted from the strategy, when we have considered them to be unnecessary for chemicals testing. A series of recommendations has been made for improving the TGs with regard to both their scientific content and ways in which they could be better designed in relation to optimising the use of the animals concerned, and minimising adverse welfare consequences to them. Our investigations show that there is an urgent need to update the TGs to reflect modern techniques and methods, and to use current approaches for applying refinement strategies to improve the scientific and animal welfare aspects of the procedures used. Improvements can and should be made in all aspects of toxicity testing, from sample preparation, and animal housing, care and feeding, to dose formulation, test material administration, and the histopathological and clinical analysis of tissue samples. Opportunities for streamlining individual assays are very limited, but testing could be made more efficient by: a) only undertaking studies that provide relevant data; b) making greater use of screens and preliminary testing; c) applying some tests simultaneously to the same animals; d) using one sex; and e) eliminating redundant tests. In conclusion, it is clear that, as they stand, the OECD Health Effects TGs are unsuitable for use in the European Union REACH system, for which potentially very large numbers of laboratory animals will be needed for the testing of a very large number of chemicals.


A fish model for the study of the relationship between neuroendocrine and immune cells in the intestinal epithelium: Silurus glanis infected with a tapeworm.
Immunohistochemical, immunofluorescence and ultrastructural studies were conducted on a sub-population of 20 wels catfish Silurus glanis from a tributary of the River Po (Northern Italy). Fish were examined for the presence of ecto- and endo-parasites; in the intestine of 5 fish, 11 specimens of cestode Glanitaenia osculata were noted and was the only helminth species encountered. The architecture of intestine and its cellular features were nearly identical in either the uninfected S. glanis or in those harboring G. osculata. Near the site of worm's attachment, mucous cells, several mast cells (MCs), few neutrophils and some endocrine cells (ECs) were found to co-occur within the intestinal epithelium. MCs and neutrophils were abundant also in the submucosa. Immunohistochemical staining revealed that enteric ECs were immunoreactive to met-enkephalin, galanin and serotonin anti-bodies. The numbers of ECs, mucous cells and MCs were significantly higher in infected wels catfish (Mann-Whitney U test, p < 0.05). Dual immunofluorescence staining with the biotinylated lectin Sambucus nigra Agglutinin and the rabbit polyclonal anti-met-enkephalin or anti-serotonin, with parallel transmission electron microscopy, showed that ECs often made intimate contact with the mucous cells and epithelial MCs. The presence of numerous MCs in intestinal epithelium shows S. glanis to be an interesting model fish to study processes underlying intestinal inflammation elicited by an enteric worm. Immune cells, ECs and mucous cells of the intestinal epithelium have been described at the ultrastructural level and their possible functions and interactions together will be discussed.


Hepatic JAK2 protects against atherosclerosis through circulating IGF-1.
Atherosclerosis is considered both a metabolic and inflammatory disease; however, the specific tissue and signaling molecules that instigate and propagate this disease remain unclear. The liver is a central site of inflammation and lipid metabolism that is critical for atherosclerosis, and JAK2 is a key mediator of inflammation and, more recently, of hepatic lipid metabolism. However, precise effects of hepatic Jak2 on atherosclerosis remain unknown. We show here that hepatic Jak2 deficiency in atherosclerosis-prone mouse models exhibited accelerated atherosclerosis with increased plaque macrophages and decreased plaque smooth muscle cell content. JAK2's essential role in growth hormone signalling in liver that resulted in reduced IGF-1 with hepatic Jak2 deficiency played a causal role in exacerbating atherosclerosis. As such, restoring IGF-1 either pharmacologically or genetically attenuated atherosclerotic burden. Together, our data show hepatic Jak2 to play a protective role in atherogenesis through actions mediated by circulating IGF-1 and, to our knowledge, provide a novel liver-centric mechanism in atheroprotection.


Calibration of an integrating sphere for determining the absorption coefficient of scattering suspensions.
Measuring the absolute absorption of suspensions of absorbing particles with unknown scattering characteristics is not possible in conventional spectrophotometers or in integrating spheres that have the sample located outside the sphere. A method for the calibration and use of an integrating sphere with a centrally located sample to measure absolute absorption coefficients of scattering suspensions is presented. Under the tested conditions the integrating sphere used in this study was insensitive to changes in the scattering coefficient of the sample but had a nonlinear response to increasing absorption of the sample, which could be corrected with an empirically derived function. This response was analyzed by using a Monte Carlo simulation, and results indicated that amplification of the absorption signal was primarily due to photons reflected from the sphere surface and the baffle reentering the cuvette. The calibration procedure described here may be generally applicable to spheres of different configurati n. An example of the use of the sphere for determining the absorption and scattering coefficients of marine phytoplankton samples is presented.


Intimate partner violence and preschoolers' explicit memory functioning.
This research examines whether parents' intimate partner physical violence (IPV) relates to their preschoolers' explicit memory functioning, whether children's symptoms of hyperarousal mediate this relation, and whether mothers' positive parenting moderates this relation. Participants were 69 mothers and their 4- or 5-year-old child (34 girls). Mothers completed measures of IPV, children's hyperarousal symptoms, parent-child aggression, and positive parenting. Measures of explicit memory functioning were administered to preschoolers. As expected, IPV correlated negatively with preschoolers' performance on explicit memory tasks, even after controlling for parent-child aggression and demographic variables related to preschoolers' memory functioning. Preschoolers' hyperarousal symptoms did not mediate the relation between IPV and explicit memory functioning, but mothers' positive parenting moderated this relation. Specifically, the negative relation between IPV and preschoolers' performance on 2 of the 3 explicit memory tasks was weaker when mothers engaged in higher levels of positive parenting. These findings extend research on IPV and children's adjustment difficulties to explicit memory functioning in preschoolers and suggest that mothers can ameliorate the influence of IPV on preschoolers' memory functioning via their parenting.


Modulation of vinblastine resistance in metastatic renal cell carcinoma with cyclosporine A or tamoxifen: a cancer and leukemia group B study.
Multidrug resistance mediated by P-glycoprotein may be an important cause of chemotherapy failure. Renal cell carcinoma is a disease known to demonstrate a high degree of intrinsic chemotherapy drug resistance, and this has been shown to be related to intrinsic overexpression of P-glycoprotein. Cyclosporine A and tamoxifen have been shown to reverse multidrug resistance in renal cell carcinoma cell lines in vitro. Phase I studies have defined appropriate doses of cyclosporine A and tamoxifen that can be combined with continuous-infusion vinblastine and safely achieve serum levels associated in vitro with resistance reversal. A randomized Phase II study was carried out by the Cancer and Leukemia Group B to evaluate the potential of high doses of cyclosporine A or tamoxifen to modulate clinical vinblastine resistance in patients with advanced renal cell carcinoma. Patients were treated initially with continuous-infusion vinblastine alone (1.2 mg/m2/day for 4 days or 1.5 mg/m2/day for 5 days); patients with stable or progressive disease were then treated with the same vinblastine regimen, combined with a high-dose regimen of either cyclosporine A (12.5 mg/kg/day for 5 days) or tamoxifen (400 mg/m2 as a loading dose and 300 mg/m2/day for 13 days). Sixty-three patients were randomized to each arm. Eighty patients on both arms were evaluable for response to vinblastine alone; of these, only one patient achieved a partial response. Thirty-three patients went on to be treated with vinblastine and high-dose cyclosporine A. No responses were observed, although four patients with progressive disease on prior vinblastine achieved stabilization of disease after cyclosporine A was added. Addition of cyclosporine resulted in more leukopenia (5% versus 25%) and in transient hyperbilirubinemia (24%) and neurocortical changes (11%). No significant azotemia was observed. Thirty-five patients received high-dose tamoxifen with continuous-infusion vinblastine. One complete remission was seen in a patient who had stable disease only with prior vinblastine alone; no other responses were observed. Leukopenia was not more severe with the addition of tamoxifen to vinblastine, nor was hyperbilirubinemia observed. However, 9% of patients developed transient ataxia with or without neurocortical changes as a result of high-dose tamoxifen therapy, and 11% developed phlebitis. We conclude that advanced renal cell carcinoma is a highly chemoresistant tumor, that continuous-infusion vinblastine has no appreciable activity in the therapy of this disease, and that addition of high doses of cyclosporine A or tamoxifen was not able to modulate this resistance in these patients. Suggestions regarding study design for future drug resistance modulation trials were made based on the design and conduct of this study.


Trends in antenatal care attendance and health facility delivery following community and health facility systems strengthening interventions in Northern Uganda.
Maternal morbidity and mortality remains high in Uganda; largely due to inadequate antenatal care (ANC), low skilled deliveries and poor quality of other maternal health services. In order to address both the demand and quality of ANC and skilled deliveries, we introduced community mobilization and health facility capacity strengthening interventions. Interventions were introduced between January 2010 and September 2011. These included: training health workers, provision of medical supplies, community mobilization using village health teams, music dance and drama groups and male partner access clubs. These activities were implemented at Kitgum Matidi health center III and its catchment area. Routinely collected health facility data on selected outcomes in the year preceding the interventions and after 21 months of implementation of the interventions was reviewed. Trend analysis was performed using excel and statistical significance testing was performed using EPINFO StatCal option. The number of pregnant women attending the first ANC visit significantly increased from 114 to 150 in the first and fourth quarter of 2010 (OR 1.72; 95% CI 1.39-2.12) and to 202 in the third quarter of 2011(OR 11.41; 95% CI 7.97-16.34). The number of pregnant women counselled, tested and given results for HIV during the first ANC attendance significantly rose from 92 (80.7%) to 146 (97.3%) in the first and fourth quarter of 2010 and then to 201 (99.5%) in the third quarter of 2011. The number of male partners counseled, tested and given results together with their wives at first ANC visit rose from 13 (16.7%) in the fourth quarter of 2009 to 130 (89%) in the fourth quarter of 2010 and to 180 (89.6%) in the third quarter of 2011. There was a significant rise in the number of pregnant women delivering in the health facility with provision of mama-kits (delivery kits), from 74 (55.2%) to 149 (99.3%) in the second and fourth quarter of 2010. Combined community and facility systems strengthening interventions led to increased first ANC visits by women and their partners, and health facility deliveries. Interventions aimed at increasing uptake of maternal health services should address both the demand and availability of quality services.


Using graph models to find transcription factor modules: the hitting set problem and an exact algorithm.
: Systematically perturbing a cellular system and monitoring the effects of the perturbations on gene expression provide a powerful approach to study signal transduction in gene expression systems. A critical step of revealing a signal transduction pathway regulating gene expression is to identify transcription factors transmitting signals in the system. In this paper, we address the task of identifying modules of cooperative transcription factors based on results derived from systems-biology experiments at two levels: First, a graph algorithm is developed to identify a minimum set of co-operative TFs that covers the differentially expressed genes under each systematic perturbation. Second, using a clique-finding approach, modules of TFs that tend to consistently cooperate together under various perturbations are further identified. Our results indicate that this approach is capable of identifying many known TF modules based on the individual experiment; thus we provide a novel graph-based method of identifying context-specific and highly reused TF-modules.