Altering CO binding on gold cluster cations by Pd-doping.
The introduction of dopant atoms into metal nanoparticles is an effective way to control the interaction with adsorbate molecules and is important in many catalytic processes. In this work, experimental and theoretical evidence of the influence of Pd doping on the bonding between small cationic AuN+ clusters and CO is presented. The CO adsorption is studied by combining low-pressure collision cell reactivity and infrared multiple photon dissociation spectroscopy experiments with density functional theory calculations. Measured dissociation rates of cluster-CO complexes (N ≤ 21) allow the estimation of cluster-CO binding energies, showing that Pd doping increases the CO adsorption energy to an extent that is size-dependent. These trends are reproduced by theoretical calculations up to N = 13. In agreement with theory, measurements of the C-O vibrational frequency suggest that for the doped PdAuN-1+ (N = 3-5, 11) clusters, CO adsorbs on an Au atom, while for N = 6-10 and N = 12-14, CO interacts directly with the Pd dopant. A pronounced red-shifting of the C-O vibrational frequency is observed when CO interacts directly with the Pd dopant, indicating a significant back-donation of electron charge from Pd to CO. In contrast, the blue-shifted frequencies, observed when CO interacts with an Au atom, indicate that σ-donation dominates the Au-CO interaction. Studying such systems at the sub-nanometre scale enables a fundamental comprehension of the interactions between adsorbates, dopants and the host (Au) species at the atomic level.


Preservatives in cosmetics in the Israeli market conform well to the EU legislation.
Preservatives are important and frequent skin sensitizers, found in a wide range of products for personal and occupational use. According to the European legislation, some cosmetic ingredients are restricted in terms of quantity and a detailed list of ingredients must be present on the product or packaging. To examine the use of preservatives in common cosmetics on the Israeli market. Sixty different Israeli brand cosmetics, including shampoos, liquid soaps, body creams and hand creams were randomly selected. Ingredient labels were examined. The products were investigated by the chromotropic acid method for release of formaldehyde and by high performance liquid chromatography for the presence of formaldehyde, DMDM hydantoin and methylchloroisothiazolinone/methylisothiazolinone (MCI/MI) and MI content. All products but one contained a detailed list of ingredients printed on the package. According to labelling, the most prevalent preservatives in Israeli shampoos and liquid soaps were DMDM hydantoin and MCI/MI. Hand creams and body creams contained mainly parabens but also iodopropynyl butylcarbamate, phenoxyethanol and DMDM hydantoin. Formaldehyde in doses from 4 to 429 ppm, and DMDM hydantoin were detected in 38 and 16 (63% and 27%) of the products, respectively. MCI/MI was detected in 11 (18%) of the products, with highest prevalence in rinse- off products (55%). Excluding one hand cream which measured 106 ppm MI, the amount of formaldehyde, DMDM hydantoin, MCI/MI and MI was within the allowed concentrations by the European directive in all cases. In Israel, adaptation of the European directive prevails, as shown by the measurements we performed on randomly selected products.


A Pediatric Burn Outpatient Short Stay Program Decreases Patient Length of Stay With Equivalent Burn Outcomes.
Traditionally, small pediatric burns are managed with inpatient admission and daily dressing changes. In 2011, our burn center implemented an outpatient short stay (OSS) program in which small pediatric burns were managed as an outpatient utilizing Mepilex AgTM dressings changed under moderate sedation every 5 to 7 days. Pediatric burn cases were queried for 2 time periods: before the OSS program (2009-2010) and after the OSS program (2013-2014). Burns > 15% total body surface area (TBSA), children with polytrauma, and children > 10 years old were excluded. Independent t tests and chi-square tests were conducted to analyze differences in patient demographics, burn management, and burn outcomes between these groups. Two hundred nineteen cases were included in the analysis (77 pre-OSS and 142 post-OSS). There was no difference in patient age (P = 0.872) or TBSA (P = 0.786) between the groups. The post-OSS group had shorter inpatient length of stay (2.93 days vs 5.21 days; P < 0.001) and fewer dressing changes (2.32 vs 4.71; P < 0.001). There were no changes in readmission rates (P = 0.375) or burns requiring grafting (P = 0.155). Although not reaching statistical significance, less children in the post-OSS group had infectious complications (P = 0.054) or required reoperation in a 2-year follow-up period (P = 0.081). Patient and family satisfaction with the program was high. Children treated after the implementation of an OSS burn program at the University of Wisconsin had decreased inpatient length of stay and fewer painful burn dressing changes. These patients exhibited equivalent, if not superior burn outcomes.


New insight into the immunomodulatory mechanisms of Tretinoin in NMRI mice.
Recent evidence have proposed that Tretinoin produced in the gut preferentially promote differentiation of FoxP3+Treg cells but inhibits Th17 lymphocytes, and this may be the main immunomdulatory mechanism of Tretinoin in vivo. This study was done to investigate the effects of Tretinoin in outbred white mice after challenge with sheep red blood cells (SRBC). Twenty male NMRI-mice randomly allocated in two equal groups. Mice were treated with 1×10(9) SRBCs emulsified in CFA intraperitoneally twice with one weak interval. Animals were bled 5 days after last injection. Moreover, 48 hr before bleeding time, 1×10(9) SRBCs were injected into the left hind foot pad of mice. Tretinoin (25 mg/kg-every other day) were intraperitoneally injected into the treatment group from the beginning of the study and continued throughout the study. The levels of anti-SRBC antibody and the specific cellular immune responses were measured by microhemagglutination test and footpad thickness, respectively. Moreover, splenocytes were checked for proliferation rate, respiratory burst, cytokine production and FoxP3+Treg cells frequency. Tretinoin markedly alleviated cellular immunity and concurrently potentiated humoral immunity after mice challenge with SRBCs. Furthermore, aside from reducing NBT reduction and lymphocyte proliferation, Tretinoin markedly suppressed the secretion of interleukin-17 and conversely, increased the production of interleukin-10. However, the level of IFN-γ and the frequency of FoxP3+Treg cells did not alter significantly. The in vivo immunomudlatoty effects of Tretinoin may be partly due to immune deviation from pro-inflammatory cytokine interleukin-17 to anti-inflammatory cytokine interleukin-10, but not absolutely depend on the expansion of FoxP3(+)Treg cells.


Itch and motivation to scratch: an investigation of the central and peripheral correlates of allergen- and histamine-induced itch in humans.
Intense itch and urge to scratch are the major symptoms of many chronic skin ailments, which are increasingly common. Vicious itch-scratch cycles are readily established and may diminish quality of life for those afflicted. We investigated peripheral and central processing of two types of itch sensation elicited by skin-prick tests of histamine and allergen solutions. Itch-related skin blood flow changes were measured by laser Doppler in 14 subjects responsive to type I allergens and 14 nonatopic subjects. In addition, this study examined central processing of both types of itch using functional magnetic resonance imaging (fMRI). Itch perception and blood flow changes were significantly greater when itch was induced by allergens compared with histamine. Both types of itch correlated significantly with activity in the genual anterior cingulate, striatum, and thalamus. Moreover, itch elicited by allergens activated orbitofrontal, supplementary motor, and posterior parietal areas. Histamine-induced itch also significantly correlated with activation in the insula bilaterally. The identification of limbic and ventral prefrontal activation in two types of itch processing likely reflects the subjects' desire to relieve the itch sensation by scratching, and these regions have been repeatedly associated with motivation processing. A dysfunction of the striato-thalamo-orbitofrontal circuit is believed to underlie the failure to regulate motivational drive in disorders associated with strong urges, e.g., addiction and obsessive compulsive disorder. The patterns of itch-induced activation reported here may help explain why chronic itch sufferers frequently self-harm through uncontrollable itch-scratch cycles.


Hydrogen Bond versus Halogen Bond in Cation-Cation Complexes: Effect of the Solvent.
Competition between hydrogen- (HB) and halogen-bonded (XB) 4-ammoniumpyridine and halogenammonium (NHn F3-n X+ ; n=0-3; X=F, Cl, Br, and I) cation-cation complexes are explored by means of DFT calculations. HB and XB minima structures are found for all systems in the gas phase. As the number of fluorine atoms increases, the HB complexes are more favored than those of XB. Proton transfer is generally observed in complexes with two, three, or four halogen atoms. The XB complexes evolve from traditional halogen bonds, to halogen-shared complexes, and to ionic complexes as the number of fluorine atoms increases. The dissociation transition states and their corresponding barriers are also characterized; the barriers increase as the number of fluorine atoms increases. The results if solvent effects are considered indicate that, even in an apolar solvent, such as n-hexane, most of the complexes have favorable binding energies. Atoms-in-molecules theory is used to analyze the complexes, and results in good correlations between electron density and total electron energy density (Η) values with the intermolecular bond length. According to the Η values obtained, the covalency of these interactions starts to manifest at distances around 72-74 % the sum of the van der Waals radii of the interacting atoms.


Hazardous Drug Exposure: Case Report Analysis From a Prospective, Multisite Study of Oncology Nurses' Exposure in Ambulatory Settings.
Hazardous drug exposure is an occupational health hazard to oncology nurses. Sparse data are available regarding the frequency and characteristics of hazardous drug spills. This article aims to describe nurses' hazardous drug exposures and use of personal protective equipment during drug spills. The Drug Exposure Feedback and Education for Nurses' Safety study launched in March 2015. When drug spills occurred, consented RNs administering chemotherapy in ambulatory infusion settings completed brief questionnaires. Descriptive statistics were used to summarize equipment use and spill events. Spills were common, despite the use of closed-system transfer devices. Over two years, 51 nurses from 12 participating academic infusion centers reported 61 unique spills. Spills commonly involved highly toxic drugs. Personal protective equipment use during drug spills was suboptimal. These foundational data reveal gaps in clinical practice.


MO-A-BRB-02: Facts and Fiction of Flattening Filter Free (FF-FFF) X-Rays Beams.
The primary purpose of the FFF X-rays is to provide much higher dose rates available for treatments. For example, FFF X-rays from Varian TrueBEAM can deliver 1400 MU/minute for 6 MV X-rays and 2400 MU/minutes for 10 MV X-rays. Higher dose rates have definite clinical benefits in organ motion management. For example, larger dose fractions can be delivered in a single breath-hold or gated portion of a breathing cycle. In SRS or SBRT treatments, large MUs are often required and FFF X-ray beams can deliver these large MUs in much shorter "beam-on" time. With shorten treatment time, these FFF X-rays improve patient comfort and dose delivery accuracy. FFF X-ray beams may become one of the necessary equipment configurations for SBRT and/or SRS treatments, in the future. This presentation will address some unique issues dealing with the FFF X-rays: (1) FIELD SIZES flattening Filter Free (FFF) X-ray beam has been in clinical use for quite some time. However, not until recently, these FFF beams are used in limited, small field sizes, for example, in Tomotherapy and CyberKnife machines.Varian TrueBEAM allows the FFF X-ray beam to have up to 40 × 40 cm field sizes for both 6 and 10 MV X-rays (15 MV FFF X-rays are not yet released for clinical use). For large treatment fields, the dose uniformity within an irradiated treatment field will need to be "modulated" by MLC movements (IMRT) to cut down the higher beam intensity near the central portion of the FFF X-ray beam. Thus, larger MUs are required compared with a conventional (flattened) X-ray beam. Or, MLC movements (EVIRT) are now being used to "flatten" the FFF X-rays to provide dose uniformity within those large PTVs. The high dose rates from the FFF X-rays are now being off-set by the larger MUs requirements. Therefore, FFF X-rays can bring clinical advantages over conventional X-rays when used with small field sizes, such as in SBRT and/or SRS applications. (2) DOSEVIETRY MEASUREMENT EQUIMENT: Because of the more than 2 to 4 fold increases in dose rate (MU/minute), the radiation measurement equipment and techniques need to be carefully evaluated, such as ionization chamber characteristics, electrometers, scanning equipment. First comes to mind is the ion-recombination characteristics of the ionization chamber (P-ion). This will determine the accuracies of the measured percentage depth doses and penumbra of these FFF X-ray fields. And, it will also affect the absolute dose measurements (Gy/MU) using the TG-51 formulations. The measured PDDs and profiles should be corrected for the P-ion effect. However, it is not a simple task for physicists to perform the P-ion corrections for PDDs and profiles using the presently available methods associated in commercial 3-D scanning equipment and algorithm. It may become necessary for physicists to adapt and get accustom to the use of "standard beam data" provided by manufacturers in the future. In addition, because of the use of FFF X-rays are focused on SBRT and/or SRS applications, beam data acquisition, scanning techniques, and beam modeling are vitally important. There are many publications addressing the "output factors" from small fields, but none pay enough attention to the penumbra characteristics of these small X-ray beams. Because of the proximity to critical organs, the penumbra characteristics of small fields are often more clinically important than output factors. FFF X-rays play an important role in SBRT and SRS applications. Therefore, careful penumbra measurements should be addressed. Again, it may become necessary for physicists to adapt the use of "standard beam data" provided by manufacturers. (3) RADIO-BIOLOGICAL QUESTION: Though there is a lack of controlled clinical studies with FFF X-ray beams, there are several scientific articles addressing the radiobiological concerns of high dose rate deliveries, especially when it is used to deliver large doses per fraction, such as 10 Gy/faction. This type of dose per fraction is often used in SBRT or SRS treatments. Radiobiological concerns are not in the cell kill effect within the target volume. It is the normal tissue damages surrounding the target. There are concerns about the late toxicities of these high dose rate and high dose per fraction deliveries using FFF X-ray beams. (4) SKIN (ENTRANCE) DOSE DISCUSSIONS: In conventional X-ray beams, the low energy components of the X-ray beam are removed by the in-line X-ray flattening filter. But, in Flatten Filter Free X-ray beams, these low energy components are exiting the X-ray collimators. This is clearly documented by the difference in the percentage depth doses for these FFF X-ray beams. The FFF X-ray beams have a lower "effective energy" compared to conventional X-rays. Therefore, it is important to study the skin (entrance) dose from these FFF X-rays. In the literature, reported skin (entrance) doses from different linear accelerator manufacturers vary widely. Skin doses from Varian TrueBEAM have been studied and have found to be marginally higher than the conventional X-rays. However this margin increase is not clinically significant.(5) Summary / conclusions / discussions: The FFF X-rays improve the treatment delivery by their very high dose rates (1400 and 2400 MU/minute) and shortened treatment time. FFF X-ray beams are most applicable and the high dose rates are most advantageous when the treatment field sizes are small. The dosimetry of FFF-X-rays is made more complex by the P-ion determination and necessary corrections to X-ray beam percentage depth doses and profiles. There are radiobiological concerns about late toxicity of normal tissue irradiated by FFF X-rays when large dose per faction treatment applications are used. There are wide ranges of skin doses from these FFF X-rays reported in the literature. 1. Flattening Filter Free X-rays have been in clinical use for many years but mostly for small field sizes. 2. Flattening Filter Free X-rays have significantly shorter treatment time if is used for small field applications, such as: SBRT or SRS. 3. Because of the high dose rate (MU/minute), dosimetric properties of these FFF X-rays need to be carefully studied. Ion-recombination of ionization chambers is a concern. Beam data acquisition, beam modeling, and absolute dosimetry need to be done with great care, especially in small field applications. 4. Late toxicities of normal tissue may be a concern and need to be studied by organized clinical protocols.


Comparison of the Myxobolus fauna of common barbel from Hungary and Iberian barbel from Portugal.
We compared Myxobolus infection of common barbel Barbus barbus from the Danube River in Hungary with that in Iberian barbel Luciobarbus bocagei from the Este River in Portugal. In Hungary, we recorded 5 known Myxobolus species (M. branchialis, M. caudatus, M. musculi, M. squamae, and M. tauricus) and described M. branchilateralis sp. n. In Portugal we recorded 6 Myxobolus species (M. branchialis, M. branchilateralis sp. n., M. cutanei, M. musculi, M. pfeifferi, and M. tauricus). Species found in the 2 habitats had similar spore morphology and only slight differences were observed in spore shape or measurements. All species showed a specific tissue tropism and had a definite site selection. M. branchialis was recorded from the lamellae of the gills, large plasmodia of M. branchilateralis sp. n. developed at both sides of hemibranchia, M. squamae infected the scales, plasmodia of M. caudatus infected the scales and the fins, and M. tauricus were found in the fins and pin bones. In the muscle, 3 species, M. musculi, M. pfeifferi and M. tauricus were found; however they were found in distinct locations. Plasmodia of M. musculi developed intracellularly in muscle cells, plasmodia of M. tauricus were found in the dense connective tissue of the pin bones, whereas M. pfeifferi formed plasmodia in the connective tissue of the intramuscular septa. This latter species was often found in the cartilaginous gill arch as well. Comparative morphological and phylogenetic studies, as well as 18S rDNA sequences, revealed differences between the Myxobolus fauna of the 2 barbel species originating from different geographic regions.


Crystal structures and kinetics of monofunctional proline dehydrogenase provide insight into substrate recognition and conformational changes associated with flavin reduction and product release.
Proline dehydrogenase (PRODH) catalyzes the FAD-dependent oxidation of proline to Δ(1)-pyrroline-5-carboxylate, which is the first step of proline catabolism. Here, we report the structures of proline dehydrogenase from Deinococcus radiodurans in the oxidized state complexed with the proline analogue L-tetrahydrofuroic acid and in the reduced state with the proline site vacant. The analogue binds against the si face of the FAD isoalloxazine and is protected from bulk solvent by helix α8 and the β1-α1 loop. The FAD ribityl chain adopts two conformations in the E-S complex, which is unprecedented for flavoenzymes. One of the conformations is novel for the PRODH superfamily and may contribute to the low substrate affinity of Deinococcus PRODH. Reduction of the crystalline enzyme-inhibitor complex causes profound structural changes, including 20° butterfly bending of the isoalloxazine, crankshaft rotation of the ribityl, shifting of α8 by 1.7 Å, reconfiguration of the β1-α1 loop, and rupture of the Arg291-Glu64 ion pair. These changes dramatically open the active site to facilitate product release and allow electron acceptors access to the reduced flavin. The structures suggest that the ion pair, which is conserved in the PRODH superfamily, functions as the active site gate. Mutagenesis of Glu64 to Ala decreases the catalytic efficiency 27-fold, which demonstrates the importance of the gate. Mutation of Gly63 decreases the efficiency 140-fold, which suggests that flexibility of the β1-α1 loop is essential for optimal catalysis. The large conformational changes that are required to form the E-S complex suggest that conformational selection plays a role in substrate recognition.


The skull of Homo naledi.
The species Homo naledi was recently named from specimens recovered from the Dinaledi Chamber of the Rising Star cave system in South Africa. This large skeletal sample lacks associated faunal material and currently does not have a known chronological context. In this paper, we present comprehensive descriptions and metric comparisons of the recovered cranial and mandibular material. We describe 41 elements attributed to Dinaledi Hominin (DH1-DH5) individuals and paratype U.W. 101-377, and 32 additional cranial fragments. The H. naledi material was compared to Plio-Pleistocene fossil hominins using qualitative and quantitative analyses including over 100 linear measurements and ratios. We find that the Dinaledi cranial sample represents an anatomically homogeneous population that expands the range of morphological variation attributable to the genus Homo. Despite a relatively small cranial capacity that is within the range of australopiths and a few specimens of early Homo, H. naledi shares cranial characters with species across the genus Homo, including Homo habilis, Homo rudolfensis, Homo erectus, and Middle Pleistocene Homo. These include aspects of cranial form, facial morphology, and mandibular anatomy. However, the skull of H. naledi is readily distinguishable from existing species of Homo in both qualitative and quantitative assessments. Since H. naledi is currently undated, we discuss the evolutionary implications of its cranial morphology in a range of chronological frameworks. Finally, we designate a sixth Dinaledi Hominin (DH6) individual based on a juvenile mandible.


Phase-Contrast Radiography Enables Detection of Early Changes in Articular Cartilage in a Mouse Model of Osteoarthritis.
The aim of this study was to investigate the potential of using phase-contrast radiography to detect early changes in articular cartilage in a mouse model of osteoarthritis. An iodoacetate-induced mouse model of osteoarthritis was used. Mice with iodoacetate-induced osteoarthritis were euthanized at 1 and 2 wks after injection. Specimens were obtained by sectioning mouse tissue into 5-mm-thick slices. Sections were examined using synchrotron x-rays from the 7B2 beamline at the Pohang Light Source. Magnified microradiographic images, obtained by phase-contrast radiography, were used to identify changes in articular cartilage and the subchondral bone. Refraction-enhanced phase-contrast radiography enabled detailed microscopic and macroscopic observation of articular cartilage by a noninvasive technique. Detailed microscopic images revealed partially damaged articular cartilage, containing an irregular trilaminar zone, at 1 wk after injection. At 2 wks after injection, a necrotic cleft was clearly visible, along with collapse of the trilaminar zones. Shrunken chondrocytes and chondrocyte clusters of variable sizes were also observed. Phase-contrast radiography is a feasible technique for the early detection of osteoarthritis and is capable of detecting microstructural changes in articular cartilage.


Subcellular distribution of copper in the kidneys of normal, copper-poisoned, and thiomolybdate-treated sheep.
Twenty-seven sheep given either copper (Cu) and/or tetrathiomolybdate (TM) were used to study the subcellular distribution of Cu within the kidney and to monitor the location of lysosomes within the subcellular fractions using acid phosphatase (AP) as a marker enzyme. Copper dosing alone increased the Cu content in the liver and the kidneys. The administration of intravenous TM prevented the development of chronic copper poisoning (CCP) in sheep, reduced the rate of accumulation of Cu in the liver of Cu-dosed animals, but increased the Cu content of kidneys in both the control and Cu-dosed sheep. The total amount of Cu that accumulated in the kidneys of sheep given TM appears to depend on several factors: a) liver Cu concentration, b) Cu intake, and c) dosage of TM. Thus, the highest Cu concentration was found in the kidneys of sheep that continued to receive Cu orally at the same time as they were given TM. The intracellular distribution of Cu and AP in the kidneys showed that in the control sheep given neither Cu or TM, the highest proportion of Cu was in the cytosol fraction, and the highest specific activity of AP was in the light mitochondrial (lysosomal) fraction. Dosing with Cu markedly increased the Cu concentration and greatly elevated the total activity of AP in the heavier fractions, i.e., the nuclear (N) and heavy mitochondrial (MH). Thus, the increase in Cu observed in the N and MH fractions was not caused by an accumulation of Cu by nuclei and mitochondria, but was due to an accumulation of Cu by lysosomes that sedimented with the heavier fractions. The intracellular distribution of Cu in the kidneys of TM-treated sheep was similar to that seen in Cu-loaded sheep. Although Cu accumulated readily in the kidneys of animals receiving TM, kidney function tests showed neither glomerular nor tubular functional impairment.


Stakeholder analysis for health research: case studies from low- and middle-income countries.
Future Health Systems: Innovations for Equity (FHS) is working in six partner countries in Asia and Africa, focusing on strengthening the research-policy interface in relation to specific health system research projects. These projects present an opportunity to study the influence of stakeholders on research and policy processes. Qualitative stakeholder analysis. Stakeholder analysis was conducted in each FHS country using a structured approach. A cross-country evaluation was performed concentrating on six key areas: chosen research topic; type of intervention considered; inclusion/exclusion of stakeholder groups; general stakeholder considerations; power level, power type and agreement level of stakeholders; and classification of and approaches to identified stakeholders. All six countries identified a range of stakeholders but each country had a different focus. Four of the six countries identified stakeholders in addition to the guidelines, while some of the stakeholder categories were not identified by countries. The mean power level of identified stakeholders was between 3.4 and 4.5 (1=very low; 5=very high). The percentage of classified stakeholders that were either drivers or supporters ranged from 60% to 91%. Three important common areas emerge when examining the execution of the FHS country stakeholder analyses: clarity on the purpose of the analyses; value of internal vs external analysts; and the role of primary vs secondary analyses. This paper adds to the global body of knowledge on the utilization of stakeholder analysis to strengthen the research-policy interface in the developing world.


Far-red fluorescent protein excitable with red lasers for flow cytometry and superresolution STED nanoscopy.
Far-red fluorescent proteins are required for deep-tissue and whole-animal imaging and multicolor labeling in the red wavelength range, as well as probes excitable with standard red lasers in flow cytometry and fluorescence microscopy. Rapidly evolving superresolution microscopy based on the stimulated emission depletion approach also demands genetically encoded monomeric probes to tag intracellular proteins at the molecular level. Based on the monomeric mKate variant, we have developed a far-red TagRFP657 protein with excitation/emission maxima at 611/657 nm. TagRFP657 has several advantages over existing monomeric far-red proteins including higher photostability, better pH stability, lower residual green fluorescence, and greater efficiency of excitation with red lasers. The red-shifted excitation and emission spectra, as compared to other far-red proteins, allows utilizing TagRFP657 in flow cytometry and fluorescence microscopy simultaneously with orange or near-red fluorescence proteins. TagRFP657 is shown to be an efficient protein tag for the superresolution fluorescence imaging using a commercially available stimulated emission depletion microscope.


Role of beta-adrenoceptor signaling in skeletal muscle: implications for muscle wasting and disease.
The importance of beta-adrenergic signaling in the heart has been well documented, but it is only more recently that we have begun to understand the importance of this signaling pathway in skeletal muscle. There is considerable evidence regarding the stimulation of the beta-adrenergic system with beta-adrenoceptor agonists (beta-agonists). Although traditionally used for treating bronchospasm, it became apparent that some beta-agonists could increase skeletal muscle mass and decrease body fat. These so-called "repartitioning effects" proved desirable for the livestock industry trying to improve feed efficiency and meat quality. Studying beta-agonist effects on skeletal muscle has identified potential therapeutic applications for muscle wasting conditions such as sarcopenia, cancer cachexia, denervation, and neuromuscular diseases, aiming to attenuate (or potentially reverse) the muscle wasting and associated muscle weakness, and to enhance muscle growth and repair after injury. Some undesirable cardiovascular side effects of beta-agonists have so far limited their therapeutic potential. This review describes the physiological significance of beta-adrenergic signaling in skeletal muscle and examines the effects of beta-agonists on skeletal muscle structure and function. In addition, we examine the proposed beneficial effects of beta-agonist administration on skeletal muscle along with some of the less desirable cardiovascular effects. Understanding beta-adrenergic signaling in skeletal muscle is important for identifying new therapeutic targets and identifying novel approaches to attenuate the muscle wasting concomitant with many diseases.


Can physical activity ameliorate immunosenescence and thereby reduce age-related multi-morbidity?
Remodelling of the immune system with age - immunosenescence - is a substantial contributor to poor health in older adults, with increasing risk of infections, cancer and chronic inflammatory disease contributing to age-related multi-morbidity. What is seldom considered when examining the immune response of an aged individual is that the immune system is profoundly influenced by physical activity. Habitual physical activity levels decline with age, with significant consequences for muscle mass and function. Skeletal muscle is a major immune regulatory organ and generates a range of proteins, termed myokines, which have anti-inflammatory and immunoprotective effects. Several studies indicate that maintaining physical activity has immune benefits in older adults, for example, it reduces the systemic inflammation associated with chronic age-related diseases. Here, we discuss how physical activity can prevent or ameliorate age-related multi-morbidity by boosting immune function, and we consider whether physical activity could improve immunotherapy outcomes in age-related conditions such as cancer.


Youth development program participation and intentional self-regulation skills: contextual and individual bases of pathways to positive youth development.
The present research used data from Grades 8, 9, and 10 of the 4-H Study of Positive Youth Development, a longitudinal study involving U.S. adolescents, in order to better elucidate the process through which the strengths of youth and the ecological resources promoting healthy development (such as out-of-school-time programs) may contribute to thriving. We examined the relationship between adolescents' self-regulation skills (selection, optimization, and compensation) and their participation in youth development (YD) programs across Grades 8 and 9 in predicting Grade 10 PYD and Contribution. Results indicated that while self-regulation skills alone predicted PYD, self regulation and YD program participation both predicted Contribution. In addition, Grade 8 YD participation positively predicted Grade 9 self regulation, which, in turn, predicted Grade 10 PYD and Contribution. We discuss how the alignment of youth strengths and resources within the environment may promote positive youth development.


Visceral adipose NLRP3 impairs cognition in obesity via IL-1R1 on CX3CR1+ cells.
Induction of the inflammasome protein cryopyrin (NLRP3) in visceral adipose tissue (VAT) promotes release of the proinflammatory cytokine IL-1β in obesity. Although this mechanism contributes to peripheral metabolic dysfunction, effects on the brain remain unexplored. We investigated whether visceral adipose NLRP3 impairs cognition by activating microglial IL-1 receptor 1 (IL-1R1). After observing protection against obesity-induced neuroinflammation and cognitive impairment in NLRP3-KO mice, we transplanted VAT from obese WT or NLRP3-KO donors into lean recipient mice. Transplantation of VAT from a WT donor (TRANSWT) increased hippocampal IL-1β and impaired cognition, but VAT transplants from comparably obese NLRP3-KO donors (TRANSKO) had no effect. Visceral adipose NLRP3 was required for deficits in long-term potentiation (LTP) in transplant recipients, and LTP impairment in TRANSWT mice was IL-1 dependent. Flow cytometric and gene expression analyses revealed that VAT transplantation recapitulated the effects of obesity on microglial activation and IL-1β gene expression, and visualization of hippocampal microglia revealed similar effects in vivo. Inducible ablation of IL-1R1 in CX3CR1-expressing cells eliminated cognitive impairment in mice with dietary obesity and in transplant recipients and restored immunoquiescence in hippocampal microglia. These results indicate that visceral adipose NLRP3 impairs memory via IL-1-mediated microglial activation and suggest that NLRP3/IL-1β signaling may underlie correlations between visceral adiposity and cognitive impairment in humans.


Genetic and proteomic analysis of factors affecting serum cholesterol reduction by Lactobacillus acidophilus A4.
This article identifies novel factors involved in cholesterol reduction by probiotic bacteria, which were identified using genetic and proteomic approaches. Approximately 600 Lactobacillus acidophilus A4 mutants were created by random mutagenesis. The cholesterol-reducing ability of each mutant was determined and verified using two different methods: the o-phthalaldehyde assay and gas chromatographic analysis (GC). Among screened mutants, strain BA9 showed a dramatically diminished ability to reduce cholesterol, as demonstrated by a 7.7% reduction rate, while the parent strain had a more than 50% reduction rate. The transposon insertion site was mapped using inverse PCR (I-PCR), and it was determined using bioinformatic methods that the deleted region contained the Streptococcus thermophilus catabolite control protein A gene (ccpA). In addition, we have shown using two-dimensional gel electrophoresis (2-DE) that several proteins, including a transcription regulator, FMN-binding protein, major facilitator superfamily permease, glycogen phosphorylase, the YknV protein, and fructose/tagatose bisphosphate aldolase, were strongly regulated by the ccpA gene. In addition, in vivo experiments investigating ccpA function were conducted with rats. Rats fed wild-type L. acidophilus A4 showed a greater than 20% reduction in total serum cholesterol, but rats fed BA9 mutant L. acidophilus showed only an approximately 10% reduction in cholesterol. These results provide important insights into the mechanism by which these lactic acid bacteria reduce cholesterol.


Healthier choices and increased participation in a middle school lunch program: effects of nutrition policy changes in San Francisco.
In order to address overall nutritional health, including increases in numbers of overweight children and adolescents, the San Francisco Unified School District implemented a progressive nutrition policy beginning in August 2003. We review this policy and focus on its impact on school and district revenues and students' participation in the federally subsidized school lunch program. We examined changes in menu items and the consequent effects of these changes on student eating patterns and school revenues at Aptos Middle School in San Francisco. This case study and additional district data show that provision of healthy menu options led to increased student participation in the federal school lunch program.


Gap solitons in grating superstructures.
We report results of the investigation of gap solitons (GSs) in the generic model of a periodically modulated Bragg grating (BG), which includes periodic modulation of the BG chirp or local refractive index, and periodic variation of the local reflectivity. We demonstrate that, while the previously studied reflectivity modulation strongly destabilizes all solitons, the periodic chirp modulation, which is a novel feature, stabilizes a new family of double-peak fundamental BGs in the side bandgap at negative frequencies (gap No. -1), and keeps solitons stable in the central bandgap (No. 0). The two soliton families demonstrate bistability, coexisting at equal values of energy. In addition, stable 4-peak bound states are formed by pairs of fundamental GSs in bandgap -1. Self-trapping and mobility of the solitons are studied too.


Effects of febuxostat on renal function in patients with chronic kidney disease: A systematic review and meta-analysis.
Hyperuricemia has been proven to be an independent risk factor for chronic kidney disease (CKD). However, the role of hyperuricemia in the progression of CKD remains unclear. Thus, we performed a systematic review and meta-analysis to evaluate the efficacy and safety of febuxostat, a first line urate-lowering agent, in CKD patients with hyperuricemia. We have systematically searched for randomized controlled trials assessing the efficacy and safety of febuxostat versus control in CKD patients with hyperuricemia through MEDLINE, PubMed, EMBASE, and Cochrane databases. All statistical analyses were conducted by using the statistical package Review Manager, version 5.3.5. Heterogeneity was assessed using the Cochrane Q and I tests and summary statistics were reported with 95% confidence interval. Two-tailed test was used for analysis and a P value of <.05 is considered statistically significant. Eleven eligible trials with 1317 participants were included in the meta-analysis. A significant reduction in serum uric acid was found in the febuxostat treated group. Also, a significant higher eGFR was found in the febuxostat treated group among CKD stage 3 and 4 patients. No significant difference of major complication or death was identified between treatment and control groups. The meta-analysis showed that other than its urate-lowering effect, febuxostat presented a reno-protective effect in CKD patients. More studies with larger sample sizes and higher quality are required to clarify the role of febuxostat use in the progression of CKD.


A bifunctional-biased mu-opioid agonist-neuropeptide FF receptor antagonist as analgesic with improved acute and chronic side effects.
Opioid analgesics, such as morphine, oxycodone, and fentanyl, are the cornerstones for treating moderate to severe pain. However, on chronic administration, their efficiency is limited by prominent side effects such as analgesic tolerance and dependence liability. Neuropeptide FF (NPFF) and its receptors (NPFF1R and NPFF2R) are recognized as an important pronociceptive system involved in opioid-induced hyperalgesia and analgesic tolerance. In this article, we report the design of multitarget peptidomimetic compounds that show high-affinity binding to the mu-opioid receptor (MOPr) and NPFFRs. In vitro characterization of these compounds led to identification of KGFF03 and KGFF09 as G-protein-biased MOPr agonists with full agonist or antagonist activity at NPFFRs, respectively. In agreement with their biased MOPr agonism, KGFF03/09 showed reduced respiratory depression in mice, as compared to the unbiased parent opioid agonist KGOP01. Chronic subcutaneous administration of KGOP01 and KGFF03 in mice rapidly induced hyperalgesia and analgesic tolerance, effects that were not observed on chronic treatment with KGFF09. This favorable profile was further confirmed in a model of persistent inflammatory pain. In addition, we showed that KGFF09 induced less physical dependence compared with KGOP01 and KGFF03. Altogether, our data establish that combining, within a single molecule, the G-protein-biased MOPr agonism and NPFFR antagonism have beneficial effects on both acute and chronic side effects of conventional opioid analgesics. This strategy can lead to the development of novel and potent antinociceptive drugs with limited side effects on acute and chronic administration.


Muscle acetylcholine receptor conversion into chloride conductance at positive potentials by a single mutation.
Charge selectivity forms the basis of cellular excitation or inhibition by Cys-loop ligand-gated ion channels (LGICs), and is essential for physiological receptor function. There are no reports of naturally occurring mutations in LGICs associated with the conversion of charge selectivity. Here, we report on a CHRNA1 mutation (α1Leu251Arg) in a patient with congenital myasthenic syndrome associated with transformation of the muscle acetylcholine receptor (AChR) into an inhibitory channel. Performing patch-clamp experiments, the AChR was found to be converted into chloride conductance at positive potentials, whereas whole-cell currents at negative potentials, although markedly reduced, were still carried by sodium. Umbrella sampling molecular dynamics simulations revealed constriction of the channel pore radius to 2.4 Å as a result of the mutation, which required partial desolvation of the ions in order to permeate the pore. Ion desolvation was associated with an energetic penalty that was compensated for by the favorable electrostatic interaction of the positively charged arginines with chloride. These findings reveal a mechanism for the transformation of the muscle AChR into an inhibitory channel in a clinical context.


Selenium supplementation for asthma.
Selenium deficiency may be important in chronic asthma. Observational studies have demonstrated that patients with chronic asthma may have lower levels of selenium than their control. Nevertheless, selenium supplementation has not been recommended with drug therapy for asthma. This review systematically examines RCTs that evaluated the role of selenium supplementation in chronic asthma. Recognition that chronic asthma can be associated with selenium deficiency has led to the investigation of the role of selenium supplementation in reducing the symptoms and impact of chronic asthma. The objective of this review was to assess the efficacy of selenium supplementation as an adjunct to medication for the treatment of chronic asthma. We searched the Cochrane Airways Group trials register, MEDLINE/PUB MED, and EMBASE. Searches were current as of August 2003. Randomised trials comparing patients with chronic asthma receiving selenium supplementation in conjunction with asthma medication, with patients taking asthma medication only. Two reviewers applied the study inclusion criteria One trial with a total of 24 patients suffering from chronic asthma was included. The study reported significant clinical improvement in the selenium-supplemented group, as compared with the placebo group, in terms of a 'clinical evaluation'. However, this improvement could not be validated by significant changes in separate objective parameters of lung function and airway hyper-responsiveness. There is some indication that selenium supplementation may be a useful adjunct to medication for patients with chronic asthma. This conclusion is limited because of insufficient studies and lack of improvement in the clinical parameters of lung function.


Contaminants and sea ducks in Alaska and the circumpolar region.
We review nesting sea duck population declines in Alaska during recent decades and explore the possibility that contaminants may be implicated. Aerial surveys of the surf scoter (Melanitta perspicillata), white-winged scoter (M. fusca), black scoter (M. nigra), oldsqaw (Clangula hyemalis), spectacled eider (Somateria fischeri), and Steller's eider (Polysticta stellei) show long-term breeding population declines, especially the latter three species. The spectacled eider was recently classified threatened under the Endangered Species Act. In addition, three other diving ducks, which commonly winter in coastal areas, have declined from unknown causes. Large die-offs of all three species of scoters during molt, a period of high energy demand, were documented in August 1990, 1991, and 1992 at coastal reefs in southeastern Alaska. There was no evidence of infectious diseases in those scoters. The die-offs may or may not be associated with the long-term declines. Many scoters had elevated renal concentrations of cadmium (high of 375 micrograms/g dry weight [dw]). Effects of cadmium in sea ducks are not well understood. Selenium concentrations in livers of nesting white-winged scoters were high; however, the eggs they laid contained less selenium than expected based on relationships for freshwater bird species. Histological evaluation found a high prevalence of hepatocellular vacuolation (49%), a degenerative change frequently associated with sublethal toxic insult. Cadmium and selenium mean liver concentrations were generally higher in those birds with more severe vacuolation; however, relationships were not statistically significant. We do not know if sea duck population declines are related to metals or other contaminants.


Mitochondrial DNA of the Mauritian macaques (Macaca fascicularis): an example of the founder effect.
Macaque monkeys (Macaca fascicularis) were introduced to the island of Mauritius approximately 400 years ago. This study compares the mitochondrial DNA of macaques on Mauritius with those from Indonesia and the Philippines. The goal is to measure the amount of evolutionary change that has occurred in this isolated population over 400 years, and to address questions regarding the origin of the Mauritian founders. Amplification of the control region of the mitochondrial genome via the polymerase chain reaction yielded an 1800 base pair DNA fragment which was surveyed for variation using restriction endonucleases. Fifty-two macaques were separated into 17 haplotypes by mapping the restriction sites. No haplotypes were shared among the three populations, and only two closely related haplotypes appeared in the Mauritian sample. Nucleotide variation in the mitochondrial DNA in the Mauritian sample was 10-fold less than the Indonesian and Filipino samples. In contrast, allozyme data estimates of genetic diversity on Mauritius are similar to populations from the ancestral range. The evidence of the more severe bottleneck as measured by mitochondrial data may be explained in part by almost exclusive male dispersal in this species, and may support models of founder events in which rapid population growth prevents substantial loss of nuclear variation. The mitochondrial evidence supports the morphologically and historically based hypothesis that the original founders came from Indonesia.


A Review of Plant-based Diets to Prevent and Treat Heart Failure.
Evidence supporting the role of nutrition in heart failure (HF) incidence and severity is growing. A comprehensive search of online databases was conducted using relevant keywords to identify human studies including diet and HF. Several plant-based diets have consistently been associated with decreased HF incidence and severity, notably the Dietary Approaches to Stop Hypertension (DASH) and Mediterranean diets. Several other plant-based dietary patterns, including low-fat diets and the rice diet, also show promise. Higher dietary quality, as assessed using different scores, seems to provide protective qualities. Fruit, vegetables, legumes and wholegrains appear to be beneficial, whereas red/processed meats, eggs and refined carbohydrates appear harmful. Some evidence suggests detrimental effects of dairy products and poultry, but more research is needed. There is observational and interventional evidence that a plant-based diet high in antioxidants, micronutrients, nitrate and fibre but low in saturated/trans fats may decrease the incidence and severity of HF. Potential mechanisms for this include decreased oxidative stress, homocysteine and inflammation levels, as well as higher antioxidant defence and nitric oxide bioavailability with gut microbiome modulation. Well-designed randomised, controlled nutrition intervention trials specific to HF are urgently required.


Investigating longitudinal associations between parent reported sleep in early childhood and teacher reported executive functioning in school-aged children with autism.
Up to 80% of children with autism spectrum disorder (ASD) experience sleep disturbance. Poor sleep impairs executive functioning (EF), a lifelong difficulty in ASD. Evidence suggests EF difficulties in ASD are exacerbated by poor sleep. We examine whether early childhood sleep disturbances are associated with worsening EF trajectories in school-aged children with ASD. A subsample (n = 217) from the Pathways in ASD longitudinal study was analyzed. The Children's Sleep Habits Questionnaire captured sleep duration, onset, and night awakenings before age 5 (mean = 3.5 years). Metacognition (MI) and Behavioral Regulation (BRI) indices, on the Teacher Behavior Rating Inventory of Executive Functioning, were used to measure cognitive and affective components of EF respectively at four time-points (7.8-11.8 years). We applied latent growth curve models to examine associations between sleep and EF, accounting for relevant covariates, including school-age sleep (mean = 6.7 years). Sleep traits had different age-related impacts on behavioral regulation, but not metacognition. Longer sleep onset at 3.5 years was associated with a worsening BRI difficulties slope (b = 2.07, p < 0.04), but conversely associated with lower BRI difficulties at 7.7 years (b = -4.14, p = 0.04). A longer sleep onset at 6.7 years was related to higher BRI difficulties at 7.7 years (b = 7.78, p < 0.01). Longer sleep duration at 6.7 years was associated with higher BRI difficulties at age 7.7 (b = 3.15, p = 0.01), but subscale analyses revealed shorter sleep duration at age 6.7 was linked to a worsening inhibition slope (b = -0.60, p = 0.01). Sleep onset is a robust early correlate of behavior regulation in children with ASD, whereas sleep duration is a later childhood correlate.


Study of the ground multiplet of Kramers rare earth ions in solid matrices by multifrequency electron paramagnetic resonance spectroscopy: Nd3+ in PbWO4 single-crystals.
A multifrequency electron paramagnetic resonance (EPR) investigation of Nd(3+) impurities in PbWO(4) single-crystals at the conventional microwave frequency (MF) 9.43 GHz, and at the 95, 190, and 285 GHz high frequencies was carried out. The resulting spectra are well described at all frequencies by an axial spin-Hamiltonian corresponding to an effective electron spin of one-half and to a tetragonal symmetry. For the magnetic field along the tetragonal axis, the g(parallel)-factor and the hyperfine constant A(parallel) of the lowest doublet of the ground multiplet decreases with frequency increase. For the magnetic field perpendicular to the tetragonal axis, the g(perpendicular)-factor exhibits a small azimuthal angular dependence that increases with increasing the frequency due to the S(4) site symmetry. The azimuthal angular dependence allows to clearly distinguish between different local axial symmetries. These properties are interpreted as high field/frequency (HF) effects associated with the mixing by the large Zeeman interaction of some of the upper-lying doublets of the ground multiplet into the lowest-lying doublet states. We show that from the combined analysis of the multifrequency MF- and HF-EPR spectra and of the optical data, an accurate description of the ground multiplet of the Kramers rare earth ions in solid matrices can be derived.


Epidemiological and molecular investigation of a rubella outbreak, Romania, 2011 to 2012.
We describe a rubella outbreak that occurred in Romania between September 2011 and December 2012. During this period 24,627 rubella cases, 41.1% (n=10,134) of which female, were notified based on clinical criteria, and a total of 6,182 individuals were found serologically positive for IgM-specific rubella antibody. The median age of notified cases was 18 years (range: <1-65) and the most affected age group 15 to 19 years (n=16,245 cases). Of all notified cases, 24,067 cases (97.7%) reported no history of vaccination. Phylogenetic analysis of 19 sequences (739 nucleotides each), from 10 districts of the country revealed that the outbreak was caused by two distinct rubella virus strains of genotype 2B, which co-circulated with both temporal and geographical overlap. In addition to the 6,182 IgM-positive rubella cases, 28 cases of congenital rubella syndrome (CRS) were identified, including 11 neonatal deaths and one stillbirth. The outbreak underscores the need to encourage higher vaccination uptake in the population, particularly in women of reproductive age, and to strengthen epidemiological and laboratory investigations of suspected rubella cases. Genetic characterisation of wild-type rubella virus is an essential component to enhance surveillance and here we report rubella virus sequences from Romania.


Far-red fluorescent tags for protein imaging in living tissues.
A vast colour palette of monomeric fluorescent proteins has been developed to investigate protein localization, motility and interactions. However, low brightness has remained a problem in far-red variants, which hampers multicolour labelling and whole-body imaging techniques. In the present paper, we report mKate2, a monomeric far-red fluorescent protein that is almost 3-fold brighter than the previously reported mKate and is 10-fold brighter than mPlum. The high-brightness, far-red emission spectrum, excellent pH resistance and photostability, coupled with low toxicity demonstrated in transgenic Xenopus laevis embryos, make mKate2 a superior fluorescent tag for imaging in living tissues. We also report tdKatushka2, a tandem far-red tag that performs well in fusions, provides 4-fold brighter near-IR fluorescence compared with mRaspberry or mCherry, and is 20-fold brighter than mPlum. Together, monomeric mKate2 and pseudo-monomeric tdKatushka2 represent the next generation of extra-bright far-red fluorescent probes offering novel possibilities for fluorescent imaging of proteins in living cells and animals.


Use of oxycodone in polytrauma patients: the "Gemelli" experience.
This is the first study investigating the effect of oxycodone in polytrauma patients. The management of pain in polytrauma patients has become a very relevant issue. Nonsteroidal anti-inflammatory drugs (NSAIDs) represent the most used drugs in polytrauma patients, even though their use is associated with an increased hemorrhagic risk. Previous studies have demonstrated the efficacy of oxycodone for the treatment of acute pain. The aim of this study was to assess the efficacy of oxycodone administration in polytrauma patients, with minor injuries. 15 polytrauma patients (10 males, mean age 40 +/- 13 years; 5 females, mean age 49 +/- 26 years) were admitted to the Emergency Department of the Catholic University, A. Gemelli Hospital in Rome, Italy. All patients underwent physical examination, FAST ultrasound, total body CT scanning and blood tests. Three patients had multiple costal fractures, three had pelvic fracture, two had tibial fracture, five had vertebral fractures, one patient had clavicle fracture and ulnar fracture, one patient a severe trauma of the left leg, which required amputation. Five patients also reported minor head trauma, with a Glasgow Coma Score (GCS) 15. All patients reported abdominal trauma, while none of them had severe thoracic or kidney damage. Patients with head trauma also underwent a second CT head scanning 12 hours after admission, which excluded the occurrence of cerebral damage. All patients were then treated with oral administration of oxycodone 10 mg two times per day (bid) for 3 days. Pain intensity, before and after the administration of oxycodone, was evaluated using a scale ranging from 0 to 10. The mean pain score at admission was 8 +/- 0.7. All patients reported significant pain improvement after the administration of oxycodone (8 +/- 0.7 vs 1.8 +/- 0.9; p < 0.0001). A dosage increase of oxycodone from 20 to 40 mg bid was required in only one patient with a clavicle fracture. The main side effects were light-headache (5 patients), constipation (4 patients) and nausea (3 patients). These data indicate that oxycodone is a safe and effective drug for pain relief in polytrauma patients without severe thoracic, kidney or brain damage.


Global analysis of the yeast lipidome by quantitative shotgun mass spectrometry.
Although the transcriptome, proteome, and interactome of several eukaryotic model organisms have been described in detail, lipidomes remain relatively uncharacterized. Using Saccharomyces cerevisiae as an example, we demonstrate that automated shotgun lipidomics analysis enabled lipidome-wide absolute quantification of individual molecular lipid species by streamlined processing of a single sample of only 2 million yeast cells. By comparative lipidomics, we achieved the absolute quantification of 250 molecular lipid species covering 21 major lipid classes. This analysis provided approximately 95% coverage of the yeast lipidome achieved with 125-fold improvement in sensitivity compared with previous approaches. Comparative lipidomics demonstrated that growth temperature and defects in lipid biosynthesis induce ripple effects throughout the molecular composition of the yeast lipidome. This work serves as a resource for molecular characterization of eukaryotic lipidomes, and establishes shotgun lipidomics as a powerful platform for complementing biochemical studies and other systems-level approaches.


Sympathetic neural mechanisms in obstructive sleep apnea.
Blood pressure, heart rate, sympathetic nerve activity, and polysomnography were recorded during wakefulness and sleep in 10 patients with obstructive sleep apnea. Measurements were also obtained after treatment with continuous positive airway pressure (CPAP) in four patients. Awake sympathetic activity was also measured in 10 age- and sex-matched control subjects and in 5 obese subjects without a history of sleep apnea. Patients with sleep apnea had high levels of nerve activity even when awake (P < 0.001). Blood pressure and sympathetic nerve activity did not fall during any stage of sleep. Mean blood pressure was 92 +/- 4.5 mmHg when awake and reached peak levels of 116 +/- 5 and 127 +/- 7 mmHg during stage II sleep (n = 10) and rapid eye movement (REM) sleep (n = 5), respectively (P < 0.001). Sympathetic activity increased during sleep (P = 0.01) especially during stage II (133 +/- 9% above wakefulness; P = 0.006) and REM (141 +/- 13%; P = 0.007). Peak sympathetic activity (measured over the last 10 s of each apneic event) increased to 299 +/- 96% during stage II sleep and to 246 +/- 36% during REM sleep (both P < 0.001). CPAP decreased sympathetic activity and blood pressure during sleep (P < 0.03). We conclude that patients with obstructive sleep apnea have high sympathetic activity when awake, with further increases in blood pressure and sympathetic activity during sleep. These increases are attenuated by treatment with CPAP.


Deep sequencing analysis of small noncoding RNA and mRNA targets of the global post-transcriptional regulator, Hfq.
Recent advances in high-throughput pyrosequencing (HTPS) technology now allow a thorough analysis of RNA bound to cellular proteins, and, therefore, of post-transcriptional regulons. We used HTPS to discover the Salmonella RNAs that are targeted by the common bacterial Sm-like protein, Hfq. Initial transcriptomic analysis revealed that Hfq controls the expression of almost a fifth of all Salmonella genes, including several horizontally acquired pathogenicity islands (SPI-1, -2, -4, -5), two sigma factor regulons, and the flagellar gene cascade. Subsequent HTPS analysis of 350,000 cDNAs, derived from RNA co-immunoprecipitation (coIP) with epitope-tagged Hfq or control coIP, identified 727 mRNAs that are Hfq-bound in vivo. The cDNA analysis discovered new, small noncoding RNAs (sRNAs) and more than doubled the number of sRNAs known to be expressed in Salmonella to 64; about half of these are associated with Hfq. Our analysis explained aspects of the pleiotropic effects of Hfq loss-of-function. Specifically, we found that the mRNAs of hilD (master regulator of the SPI-1 invasion genes) and flhDC (flagellar master regulator) were bound by Hfq. We predicted that defective SPI-1 secretion and flagellar phenotypes of the hfq mutant would be rescued by overexpression of HilD and FlhDC, and we proved this to be correct. The combination of epitope-tagging and HTPS of immunoprecipitated RNA detected the expression of many intergenic chromosomal regions of Salmonella. Our approach overcomes the limited availability of high-density microarrays that have impeded expression-based sRNA discovery in microorganisms. We present a generic strategy that is ideal for the systems-level analysis of the post-transcriptional regulons of RNA-binding proteins and for sRNA discovery in a wide range of bacteria.


Amplified inhibition of stellate cell activation pathways by PPAR-γ, RAR and RXR agonists.
Peroxisome proliferator activator receptors (PPAR) ligands such as 15-Δ12,13-prostaglandin L(2) [PJ] and all trans retinoic acid (ATRA) have been shown to inhibit the development of liver fibrosis. The role of ligands of retinoic X receptor (RXR) and its ligand, 9-cis, is less clear. The purpose of this study was to investigate the effects of combined treatment of the three ligends, PJ, ATRA and 9-cis, on key events during liver fibrosis in rat primary hepatic stellate cells (HSCs). We found that the anti-proliferative effect of the combined treatment of PJ, ATRA and 9-cis on HSCs was additive. Further experiments revealed that this inhibition was due to cell cycle arrest at the G0/G1 phase as demonstrated by FACS analysis. In addition, the combined treatment reduced cyclin D1 expression and increased p21 and p27 protein levels. Furthermore, we found that the three ligands down regulated the phosphorylation of mTOR and p70(S6K). The activation of HSCs was also inhibited by the three ligands as shown by inhibition of vitamin A lipid droplets depletion from HSCs. Studies using real time PCR and western blot analysis showed marked inhibition of collagen Iα1 and αSMA by the combination of the three ligands. These findings suggest that the combined use of PJ, ATRA and 9-cis causes inhibition of cell proliferation by cell cycle arrest and down-regulation of fibrotic markers to a greater extent compared to each of the ligands alone.


Molecular systematic and historical biogeography of the armored Neotropical catfishes Hypoptopomatinae and Neoplecostominae (Siluriformes: Loricariidae).
The Neotropics possess the greatest freshwater fish diversity of the world, rendering the study of their evolutionary history extremely challenging. Loricariidae catfishes are one of the most diverse components of the Neotropical ichthyofauna and despite a long history of classification, major issues still need elucidation. Based on a nuclear gene, we present a robust phylogeny of two former loricariid subfamilies: Hypoptopomatinae and Neoplecostominae. Our results show that Neoplecostominae is nested within Hypoptopomatinae, and is the sister group to the former Otothyrini tribe. According to our results, supplemented by morphological observations, we erect two new subfamilies, the Otothyrinae and a new Hypoptopomatinae, and modify the Neoplecostominae by including the genus Pseudotocinclus. The uncovered evolutionary relationships allow a detailed analysis of their historical biogeography. We tested two Dispersal-Extinction-Cladogenesis models for inferring the distribution range evolution of the new subfamilies, and show that the model having no constrains performs better than a model constraining long-range dispersal. The Maximum Likelihood reconstructions of ancestral ranges showed a marked division between the Amazonian origin of the Hypoptopomatinae and the eastern coastal Brazil+Upper Paraná origin of the Neoplecostominae and Otothyrinae. Markedly few instances of dispersal across the border separating the Amazon basin and the Paraná-Paraguay+eastern coastal Brazil+Uruguay were reconstructed. This result is in clear contrast with the historical biogeography of many Neotropical fishes, including other Loricariidae. Part of the dispersal limitation may be explained by divergent ecological specialization: lowland rivers versus mountain streams habitats. Moreover, because most species of the new subfamilies are small, we hypothesize that body size-related effects might limit their dispersal, like predation and energetic cost to migration. Finally, morphological and anatomical features are presented that limit or, to the contrary, enhance dispersal capability in these small and fascinating catfishes.


Global, Regional, and National Cancer Incidence, Mortality, Years of Life Lost, Years Lived With Disability, and Disability-Adjusted Life-Years for 29 Cancer Groups, 1990 to 2017: A Systematic Analysis for the Global Burden of Disease Study.
Cancer and other noncommunicable diseases (NCDs) are now widely recognized as a threat to global development. The latest United Nations high-level meeting on NCDs reaffirmed this observation and also highlighted the slow progress in meeting the 2011 Political Declaration on the Prevention and Control of Noncommunicable Diseases and the third Sustainable Development Goal. Lack of situational analyses, priority setting, and budgeting have been identified as major obstacles in achieving these goals. All of these have in common that they require information on the local cancer epidemiology. The Global Burden of Disease (GBD) study is uniquely poised to provide these crucial data. To describe cancer burden for 29 cancer groups in 195 countries from 1990 through 2017 to provide data needed for cancer control planning. We used the GBD study estimation methods to describe cancer incidence, mortality, years lived with disability, years of life lost, and disability-adjusted life-years (DALYs). Results are presented at the national level as well as by Socio-demographic Index (SDI), a composite indicator of income, educational attainment, and total fertility rate. We also analyzed the influence of the epidemiological vs the demographic transition on cancer incidence. In 2017, there were 24.5 million incident cancer cases worldwide (16.8 million without nonmelanoma skin cancer [NMSC]) and 9.6 million cancer deaths. The majority of cancer DALYs came from years of life lost (97%), and only 3% came from years lived with disability. The odds of developing cancer were the lowest in the low SDI quintile (1 in 7) and the highest in the high SDI quintile (1 in 2) for both sexes. In 2017, the most common incident cancers in men were NMSC (4.3 million incident cases); tracheal, bronchus, and lung (TBL) cancer (1.5 million incident cases); and prostate cancer (1.3 million incident cases). The most common causes of cancer deaths and DALYs for men were TBL cancer (1.3 million deaths and 28.4 million DALYs), liver cancer (572 000 deaths and 15.2 million DALYs), and stomach cancer (542 000 deaths and 12.2 million DALYs). For women in 2017, the most common incident cancers were NMSC (3.3 million incident cases), breast cancer (1.9 million incident cases), and colorectal cancer (819 000 incident cases). The leading causes of cancer deaths and DALYs for women were breast cancer (601 000 deaths and 17.4 million DALYs), TBL cancer (596 000 deaths and 12.6 million DALYs), and colorectal cancer (414 000 deaths and 8.3 million DALYs). The national epidemiological profiles of cancer burden in the GBD study show large heterogeneities, which are a reflection of different exposures to risk factors, economic settings, lifestyles, and access to care and screening. The GBD study can be used by policy makers and other stakeholders to develop and improve national and local cancer control in order to achieve the global targets and improve equity in cancer care.


Integration of HIV services with primary care in Yangon, Myanmar: a retrospective cohort analysis.
Integration of HIV care with general healthcare may improve patient engagement. We assessed patient outcomes in four clinics offering HIV care integrated into primary care clinics in Yangon, Myanmar. We carried out a retrospective cohort analysis of 4551 patients who started antiretroviral therapy between 2009 and 2017. Mortality and disengagement from care were assessed using Cox regression. People living with HIV presented late with low CD4 counts [median (25th , 75th percentile) = 178 (65, 308) from 4216 patients] and advanced HIV (69% with stage 3 or 4). Survival was 0.95 at 1 year and 0.90 at 5 years. Males were at a higher risk of mortality than females [unadjusted hazard ratio (uHR) = 1.6 (95% CI: 1.3-2.0). Patients linked to HIV care via antenatal care or partner/parent notification were at reduced risk of mortality [uHR = 0.4 (95% CI: 0.1-1.0) and uHR = 0.5 (95% CI: 0.3-0.7)] relative to patients who presented for HIV testing. The cumulative incidence of disengagement was 0.06 at 1 year and 0.15 at 5 years. Young adults had a higher risk of disengagement than did children and older patients. Women linked to HIV care via antenatal care services were at increased risk of disengagement relative to patients who came for HIV testing (uHR = 2.4; 95% CI: 1.7-3.4). Mortality and disengagement remained steady over calendar time as the programme scaled up. HIV care within a primary care model is effective to attain early linkage to care, with high survival. However, close attention should be given to disengagement from care, in particular for pregnant women.


Propagation of mouse and human T cells with defined antigen specificity and function.
Difficulties maintaining fully functional CD4+ T cells in culture have historically limited the study of their role in tumour rejection as well as other clinical applications. As the therapeutic value of current antitumour CD8+ T cell adoptive therapy becomes better defined, a strong impetus exists to determine optimal conditions for culturing antitumour CD4+ T cells. Our goal is to promote broadly polyclonal, antigen-specific CD4+ T cell responses of either Th1 or Th2 character for use in antitumour therapy or allograft facilitation, respectively. Similar obstacles exist in murine and human cultures: (1) during even brief periods of culture CD4+ T cells develop high 'background' reactivity to class II-positive antigen-presenting cells; (2) maintenance of antigen specificity as evidenced by cytokine secretion and short-term proliferation assays is insufficient to ensure bulk numerical expansion; (3) Th1-type CD4+ T cells often lose their potential for antigen-specific secretion of interleukin 2 on re-stimulation (though remain inducible by 12-O-tetradecanoylphorbol 13-acetate/ionomycin); (4) during prolonged culture selection pressure favours CD4+ subpopulations that recognize artifactual antigens such as culture medium proteins; (5) even with optimal culture conditions, cultured CD4+ T cells may function differently in vivo to uncultured CD4+ T cells. We have devised various strategies to surmount these obstacles by use of selected cytokines, antigen-presenting cells and timely culture manoeuvres.


Correlation matching method for high-precision position detection of optical vortex using Shack-Hartmann wavefront sensor.
We propose a new method for realizing high-spatial-resolution detection of singularity points in optical vortex beams. The method uses a Shack-Hartmann wavefront sensor (SHWS) to record a Hartmanngram. A map of evaluation values related to phase slope is then calculated from the Hartmanngram. The position of an optical vortex is determined by comparing the map with reference maps that are calculated from numerically created spiral phases having various positions. Optical experiments were carried out to verify the method. We displayed various spiral phase distribution patterns on a phase-only spatial light modulator and measured the resulting singularity point using the proposed method. The results showed good linearity in detecting the position of singularity points. The RMS error of the measured position of the singularity point was approximately 0.056, in units normalized to the lens size of the lenslet array used in the SHWS.


Temperature as a Causative Factor in Diabetic Foot Ulcers: A Call to Revisit Ulceration Pathomechanics.
Diabetic foot ulcers (DFUs) are a major burden to patients and to the health-care systems of many countries. To prevent or treat ulcers more effectively, predictive biomarkers are needed. We examined temperature as a biomarker and as a causative factor in ulcer development. Thirty-seven individuals with diabetes were enrolled in this observational case-control study: nine with diabetic neuropathy and ulcer history (DFU), 14 with diabetic neuropathy (DN), and 14 nonneuropathic control participants (DC). Resting barefoot plantar temperatures were recorded using an infrared thermal camera. Mean temperatures were determined in four anatomical regions-hallux and medial, central, and lateral forefoot-and separate linear models with specified contrasts among the DFU, DN, and DC groups were set to reveal mean differences for each foot region while controlling for group characteristics. The mean temperature reading in each foot region was higher than 30.0°C in the DFU and DN groups and lower than 30.0°C in the DC group. Mean differences were greatest between the DFU and DC groups, ranging from 3.2°C in the medial forefoot to 4.9°C in the hallux. Increased plantar temperatures in individuals with a history of ulcers may include acute temperature increases from plantar stresses, chronic inflammation from prolonged stresses, and impairment in temperature regulation from autonomic neuropathy. Diabetic foot temperatures, particularly in patients with previous ulcers, may easily reach hazard thresholds indicated by previous pressure ulcer studies. The results necessitate further exploration of temperature in the diabetic foot and how it may contribute to ulceration.


Targeting Immunity in End-Stage Renal Disease.
Despite the stable incidence of end-stage renal disease (ESRD), it continues to be associated with an unacceptably high cardiovascular risk. ESRD is characterized by enhanced oxidative stress and severe inflammation, which boost cardiovascular risk, thus increasing cardiovascular-associated mortality rate. While substantial effort has been made in the technological innovation of dialytic techniques, few significant advances have been made to reduce inflammation in patients with ESRD. Indeed, this contrasts with the extensive scientific breakthroughs made in the basic field of science in targeting inflammation. There is thus a pressing need for clinical trials to test the effect of reducing inflammation in patients with ESRD. Here, we will revisit the negative effect of ESRD on inflammation and explore the impact of enhanced inflammation on cardiovascular outcomes and survival in patients with ESRD. Finally, we will discuss the need for clinical trials that target inflammation in ESRD, as well as weigh potential disadvantages and offer novel innovative approaches. Key Message: We will try to understand why the issue of inflammation has not been successfully addressed thus far in patients with ESRD, while at the same time weighing the potential disadvantages and offering novel innovative approaches for targeting inflammation in patients with ESRD.


Parallel Evolution of Group B Streptococcus Hypervirulent Clonal Complex 17 Unveils New Pathoadaptive Mutations.
Group B Streptococcus (GBS) is a commensal of the gastrointestinal and genitourinary tracts, while a prevailing cause of neonatal disease worldwide. Of the various clonal complexes (CCs), CC17 is overrepresented in GBS-infected newborns for reasons that are still largely unknown. Here, we report a comprehensive genomic analysis of 626 CC17 isolates collected worldwide, identifying the genetic traits behind their successful adaptation to humans and the underlying differences between carriage and clinical strains. Comparative analysis with 923 GBS genomes belonging to CC1, CC19, and CC23 revealed that the evolution of CC17 is distinct from that of other human-adapted lineages and recurrently targets functions related to nucleotide and amino acid metabolism, cell adhesion, regulation, and immune evasion. We show that the most distinctive features of disease-specific CC17 isolates were frequent mutations in the virulence-associated CovS and Stk1 kinases, underscoring the crucial role of the entire CovRS regulatory pathway in modulating the pathogenicity of GBS. Importantly, parallel and convergent evolution of major components of the bacterial cell envelope, such as the capsule biosynthesis operon, the pilus, and Rib, reflects adaptation to host immune pressures and should be taken into account in the ongoing development of a GBS vaccine. The presence of recurrent targets of evolution not previously implicated in virulence also opens the way for uncovering new functions involved in host colonization and GBS pathogenesis. IMPORTANCE The incidence of group B Streptococcus (GBS) neonatal disease continues to be a significant cause of concern worldwide. Strains belonging to clonal complex 17 (CC17) are the most frequently responsible for GBS infections in neonates, especially among late-onset disease cases. Therefore, we undertook the largest genomic study of GBS CC17 strains to date to decipher the genetic bases of their remarkable colonization and infection ability. We show that crucial functions involved in different steps of the colonization or infection process of GBS are distinctly mutated during the adaptation of CC17 to the human host. In particular, our results implicate the CovRS two-component regulator of virulence in the differentiation between carriage- and disease-associated isolates. Not only does this work raise important implications for the ongoing development of a vaccine against GBS but might also drive the discovery of key functions for GBS adaptation and pathogenesis that have been overlooked until now. Author Video: An author video summary of this article is available.


Particle size measurement from infrared laser ablation of tissue.
The concentration and size distribution were measured for particles ablated from tissue sections using an infrared optical parametric oscillator laser system. A scanning mobility particle sizer and light scattering particle sizer were used in parallel to realize a particle sizing range from 10 nm to 20 μm. Tissue sections from rat brain and lung ranging in thickness between 10 and 50 μm were mounted on microscope slides and irradiated with nanosecond laser pulses at 3 μm wavelength and fluences between 7 and 21 kJ m(-2) in reflection geometry. The particle size distributions were characterized by a bimodal distribution with a large number of particles 100 nm in diameter and below and a large mass contribution from particles greater than 1 μm in diameter. The large particle contribution dominated the ablated particle mass at high laser fluence. The tissue type, thickness, and water content did not have a significant effect on the particle size distributions. The implications of these results for laser ablation sampling and mass spectrometry imaging under ambient conditions are discussed.


De Novo Assembly of the Whole Transcriptome of the Wild Embryo, Preleptocephalus, Leptocephalus, and Glass Eel of Anguilla japonica and Deciphering the Digestive and Absorptive Capacities during Early Development.
Natural stocks of Japanese eel (Anguilla japonica) have decreased drastically because of overfishing, habitat destruction, and changes in the ocean environment over the past few decades. However, to date, artificial mass production of glass eels is far from reality because of the lack of appropriate feed for the eel larvae. In this study, wild glass eel, leptocephali, preleptocephali, and embryos were collected to conduct RNA-seq. Approximately 279 million reads were generated and assembled into 224,043 transcripts. The transcript levels of genes coding for digestive enzymes and nutrient transporters were investigated to estimate the capacities for nutrient digestion and absorption during early development. The results showed that the transcript levels of protein digestion enzymes were higher than those of carbohydrate and lipid digestion enzymes in the preleptocephali and leptocephali, and the transcript levels of amino acid transporters were also higher than those of glucose and fructose transporters and the cholesterol transporter. In addition, the transcript levels of glucose and fructose transporters were significantly raising in the leptocephali. Moreover, the transcript levels of protein, carbohydrate, and lipid digestion enzymes were balanced in the glass eel, but the transcript levels of amino acid transporters were higher than those of glucose and cholesterol transporters. These findings implied that preleptocephali and leptocephali prefer high-protein food, and the nutritional requirements of monosaccharides and lipids for the eel larvae vary with growth. An online database (http://molas.iis.sinica.edu.tw/jpeel/) that will provide the sequences and the annotated results of assembled transcripts was established for the eel research community.


A cohesin/HUSH- and LINC-dependent pathway controls ribosomal DNA double-strand break repair.
The ribosomal DNA (rDNA) represents a particularly unstable locus undergoing frequent breakage. DNA double-strand breaks (DSBs) within rDNA induce both rDNA transcriptional repression and nucleolar segregation, but the link between the two events remains unclear. Here we found that DSBs induced on rDNA trigger transcriptional repression in a cohesin- and HUSH (human silencing hub) complex-dependent manner throughout the cell cycle. In S/G2 cells, transcriptional repression is further followed by extended resection within the interior of the nucleolus, DSB mobilization at the nucleolar periphery within nucleolar caps, and repair by homologous recombination. We showed that nuclear envelope invaginations frequently connect the nucleolus and that rDNA DSB mobilization, but not transcriptional repression, involves the nuclear envelope-associated LINC complex and the actin pathway. Altogether, our data indicate that rDNA break localization at the nucleolar periphery is not a direct consequence of transcriptional repression but rather is an active process that shares features with the mobilization of persistent DSB in active genes and heterochromatin.


18F-FDG PET/CT: Therapy Response Assessment Interpretation (Hopkins Criteria) and Survival Outcomes in Lung Cancer Patients.
The purpose of this study was to evaluate the value of an (18)F-FDG PET/CT-based interpretation system (Hopkins criteria) to assess the therapy response and survival in lung cancer. This is an Institutional Review Board-approved, retrospective study. A total of 201 patients with biopsy-proven lung cancer, who underwent therapy assessment (18)F-FDG PET/CT within 6 mo (mean, 7.5 wk) of completion of treatment, were included. Patients were primarily treated with surgical resection, chemotherapy, radiation therapy, or a combination of these treatments. PET/CT studies were interpreted by 2 nuclear medicine physicians, and discrepancies were resolved by a third interpreter. The studies were scored using a qualitative 5-point scale for the primary tumor, mediastinum, distant metastatic site, if present, and overall assessment. Scores 1, 2, and 3 were considered negative and scores 4 and 5 were considered positive for residual disease. Patients were followed for a median of 12 mo (up to 128 mo). Kaplan-Meier plots with a Mantel-Cox log-rank test were performed considering death as the endpoint. Overall, the PET/CT studies were positive in 144 (71.6%) and negative in 57 (28.4%) patients. There was substantial agreement between 2 interpreters (R1, R2), with a κ of 0.78 (P < 0.001). The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the Hopkins scoring system were 89%, 80%, 92.8%, 71.4%, and 86.7%, respectively. Overall, PET/CT resulted in starting a new treatment plan in 70.8% of patients with positive residual disease on therapy assessment PET/CT. There was a significant difference in overall survival (OS) between patients who were categorized as positive in comparison to those who were categorized as negative (hazard ratio [HR] = 2.12; 95% confidence interval = 1.44-3.12), which remained significant after adjustment for disease stage, prior clinical suspicion, and primary treatment. Subgroup analysis according to the tumor histology showed that positive Hopkins scoring could significantly predict the OS in both small cell lung cancer (HR = 2.88; log-rank, P = 0.02) and non-small cell lung cancer (HR = 2.01; log-rank, P = 0.001). Similarly, there was a significant difference in OS between patients with positive and negative Hopkins score both in those who had surgical resection as part of the primary treatment (HR = 6.09; log-rank, P < 0.001) and in those who were treated with chemotherapy with or without radiation (HR = 1.60; log-rank, P = 0.02). The 5-point qualitative therapy response interpretation for lung cancer has substantial interinterpreter agreement and high accuracy and could significantly predict survival in lung cancer, irrespective of tumor histology and treatment modality.


Pulmonary-renal syndromes: an update for respiratory physicians.
Pulmonary-renal syndromes are a group of disorders characterised by necrotising glomerulonephritis and pulmonary haemorrhage. Small vessel systemic vasculitis is the most common cause of pulmonary-renal syndromes presenting to respiratory physicians. Rarer causes include systemic lupus erythematosus and connective tissue diseases though severe pneumonia or cardiac failure may mimic their presentation. Some forms of small vessel vasculitides have a predilection for the pulmonary and renal vascular beds and if left untreated can result in fulminant organ failure. Whilst the aetiology of these syndromes remains unclear, much is known about the disease mechanisms including the pathogenic role of autoantibodies, immune-complex mediated inflammation and microangiopathic in-situ thrombosis. Despite established treatments achieving successful remission induction, patient tolerability and side effect profiles have limited their use which has led to searches for more targeted treatments. Consequently newer biological therapies have gained wider acceptance despite little being known about their long term safety and efficacy. The European Vasculitis Study Group (EUVAS) have recently formulated guidelines to provide consensus on diagnosis and management in this area and work to define survival rates in these conditions with longer term follow-up studies is ongoing. This review summarises the current aetiopathogenesis thought to underlie these complex diseases, the diagnostic definitions and classification criteria currently in use and the evidence base for modern therapies. Though unusual for respiratory specialists to coordinate overall management of these patients, an update on their current management is regarded as important to their practice given the recently changing trends in treatments.


Detection of freely propagating terahertz radiation by use of optical second-harmonic generation.
We report the application of electric-field-induced optical second-harmonic generation as a new technique for measuring the field of freely propagating terahertz radiation. Using silicon as the nonlinear medium, we demonstrate subpicosecond time resolution and a sampling signal that varies linearly with the terahertz electric field. This approach, which is attractive for centrosymmetric media, permits a significantly broadened class of materials to be exploited for free-space sampling measurements.


Genome sequence of Jatropha curcas L., a non-edible biodiesel plant, provides a resource to improve seed-related traits.
Jatropha curcas (physic nut), a non-edible oilseed crop, represents one of the most promising alternative energy sources due to its high seed oil content, rapid growth and adaptability to various environments. We report ~339 Mbp draft whole genome sequence of J. curcas var. Chai Nat using both the PacBio and Illumina sequencing platforms. We identified and categorized differentially expressed genes related to biosynthesis of lipid and toxic compound among four stages of seed development. Triacylglycerol (TAG), the major component of seed storage oil, is mainly synthesized by phospholipid:diacylglycerol acyltransferase in Jatropha, and continuous high expression of homologs of oleosin over seed development contributes to accumulation of high level of oil in kernels by preventing the breakdown of TAG. A physical cluster of genes for diterpenoid biosynthetic enzymes, including casbene synthases highly responsible for a toxic compound, phorbol ester, in seed cake, was syntenically highly conserved between Jatropha and castor bean. Transcriptomic analysis of female and male flowers revealed the up-regulation of a dozen family of TFs in female flower. Additionally, we constructed a robust species tree enabling estimation of divergence times among nine Jatropha species and five commercial crops in Malpighiales order. Our results will help researchers and breeders increase energy efficiency of this important oil seed crop by improving yield and oil content, and eliminating toxic compound in seed cake for animal feed.


A Case Report of Synchronous Prostate Cancer and Rectal Gastrointestinal Stromal Tumor and its Management.
A 72-year-old Japanese man presented to the hospital with complaints of gross hematuria. Contrast enhanced com-puted tomography revealed a broad-based, approximately 3-cm bladder tumor near the right ureteral orifice and a 5-cm mass located between the prostate and rectum. The patient underwent transurethral resection of the bladder tumor and a transrectal biopsy of the lesion. Histological examination of the specimens suggested that the patient had a muscle invasive adenocarcinoma from transurethral resection and a rectal gastrointestinal stromal tumor in TRUS biopsy. Patient underwent total pelvic exenteration and ileal and colonic stomas to divert urine and faeces. Final pathology report of the resected specimen revealed a rectal gastrointestinal stromal tumor and prostatic ad-enocarcinoma, which had invaded the urinary bladder and seminal vesicles. Synchronous gastrointestinal stromal tumor in rectum and prostate cancer treated with total pelvic exenteration has not been reported before in literature.


Demographic determinants of survival in osteosarcoma.
Osteosarcoma treatment has experienced a renaissance in the last 3 decades with the institution of multimodality treatment involving multiagent chemotherapy and surgery. Yet globally, treatment success has stagnated at about 70% survival at 5 years in most single institution series. We performed survival analyses on 2 national databases in 2 countries and compared these with corresponding institution specific survival. All patients with the diagnostic code of non-metastatic intramedullary osteosarcoma in the long bones of the upper and lower limbs less than 30 years of age were selected from the Surveillance Epidemiology and End Result (SEER) database to ensure uniformity with respect to disease and treatment. We studied the factors: ethnicity, gender, age, grade, histology, size, site, surgery, compartmentalisation, number of primaries and venue of treatment for their contribution to survival. In addition, the data were stratified into 3 decades (seventies, eighties and nineties) to account for variations due to the evolution of treatment paradigms and imaging modalities. Institution-specific survival was predictably better than national survival in the 4 databases. One thousand patients were selected from the SEER database. Oriental descent, state-specific treatment, female gender, treatment in the nineties, low-grade disease, intra-compartmental disease, small size, wide resections as opposed to forequarter or hindquarter amputations, and single primaries were good prognostic factors on univariate analysis as well as multivariate analysis (P <0.05). Survival was better in the more affluent states (P <0.05). Males were affected at an older age than females (P = 0.004). Blacks tended to have larger tumours although their overall survival was similar to whites. Orientals were more likely to be treated in the nineties with wide resections for smaller tumours and were located around states associated with good treatment. Orientals in Singapore and the United States had the same survival (P = 0.45). Survival in Orientals in Singapore was not significantly different from other races. The standard of healthcare for osteosarcoma varies greatly across the United States but is uniform in Singapore. Hence the observed differences in the United States were likely due to socioeconomic factors. This analysis confirms the importance of a number of prognostic variables in osteosarcoma and suggests the possibility of an ethnic and economic bias for good survival.


Ultrastructural studies in APP/PS1 mice expressing human ApoE isoforms: implications for Alzheimer's disease.
Alzheimer's disease is characterized in part by extracellular aggregation of the amyloid-β peptide in the form of diffuse and fibrillar plaques in the brain. Electron microscopy (EM) has made an important contribution in understanding of the structure of amyloid plaques in humans. Classical EM studies have revealed the architecture of the fibrillar core, characterized the progression of neuritic changes, and have identified the neurofibrillary tangles formed by paired helical filaments (PHF) in degenerating neurons. Clinical data has strongly correlated cognitive impairment in AD with the substantial synapse loss observed in these early ultrastructural studies. Animal models of AD-type brain amyloidosis have provided excellent opportunities to study amyloid and neuritic pathology in detail and establish the role of neurons and glia in plaque formation. Transgenic mice overexpressing mutant amyloid precursor protein (APP) alone with or without mutant presenilin 1 (PS1), have shown that brain amyloid plaque development and structure grossly recapitulate classical findings in humans. Transgenic APP/PS1 mice expressing human apolioprotein E isoforms also develop amyloid plaque deposition. However no ultrastructural data has been reported for these animals. Here we show results from detailed EM analysis of amyloid plaques in APP/PS1 mice expressing human isoforms of ApoE and compare these findings with EM data in other transgenic models and in human AD. Our results show that similar to other transgenic animals, APP/PS1 mice expressing human ApoE isoforms share all major cellular and subcellular degenerative features and highlight the identity of the cellular elements involved in Aβ deposition and neuronal degeneration.


Radiation resistance due to high expression of miR-21 and G2/M checkpoint arrest in breast cancer cells.
There is evidence that the extent of the G2/M arrest following irradiation is correlated with tumour cell survival and hence therapeutic success. We studied the regulation of cellular response to radiation treatment by miR-21-mediated modulation of cell cycle progression in breast cancer cells and analysed miR-21 expression in breast cancer tissue samples with long-term follow up. The miR-21 expression levels were quantified (qRT-PCR) in a panel of 86 cases of invasive breast carcinomas in relation to metastasis free survival. The cellular radiosensitivity of human breast cancer cells after irradiation was determined comparing two cell lines (T47D and MDA-MB-361) by cell proliferation and colony forming assays. The influence of miR-21 overexpression or downregulation on cell cycle progression and G2/M checkpoint arrest after irradiation was assessed by flow cytometric analysis. The expression of miR-21 was transiently increased 8 hours after irradiation in the radioresistant T47D cells and significantly changed with lower extent in radiosensitive MDA-MB-361 cells. Anti-miR-21 treated breast cancer cells failed to exhibit the DNA damage-G2 checkpoint increase after irradiation. Apoptotic activity was significantly enhanced from 7% to 27% in T47D cells and from 18% to 30% in MDA-MB-361 cells 24 hours after 5 Gy irradiation. Additionally, we characterized expression of miR-21 in invasive breast carcinomas. In comparison to non-cancerous adjacent breast tissue, tumours samples had increased miR-21 expression that inversely correlated with the distant metastases-free survival of patients (p = 0.029). Our data indicate that miR-21 expression in breast cancer cells contributes to radiation resistance by compromising cell cycle progression. These data point to the potential of combining radiotherapy with an anti-miR-21 as a potent G2/M check point inhibitor in overcoming radiation resistance of tumours.