Among scientists, there exist mixed opinions whether equine influenza viruses infect man. In this report, we summarize a 2016 systematic and comprehensive review of the English, Chinese, and Mongolian scientific literature regarding evidence for equine influenza virus infections in man. Searches of PubMed, Web of Knowledge, ProQuest, CNKI, Chongqing VIP Database, Wanfang Data and MongolMed yielded 2831 articles, of which 16 met the inclusion criteria for this review. Considering these 16 publications, there was considerable experimental and observational evidence that at least H3N8 equine influenza viruses have occasionally infected man. In this review we summarize the most salient scientific reports. Influenza A virus-like-illnesses have been recognized in horses since at least 1299, with speculation that earlier outbreaks of equid diseases could have also been due to influenza A viruses . Morens We identified 2831 articles using multiple search techniques . Six hunIn this review we found considerable historical evidence of EIV infections in man. Although they are careful to explain confounders, Morens and Taubenberger  document6.75 TCID50 equine H3N8 virus. Thirteen (86.7%) of 15 volunteers developed signs of illness, and virus was cultured from samples obtained from all volunteers at day 4 post-inoculation. Systemic illness and febrile upper respiratory illness were the most common clinical signs. The experimental data suggested that the H3N8 virus was not attenuated following passage in humans, as it was still capable of infecting and causing illness in horses.Several healthy human EIV challenge experiments were conducted in the 1960s. In 1965, hospitalized volunteers received challenges with live equine H3N8 virus (A/Equi-2/Miami/63) and were carefully monitored for evidence of infection. Five healthy adult volunteers each received 2.5 mL of undiluted equine H3N8 inoculum , among which 1 mL was administered by pipette directly into the nasal cavities and 1.5 mL nebulized into the nasal cavities and oropharynx . Each suWhile there are scattered reports in the news media and historical science writings that horses, dogs, humans, and even cats concomitantly developed ILIs ,2,29, moThe country of Mongolia has the largest horse-to-man population ratio in the world and has often suffered large epizootics of EIV . For insSoon after, at least 1400 horses were infected with H3N8 EIV during a 2007 epizootic in New South Wales and Queensland, Australia. We conducted a cross-sectional study for EIV infections among 89 people exposed to those sick horses (and 11 controls) . Serum sMore recently, we reported results from our 2005 cross-sectional study of 94 horse-exposed adults and 34 non-exposed controls from three sites in the US state of Iowa. Employing three different types of assays against two strains of H3N8 and one strain of H7N7 EIV, data suggested that at least a portion of horse-exposed adults had been previously infected with an A/equine/Ohio/2003(H3N8)-like strain. Eleven  horse-exposed adults and two  control subjects had microneutralization titers ≥1:80. Among the horse-exposed adults, 18 (19.1%) were positive by a novel neuraminidase inhibition assay and eight (8.5%) had elevated enzyme-linked-lectin assay titers ≥1:10. In the biostatistical analyses, work as an equine veterinarian was associated with increased seroreactivity.At least three H3N8 EIV human challenge studies ,18,19 clWhile the observational studies of human EIV infection in Mongolia ,21 and ADuring our review we became aware that there have been a number of non-equine influenza A viral segments identified in horses. In searching the Influenza Research Database (IRD), we found that genome segments related to influenza A H1N8, H5N1, H7N1 and H9N2 strains have all been detected among samples collected from horses. Although these identifications seem to be rare, accounting for less than 0.5% of all EIV detections, it may indicate that horses are at least susceptible to more diverse strains of influenza A virus than previously thought. For example, in 2011 a novel H9N2 influenza A virus was detected, isolated, and fully sequenced in a study of horses in Guangxi Province, China . Additionally interesting findings are that equids are not always H3N8 EIV “dead-end” hosts. H3N8 EIVs have been detected in dogs in USA, UK and Australia ,33,34,35Even with the experimental and observational data supporting evidence that H3N8 EIVs may infect man, we must be careful to temper these findings with the knowledge that serological assays against zoonotic influenza, such as equine H3N8, may be confounded by non-specific assay inhibitors and pre-existing antibodies against other influenza strains such as those caused by other human H3 viruses or vaccine.In February and March 2016, we searched English, Chinese and Mongolian literature databases, as well as queried senior scientists in Mongolia for relevant literature. English-based databases included: PubMed, Web of Knowledge and ProQuest. Chinese-based databases included: Chinese National Knowledge Infrastructure (CNKI), Chongqing VIP Database and Wanfang Data. Similarly, MongolMed was queried for related Mongolian citations. The following search query was used for PubMed, Web of Knowledge, and ProQuest databases: (“equine influenza” OR “horse influenza” OR “H3N8” OR “H7N7”) AND (“human infection” OR zoono*). The following search query was used for CNKI, Chongqing VIP Database, and Wanfang Data: SU = ‘Maliugan’ OR SU = ‘Maliuxingxingganmao’ OR SU = ‘equine influenza’ OR SU = ‘horse influenza’ OR SU = ‘H3N8’ OR SU = ‘H7N7’. The search terms “human infection” OR zoono* were not included when searching the Chinese literature databases, as the number of articles identified when using these terms was very low. Chinese papers identified with equine influenza search terms were instead manually reviewed to select papers that also included “human infection” or zoono*. We searched MongolMed for citations using “equine influenza” OR “horse influenza” OR “адууны томуу”. Mongolian influenza experts were also interviewed for their knowledge of related reports.The historical, observational, and experimental data are compelling in supporting the premise that EIV infections occasionally occur in man. While in recent years, human infections with EIVs have not often been associated with signs of infection, the propensity for influenza A viruses to change makes these viruses worthy of our attention. In particular, should H7N7 EIV strains again emerge, most humans would have little cross-reacting antibody, and the threat to humans might be quite different than that commonly seen today for H3N8. We believe these observations support close surveillance for novel influenza virus emergence among equids.


We showed an abnormal β-carotene distribution among serum lipoproteins and reduced hepatic β-carotene uptake in −/−Ldlr dams. These data strongly imply that LDLr significantly contributes to β-carotene uptake in the adult mouse liver. In contrast, LDLr does not seem to mediate acquisition of β-carotene by the placental–fetal unit.Vitamin A regulates many essential mammalian biological processes, including embryonic development. β-carotene is the main source of vitamin A in the human diet. Once ingested, it is packaged into lipoproteins, predominantly low-density lipoproteins (LDL), and transported to different sites within the body, including the liver and developing tissues, where it can either be stored or metabolized to retinoids (vitamin A and its derivatives). The molecular mechanisms of β-carotene uptake by the liver or developing tissues remain elusive. Here, we investigated the role of the LDL receptor (LDLr) in β-carotene uptake by maternal liver, placenta and embryo. We administered a single dose of β-carotene to  Vitamin A plays a pivotal role in several biological functions including the normal growth and development of the mammalian embryo  by regulIn the bloodstream of humans and a number of animal models, β-carotene has been found to be associated with all types of lipoproteins to a varying degree, with the low-density lipoprotein (LDL) being the main carrier of this carotenoid, transporting about 60%–70% of total serum β-carotene ,9,10. ApLdlr gene, we showed that LDLr participates in the uptake of β-carotene by the maternal liver but not by embryo or placenta.Given that LDLr is prominently expressed in the liver as well as in the placenta ,14,15,16Ldlr) were used . All mice, obtained from The Jackson Laboratory  were maintained and bred on a standard nutritionally complete chow diet containing 18 IU of vitamin A/g diet and 1.2 ppm of carotenoids . Genotype was confirmed by PCR according to vendor’s protocol. At approximately three months of age, females were mated according to one of the following schemes: (1) Ldlr+/− females (n = 11) mated with Ldlr−/− males; (2) Ldlr−/− females (n = 11) mated with Ldlr+/− males. In both instances, each dam carried both homozygous and heterozygous embryos. Onset of pregnancy  was established by the occurrence of a vaginal plug, the morning after the mating. Diet and water were available to the animals on an ad libitum basis. Mice were maintained on a 12:12 light/dark cycle (light on from 7 a.m. to 7 p.m.). All experiments were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals [Mice lacking either one or both copies of the LDL receptor gene  injection at mid-gestation, following a protocol previously established in our laboratory to circumvent the high mouse intestinal BCO1 cleavage activity and yield detectable intact β-carotene in the maternal bloodstream without signs of embryonic vitamin A toxicity ,19. An ed < 1.006 g/mL), LDL + IDL fraction (d = 1.023–1.06 g/mL) and HDL fraction (d = 1.063–1.21 g/mL), as previously described [d = 1.004 g/mL) and centrifuged at a speed of 70,000 g in Beckman TLA 120.1 rotor  for 4 h. The top layer containing VLDL + chylomicrons was removed using a 100 µL Hamilton glass syringe . The remaining 100 µL was mixed with 100 µL of KBr solution (d = 1.12 g/mL) and centrifuged at the same speed for 18 h. The top 100 µL of the LDL + IDL fraction and the bottom 100 µL of the HDL fraction were separated.Mouse serum was subjected to density gradient ultracentrifugation to separate VLDL + chylomicron fraction  and triglycerides , according to the manufacturer’s procedures.v/v) as the mobile phase. A Dionex Ultimate 3000 HPLC system and a computerized data analysis workstation with Chromeleon software were used. Retinol, retinyl esters and β-carotene were identified by comparing retention times and spectral data of experimental compounds with those of authentic standards. Retinyl acetate  and echinenone  were added as internal standards. Detection limits are as follows: for retinoids-serum <0.1 ng/dL and tissues <1 ng/g; for β-carotene-serum <1 ng/dL and tissues 10 ng/g).Reverse-phase HPLC analysis was performed to measure serum and tissue retinoid and β-carotene concentrations, as previously described ,21. Brie® , according to the manufacturer’s instructions. RNA was reverse transcribed to cDNA using the instructions and reagents (random hexamer primers were used) from Transcriptor First Strand cDNA Synthesis kit . A LightCycler 480 SYBR Green I Master kit  was used for qPCR. For the qPCR experiments, 300 nM of each specific primer , together with 10–50 ng of cDNA equivalent of the total RNA was used. β-actin was used as the reference gene. All samples were run in duplicates or triplicates. NRTC and the non-template were included within each experiment as controls. After enzyme activation , 35–40 PCR amplification cycles were performed: 10 s at 95 °C, 20 s at 58 °C, and 30 s at 72 °C. At the end of each run, samples were heated to 95 °C with a temperature transition rate of 0.11 °C/s to construct dissociation curves. From the instrument, cycle thresholds (Ct) were obtained for each sample for each gene of interest. To determine changes in gene expression, the ΔΔCt method was used. The expression of each gene relative to the calibrator was calculated using 2(−ΔΔCt). Gene expression changes were expressed as mRNA fold change from the control group (Ldlr+/− + vehicle). Primers and amplicon size are as previously reported [Total RNA was extracted from a sample of liver and individual placentas and embryos using TRIzolreported .Ldlr+/− and Ldlr−/− mice was loaded onto a 12% SDS-PAGE. Polyclonal rabbit anti-mouse RBP  and polyclonal rabbit anti-mouse albumin  were used for immunodetection. Albumin was used as a loading control. The quantification of the membranes was completed by densitometry analysis with Quantity One Program . Western blot analysis was repeated twice.Serum RBP levels were assessed by Western blot analysis according to standard procedures. An amount of 1 μL of serum samples (diluted 1:10) from Ldlr+/− and Ldlr−/− embryos from both Ldlr+/− and Ldlr−/− dams. Normally distributed data were analyzed by 2-way ANOVA followed by a Student’s t-test or one-way ANOVA. Data with a non-normal distribution were analyzed by a Kruskal–Wallis test, followed by a Mann–Whitney test. Analyses were performed with GraphPad Prism® version 5.0c. A p-value < 0.05 was used to establish statistical significance.The 2X2 experimental design yielded four experimental groups consisting of Ldlr+/− females with Ldlr−/− males and vice versa. This experimental design, in which each dam carried both Ldlr+/− and Ldlr−/− embryos, enabled us to evaluate the effects of the lack of placental and embryonic Ldlr in the context of a lipoprotein metabolism more or less compromised depending upon the number of Ldlr knockout alleles in the dam [Ldlr gene would impact the concentration of β-carotene and retinoids in maternal serum. As previously reported, circulating β-carotene was undetectable in vehicle-injected control dams [Ldlr+/− and Ldlr−/− groups  isolated from sera of reported , cholest females .Ldlr−/− dams showed increased TG concentrations in the VLDL + chylomicrons fraction compared to Ldlr+/− females  were elevated when compared to Ldlr+/− livers. However, β-carotene availability effaced this effect between the genotypes (Bco2 mRNA levels showed only a trend towards an increase in Ldlr+/− dams upon β-carotene administration (p = 0.06), but there was no treatment- or genotype-dependent difference in its transcription levels (Bco1) [Ldlr+/− and Ldlr−/− mice. qPCR analysis of the key regulators of tissue retinoid homeostasis revealed a significant reduction in Rdh10 mRNA in the Ldlr−/− compared to Ldlr+/− dams upon β-carotene treatment  . Thus, creatment . Moreove+/− dams , overall+/− dams , it is d+/− and Ldlr−/−) were obtained from both Ldlr+/− and Ldlr−/− dams treated with vehicle or β-carotene, resulting in eight experimental groups , the concentration of serum retinyl esters, likely associated with circulating lipoproteins, is significantly reduced, at least in the Ldlr−/− dams. These data suggest a potential competitive mechanism between β-carotene and retinyl esters for the incorporation into lipoprotein particles, at least in this model of impaired lipoprotein clearance. The mechanism of incorporation of retinoids and carotenoids into lipoprotein particles is currently unknown, although recent data from our lab strongly suggest that microsomal triglyceride transfer protein (MTP) may mediate this process in the case of β-carotene [Ldlr+/− and Ldlr−/− dams without changes in serum levels of RBP. Whether the higher serum retinol is transported within lipoproteins and is a consequence of the reduced serum retinyl ester concentrations upon β-carotene treatment remains to be established. However, this seems to be a genotype-dependent effect, as it was not previously observed in wild-type pregnant dams administered β-carotene under the same experimental protocol [Given that the majority of serum β-carotene circulates in association with LDL, in both humans and experimental animal models ,9,10,33,ype mice ,34. Theyype mice . Our stucarotene . Interesprotocol ,17.Ldlr+/− and Ldlr−/− dams. It is of note that even the absence of a single copy of the Ldlr gene reduces lipoprotein clearance [Ldlr alters β-carotene and retinoid incorporation within serum lipoproteins.In contrast to wild-type dams in which the LDL + IDL fraction contained the highest amount of β-carotene , under tlearance ,36. ThusWhether the above-mentioned altered proportion of circulating vitamin A forms in mice lacking LDLr impacts retinoid content of the liver—the main body storage site of vitamin A—is still unclear. Hepatic retinoid levels were undistinguishable between vehicle- and β-carotene-treated mice, even though some potential difference in the capacity to maintain retinoid homeostasis may exist between the two strains, based on our qPCR analysis. However, in this study, β-carotene was only administered acutely (single dose). Thus, the effects of prolonged provitamin A availability on the retinoid status of this organ remain to be established.Ldlr resulted in the ~50% reduction in hepatic β-carotene accumulation; this was unlikely due to enhanced metabolism of the provitamin A carotenoid in this organ, as assessed by qPCR analysis of the two β-carotene cleavage enzymes Bco1 and Bco2. Upon β-carotene availability, the expression levels of Bco1 were similar in the liver of Ldlr+/− and Ldlr−/− dams, suggesting that the ability to metabolize intact β-carotene by symmetric cleavage was unaffected by the complete lack of Ldlr. Nevertheless, hepatic Bco1 was upregulated in response to β-carotene availability but only in the Ldlr+/−. Although it seems reasonable that the liver may enhance its ability to convert the available β-carotene into retinoids for storage, retinoid levels were similar in the liver of Ldlr+/− vehicle- and β-carotene-injected dams [Bco1 was not observed in wild-type dams β-carotene-supplemented with a similar protocol [Bco1 expression that could also possibly explain the increased baseline expression levels of the β-carotene symmetric cleavage gene Bco1 in the Ldlr−/− livers. Further studies are needed to confirm this hypothesis and clarify the underlying molecular mechanisms. The hepatic transcription of Bco2, induced by carotenoids in vivo [Ldlr−/− dams. Our data suggest a potential upregulation of the asymmetric cleavage pathway in the liver of the Ldlr+/− dams. However, this effect unlikely contributed to the difference in hepatic β-carotene content between Ldlr+/− and Ldlr−/− dams. Together, these data point, for the first time, to a critical role of the LDL receptor in facilitating the uptake of β-carotene in the livers of adult pregnant mice. In agreement with this hypothesis, Ldlr mRNA levels increased about 30% in the liver of wild-type dams supplemented with a single dose of β-carotene (data not shown). It is also noteworthy that the hepatic uptake of circulating β-carotene is not fully abrogated in the absence of LDLr, clearly suggesting that, for example, other lipoprotein receptors such as LRP1 and VLDLr, that can take up LDL, albeit less efficiently than LDLr, may assist in cellular internalization of β-carotene by the hepatocytes. Indeed, changes in liver-specific expression of these lipoprotein receptors during gestation have previously been reported in mice lacking LDLr [Ldlr−/− mice fed an atherogenic diet. Based on our data, we could speculate that in Ldlr−/− mice, β-carotene not taken up by the liver could accumulate in the heart, among other tissues, and may contribute towards the reduction of atherogenic phenotype such as fatty streak lesions in the heart. Further studies investigating the underlying mechanism of action of β-carotene in the heart would be of great interest.Remarkably, the absence of ted dams ,38. Moreprotocol . Thus, w in vivo ,39, was ing LDLr . Interesing LDLr , a diet Ldlr genotype, disfavor a possible involvement of either maternal or placental/embryonic LDLr in regulating embryonic β-carotene acquisition and its consequent local metabolism by the cleavage enzymes. In agreement with these observations, the embryos from the dams lacking Ldlr developed normally without congenital defects or mortality associated with vitamin A deficiency or excess, under this experimental condition. Currently, we cannot predict the effects of prolonged provitamin A availability on the retinoid status of placenta and embryo.Contrary to the liver, the lack of difference in β-carotene accumulation in the placentas and embryos of the supplemented dams, as well as the similar retinoid levels, irrespective of the maternal-fetal Ldlr gene have been identified. Although severity of these mutations in causing hypercholesterolemia varies greatly, many of these lead to a several-fold elevation in the serum cholesterol [apoE genotype, especially in the liver [Ldlr and/or apoE may play a crucial role in the hepatic uptake of carotenoids incorporated into lipoproteins. In the case of vitamin A deficiency, such populations may be at a greater need of effective β-carotene supplementation strategies, which may be different from the ones utilized in the case of populations with normal lipid metabolism.We provided evidence that LDLr contributes significantly to mediate β-carotene uptake in the adult mouse liver, at least during pregnancy, whereas it does not seem to play a major role in placental and embryonic uptake of the provitamin A. In humans, several mutations (over 1000) in the lesterol ,43,44. Mlesterol . Intereshe liver . Thus, w


A review of materials and fabrication methods provides recommendations for microdevices for small-angle neutron scattering. I ≃ 10−3 cm−1). For applications requiring higher durability and/or chemical, thermal and pressure resistance, sintered or etched boron-free glass and silicon devices offer superior performance, at the expense of various fabrication requirements, and are increasingly available commercially.A comparative examination is presented of materials and approaches for the fabrication of microfluidic devices for small-angle neutron scattering (SANS). Representative inorganic glasses, metals, and polymer materials and devices are evaluated under typical SANS configurations. Performance criteria include neutron absorption, scattering background and activation, as well as spatial resolution, chemical compatibility and pressure resistance, and also cost, durability and manufacturability. Closed-face polymer photolithography between boron-free glass (or quartz) plates emerges as an attractive approach for rapidly prototyped microfluidic SANS devices, with transmissions up to ∼98% and background similar to a standard liquid cell ( A variety of flow cells   in solution. These experiments are generally implemented by continually varying input compositions and acquiring SANS data at a fixed position in a microdevice, often overilluminating several channels, in both continuous . In these experiments, a square beam of 1 × 1 cm was employed, illuminating ∼16 channels. The beam illuminates a location along the microdevice selected to ensure mixing of various inputs but limited compositional dispersion, defining a (time-varying) system composition with prescribed precision. Given the comparatively large sample volume probed, sub-second acquisition times are easily attainable, as illustrated with a micellar solution in Fig. 1b), since the effective sample volume reduction is modest. For instance, the illuminated channel area is typically 50–70% of the beam footprint (the remainder being occupied by the device matrix), and microchannel depths are typically 100–1000 µm. As a result, sample volumes are approximately 5–50% of those of a common liquid cell, requiring a modest adjustment of SANS measurement times. Rapid and precise contrast matching measurements, and systematic dilution or composition scans in ∼10 min  time scales, are readily possible , circumventing the traditional, sequential and time-consuming preparation of discrete samples, loading of cells and use of sample changers. Further, microfluidic SANS also allows adaptive scanning of parameter space  fluids. Fig. 1e) illustrates a contraction–expansion geometry for which scattering from a single ∼50 µm channel was attainable with a concentrated surfactant solution, shown in Fig. 1f), and acquisition times down to 1 s per spectrum. For instance, cross-slot (and four-roll mill) and periodic constriction geometries, shown in Figs. 1g) and 1h), can be fabricated and the effect of flow quantified, beyond the traditional flow geometries accessible, as introduced in Table 1e.g. parabolic or plug flow) within the microchannel, which normally requires ancillary flow measurements  of 1–500 nm. Experiments measure the differential macroscopic scattering cross section of the sample, a product of the form factor P(q) and the structure factor S(q):N/V is the number density, Vp is the volume of a solute particle, (Δρ)2 is a contrast factor and B is the scattering ‘background’. P(q) contains information about the solute’s shape and size and S(q) about the spatial correlations between individual scattering species. Δρ can be effectively tuned by selective deuteration, enabling the direct determination of P(q) in concentrated or inter-penetrating systems. Given the number of excellent introductions to SANS and data analysis, we concentrate here on microdevice fabrication aspects.SANS probes spatial correlations between ensembles of atomic nuclei on the nanoscale, typically covering a wavenumber range of 0.001 3.1.q range relevant for SANS (approximately 0.001–0.5 Å−1) compared with the samples of interest; (iii) low neutron activation and/or short half-life decay; (iv) chemical compatibility with the sample and ability to withstand the required pressure, typically up to 1–10 bar , without deformation, as required for absolute SANS intensity calibration; (v) feature resolution in the range ∼1–1000 µm; and (vi) compatibility with rapid prototyping techniques  if flexible channel design is necessary. Specific applications may have additional requirements, such as oxygen permeability , surface functionality for prescribed wetting in multiphase flows, or control wall slip.Microdevices for SANS experiments should meet a number of requirements. In short, they must exhibit (i) low neutron absorption; (ii) weak scattering in the experimental wavenumber 3.2.−1) must take into account the various contributions to scattering, transmission and absorption from the microdevice components. A detailed description of the data reduction and calibration steps, including useful simplifications and assumptions, is presented in the supporting information.Fig. 2d to width w  should satisfy 4W/Dw/d, where D is the sample-to-detector distance and W is the maximum distance between the beam centre and the edge of the detector. For a typical high-q setup , d/w greater than 1–3 should be used. A full offset of the detector (often used to enlarge the q window) can increase this requirement by a factor of 2.At high scattering angles, additional geometric data corrections may be necessary since neutrons scattered close to the edge of a channel will pass through the channel matrix before reaching the detector, and are thus attenuated or scattered in a non-trivial manner. As a rule of thumb, the ratio of channel depth e.g. using MgF2 lenses) generally focus the beam at the detector to access lower q values, as required for very small angle scattering, vSANS; focusing at the sample is not suitable as a strategy to decrease the beam size for microfluidics owing to the accompanying increase in beam divergence, which in turn restricts the accessible low-q range  are employed to maximize the illuminated area and thus the scattering signal. Reducing the diameter of the diaphragm, and/or altering its shape, is thus needed to accommodate small samples. Evidently, reducing the beam footprint also reduces the neutron flux, in near proportionality to the area. For instance, reducing a circular beam of 12 mm diameter to 100 µm decreases the neutron flux by ∼106, limiting the feasibility of some flow-mapping experiments, such as those shown in Fig 1e). Generally, enriched hot-pressed 10B4C is preferred as a manufacturing material as it contains no hydrogen and produces many fewer γ-rays than the Cd or Gd counterparts. These apertures are usually 2 mm thick and cut by electro-erosion, as 10B4C is an extremely hard and brittle material. Slit apertures as small as 50 µm can be made to increase the spatial resolution and enable fine scanning. Furthermore, rectangular slits or multi-slit masks (registered with microchannels) can be desirable to increase the illuminated area and thus the signal and to minimize background scattering, although this requires precise alignment and positional stability.Neutron focusing optics  4 m distance, resulting in a q range of 0.009 q−1.Neutron scattering measurements were performed on beamline D22 at the ILL reactor  and on the LOQ beamline at the ISIS pulsed source . On D22, the wavelength was fixed at a representative λ = 6 Å, and three sample-to-detector distances were used , yielding a 4.2.t ≃ 140 µm) and soda lime glass slides (t ≃ 1 mm) were purchased from Fisher. Quartz slides (t ≃ 0.44 mm) and fused silica slides (t ≃ 1.1 mm) were purchased from H. Baumbach and Co. Ltd. Silicon wafers  were purchased from Si-Mat (Germany). A poly(dimethylsiloxane) (PDMS) film (t ≃ 0.6 mm) was produced by mixing Sylgard 184 elastomer with a curing agent (both obtained from Dow Corning) in a 10:1 weight ratio. The film was then degassed and cured at 348 K for 1 h. Hydrogenated and deuterated polystyrene (PS) and hydrogenated poly(methyl methacrylate) (PMMA) films (t ≃ 1 mm) were pressed from standard grades. A thiolene film (t ≃ 0.54 mm) was prepared by photocuring the optical adhesive Norland NOA-81 (purchased from Edmund Optics) with UV light. Kapton film (t ≃ 0.09 mm) was obtained from Müller Ahlhorn (Germany). An aluminium slab (t ≃ 1.2 mm) was obtained from the Imperial College London workshop; its scattering pattern was close to that of commercial (∼4 µm thick) aluminium foil.Borosilicate cover slips . Most low atomic number (Zdq* = 2π/d), should be avoided. Polycrystalline materials will also scatter in the low-q region owing to grain boundary interfaces . Otherwise, a range of elements and materials would appear to yield reasonably high-quality SANS cells , exhibits a neutron transmission of ∼96% (at λ = 6 Å) and a scattering intensity of ∼5 × 105.2.supporting information. In addition, the thickness required for microfabrication must be considered and thus we also estimate (bottom row) the transmission ranges of corresponding devices. For the different glasses, we assume the device thicknesses of our fabricated devices described in §6et al., 2012et al., 2009t = 250 µm for thiolene  windows are thus required. Polymers show a transmission of ∼0.6 ± 0.1 for 1 mm thick slabs, which translates to device transmissions of ∼0.8. Kapton devices with thin walls  or titanium will typically decay to safe levels within approximately 1 h, and the same mass of Si or SiO2 (e.g. quartz or fused silica) requires approximately 1 d after a reference 10 h exposure at 108 neutrons cm−2 s−1. By contrast, Cu would require a few days, Zn and Fe a few years, and Ni and Co decades, under identical conditions  to 10 mm (ρ ≃ 1 g cm−3) and a beam footprint of 1 cm2. Evidently, sufficiently thin metal sheet windows with a low mass (if compatible with microfabrication) and smaller beams will eventually yield lower or even imperceptible radioactivity. A configuration with a beam area of 0.25–1 mm2 impinging upon two metal windows of 10–50 µm thickness , which yields an acceptable decay time for a range of metals, as illustrated in Table 2Prolonged exposure to neutrons may induce considerable radioactivity in materials, which must decay to safe levels to enable handling, for instance when changing samples or upon completion of an experiment. Low activation and/or a short decay half-life are thus desirable for a microdevice material. While elements H, C, N and O, constituents of polymeric materials, are intrinsically safe, 1 g of Al, sapphire (Ale.g. flow stabilization or cleaning) to minimize neutron exposure.The above estimates for 10 h exposure, albeit much longer than a single-spectrum acquisition, are reasonable for a series of microfluidic SANS experiments. While the beam is generally open throughout a SANS experiment, it is recommended that the beam is shut off during down-time  is shown in the top graphs. As mentioned above, this can be somewhat misleading, as we are interested in the scattering of the microdevice compared with that of a sample (confined to a given channel depth). The lower panels thus show the same data multiplied by the likely device thickness tD. The scattering signal for all glass and silicon specimens considered is weak compared with that of samples ‘compatible’ with microfluidic SANS, which should scatter above ∼1 cm−1 to enable reasonable acquisition times. Borosilicate exhibits higher scattering (in addition to lower device transmission) and is thus inferior to other glasses. By contrast, silicon is outstanding in this q range and thus even thick devices (of the order of centimetres) would be well suited for SANS. In general, neutron activation is not of concern, provided that the window thicknesses are kept to a minimum (tens to hundreds of micrometres).Fig. 45.4.2.i.e. device fabrication within hours of the initial design  owing to their high hydrogen content. The absolute scattering profiles of representative polymers are shown in Fig. 4b), accompanied by their figures of merit ItD in the panel below, which are consistently above that of a Hellma cell. Polymers are generally classified into elastomers, thermosets and thermoplastics.Polymer-based microdevices are generally inexpensive compared with glass and silicon devices and are amenable to rapid prototyping, e.g. in biology. On the other hand, PDMS suffers from poor solvent compatibility as it is swollen by common organic solvents  and high failure strain. By far the most commonly used elastomer in microfluidics is poly(dimethylsiloxane), PDMS  and microdevices can be fabricated by moulding under pressure, or shaped by other techniques including laser ablation. PMMA and cyclo-olefin-copolymer (COC) devices with wall thicknesses of 130–250 µm, and polyimide (Kapton) devices with a thickness of 75 µm, have been fabricated by hot embossing and laser ablation, respectively, and used in SAXS experiments  shows the scattering of five selected polymer systems: hydrogenated PS, PDMS, Kapton and (cross-linked) thiolene, in line with I(PMMA) ≃ 0.6 cm−1 and I(polyethylene) ≃ 0.62 cm−1  and low availability may make this unfeasible. Deuteration yields an incoherent cross section ∼15 times lower that of hydrogenous polymers, resulting in potentially excellent window materials, in particular at high q, as shown by the d-PS profile. Hydrogenous polymeric devices therefore perform comparatively poorly for use in microfluidic SANS cells, unless thin-walled polymeric devices are used, provided that they are mechanically robust and enable robust inlet/outlet connections. Taking an illustrative channel depth of 500 µm, a device (top and bottom) wall thicknesses of ∼25 µm would be needed for the channels to scatter similarly to D2O . However, a number of ‘lab-on-a-foil’ polymer devices with windows down to 25 µm have been demonstrated in SAXS , can scatter strongly in the high-5.4.3.et al., 2011et al., 2012et al., 2001et al., 2003et al., 2008et al., 2008c) shows the scattering profiles for nickel, aluminium, vanadium and steel, with the expected performance in a device shown by ItD in the row below. The ability to fabricate mechanically robust thin metal devices leads to generally good neutron cells, in particular in the range q−1. For example, nickel devices with 10 µm thick walls , although its quality decreases at lower q values, reaching the value of D2O at q ≃ 0.02 Å−1. However, we note the limited chemical stability of aluminium, and its susceptibility to solutions of varying salt and pH can impact the surface roughness and composition of the microchannels.Metals offer high thermal and electric conductivity, and generally have high Young’s modulus, tensile strength and melting point. Gold, nickel and copper are the most commonly used metals in microfabrication  and low incoherent and background scattering (I ≃ 0.0002–0.005 cm−1).Ceramics are inorganic, hard insulating materials with high melting temperatures and excellent chemical resistance, which have been employed recently for microfabrication  devices are amenable to rapid prototyping in most university laboratories, are inexpensive, and enable iterations and refinements, which are particularly important for flow-SANS experiments. By contrast, microfabrication in glass or metal, for instance, generally offers more durable devices, able to sustain higher pressures and attractive for repeated or prolonged use, such as in phase mapping. Key features of selected materials and approaches are summarized in Table 36.1.Given the low background scattering of (boron-free) glass and quartz and the ease of fabrication of photopolymerization, an attractive fabrication approach consists of so-called ‘closed-face’ lithography, illustrated in Fig. 5et al.  or quartz plates. Panel II depicts a longer procedure, yielding devices compatible with both SANS and SAXS. The procedure is as follows. (a) Small holes are drilled in a thick (1 mm) glass slide which serve as inlet(s) and outlet, and a large hole/window is drilled around the region for observation with neutrons. We find that drilling the inlet and outlet holes before the large central one significantly reduces breakages while drilling. This slide will act as a ‘reinforcement’ for the thinner glass slides (e.g. t = 140 µm borosilicate glass slides). A conventional high-speed drill with a diamond drill bit can be employed. (b) A few drops of thiolene (NOA81) are cast onto a thin cover slide and the reinforcement slide is sealed under UV light, any excess adhesive being removed with a razor blade. (c) Holes are drilled through the thin cover slide on the inlet(s) and outlet positions; the thin and thick slides are then bonded together prior to device patterning. (d) To define a precise channel depth, spacers are placed on top of another thin cover slide, thiolene is poured on top and the reinforced slide sandwiches the thiolene resist layer. (e) The device is flipped, a negative photomask (i.e. with microchannels in black) is placed over the assembly and the whole is exposed to a prescribed UV dose . (f) The liquid thiolene that remains under the masked sections is flushed out using ethanol, acetone and compressed air. (g) The device is post cured by exposure to high-intensity UV for at least 30 min. (h) Nanoports  are sealed to the device using fast-curing Araldite glue, and the device is ready for operation. The entire process takes approximately 1 h, yielding devices suitable for SANS . Borosilicate glass has a similarly low background but much lower transmission, and suitable devices must therefore have thin walls (∼2 × 140 µm for 80% transmission).6.2.et al., 1998et al., 2016d). After the channels have been developed, a replicated PDMS slab  and 6f). Finally, in step (g) tubing is connected to the PDMS module onto pre-cored ports, and an outlet connector is attached through the reinforcement slide.While the fabrication of PDMS devices by SU-8 mould replication is well established  for high-aspect-ratio channels. Access ports for inlets or outlets can also be wet etched, as well as drilled or sand blasted. Alternative processes couple laser ablation with chemical etching . Finally, the patterned surface is bonded by direct bonding or fusion bonding at high (∼923 K) temperatures or with an adhesive, before the fluidic ports are connected. Commercial suppliers often make available robust mounting platforms with push-fit tubing or ferrule connectors for additional robustness (e.g. against leaks or detachment). Typically, these devices can withstand pressures in excess of 100 bar . While borosilicate glass is generally unsuitable because of its high absorption (see Table S1 in the supporting information), a common and inexpensive crown glass  with a low boron content is well suited, as demonstrated in previous contrast-variation and phase-mapping experiments .All-glass devices can offer superior performance compared with methods 1 and 2 in terms of durability and chemical and pressure resistance, which may warrant the higher material and processing costs required. While glass micromachining, etching and bonding . These typically employ ultrafast pulsed lasers and focusing optics or two-photon processes to pattern directly inside the glass matrix . The spatial resolution can be as high as 1–2 µm and the high optical transparency, chemical, thermal and pressure resistance, and neutron transmission make these approaches particularly suitable for SANS device fabrication.Selective laser-induced etching methods  are eme al. 2016e. These6.4.Table 3e.g. borosilicate) slides are also compatible with SAXS .Methods 1 and 2 proposed here aim to combine the high neutron quality of glass or quartz with the versatility and speed offered by polymer microfabrication. These yield high-quality neutron cells that can be manufactured within hours of initial design, albeit suffering from limited resolution . Their low fabrication cost (∼10 USD) and time (of the order of hours) mean that they are effectively disposable and device cleaning might thus not be needed . Devices fabricated by method 3 using thin . A number of glass etching and bonding techniques, as well as buried laser etching techniques, provide durable solvent-compatible optically transparent high-pressure microdevices that are particularly well suited for microfluidic SANS, in particular for applications requiring prolonged or repeated use (e.g. phase mapping). The combination of both approaches, in addition to continued advances in the three-dimensional printing of polymers and glasses, is likely to meet most requirements for microfluidic SANS.We have reviewed materials and approaches for the fabrication of microfluidic devices compatible with SANS. A few closed-face photopolymerization methods between quartz (or boron-free glass) plates emerge as effective rapid prototyping approaches, requiring facilities available in most chemical laboratories, and enable rapid iterations that are advantageous for some SANS experiments , mid q (0.01–0.2 Å−1) and low q (0.005–0.05 Å−1). Data acquisition times generally increase towards lower q ranges owing to a combination of a smaller solid angle and/or lower flux for long-wavelength neutrons, making high- and mid-q configurations particularly suitable for microfluidic SANS. Considering the inverse proportionality between acquisition time and both the absolute scattering intensity and (illuminated) sample volume, the experimental feasibility is indicated by colour in Fig. 8f). For channel overillumination, and beams of the order of centimetres, a large range of systems are compatible with microfluidic phase-mapping SANS. As a rule of thumb, the channel matrix (i.e. the spacing between channels) is 20–50% of the channel width, channel depths are sub-millimetre and counting times need to be adjusted accordingly with respect to the bulk sample. Further, operational considerations of the required mixing times and flow dispersion, relevant for the precise formulation of mixtures, must be carefully considered  or to unlock the molecular or mesoscopic underpinnings of complex fluid rheology and processing, and the mechanisms and kinetics of phase and conformation transition kinetics, amongst others. This review provides clear recommendations for choice of materials and fabrication methods for microfluidic SANS.Overall, microfluidics offers unprecedented opportunities to expedite traditional SANS experiments (8.et al. (2007et al. (2000et al. (2015et al. (1999et al. (1989For further literature related to the supporting information, see Brûlet  al. 2007, Dolník  al. 2000, Glinka  al. 2000, Hammoud al. 2000, James & al. 2000, Lide (2 al. 2000, NIST (2 al. 2000, Osman e al. 2015, Prince  al. 1999, Root et al. 1989 and Sear al. 1989.10.1107/S1600576718007264/aj5307sup1.pdfTheoretical background and additional tables and figures. DOI:


Our objectives were to elaborate nutrient budgets and to diagnose soil, foliar, and fruit nutrient balances in guava orchards fertilized with guava waste. We hypothesized that (1) guava waste are balanced fertilizer sources that can sustain crop yield and soil nutrient stocks, and (2) guava agroecosystems remain productive within narrow ranges of nutrient balances. A 6-year experiment was conducted in 8-year old guava orchard applying 0–9–18–27–36 Mg ha−1 guava waste (dry mass basis) and the locally recommended mineral fertilization. Nutrient budgets were compiled as balance sheets. Foliar and fruit nutrient balances were computed as isometric log ratios to avoid data redundancy or resonance due to nutrient interactions and the closure to measurement unit. The N, P, and several other nutrients were applied in excess of crop removal while K was in deficit whatever the guava waste treatment. The foliar diagnostic accuracy reached 93% using isometric log ratios and knn classification, generating reliable foliar nutrient and concentration ranges at high yield level. The plant mined the soil K reserves without any significant effect on fruit yield and foliar nutrient balances involving K. High guava productivity can be reached at lower soil test K and P values than thought before. Parsimonious dosage of fresh guava waste should be supplemented with mineral K fertilizers to recycle guava waste sustainably in guava agroecosystems. Brazilian growers can benefit from this research by lowering soil test P and K threshold values to avoid over-fertilization and using fresh guava waste supplemented with mineral fertilizers, especially K. Because yield was negatively correlated with fruit acidity and Brix index, balanced plant nutrition and fertilization diagnosis will have to consider not only fruit yield targets but also fruit quality to meet requirements for guava processing.The Brazilian guava processing industry generates 5.5 M Mg guava waste year Psidium guajava) is a tropical tree grown on Brazilian Oxisols and Ultisols and reaching high productivity 3 years after establishment  guava waste are balanced fertilizer sources that can sustain crop yield and soil nutrient stocks under local climate conditions, and (2) guava agroecosystems are productive within narrow ranges of nutrient balances. To verify hypothesis (1), nutrient budgets were elaborated across several fertilization treatments during six guava cropping cycles where annual climate conditions and soil stocks varied. Hypothesis (2) was verified using locally derived foliar nutrient balance standards for “Paluma” guava at high yield level under the −1 plant−1 using micro-sprinklers whenever the tensiometer installed 0.20 m below surface indicated that soil water content reached 60% of field capacity.Perennial fruit-bearing plants such as guava require conducting long-term experiments to document nutrient issues due to the continuous nutrient supply and removal and to internal nutrient cycling  to facilitate handling, and stored in the shelter. Fresh residues were collected yearly from the same process and transported to the field; the 3–5 cm thick layers were exposed to relatively fast air drying (within 1–2 weeks).2 per tree) for a total of 140 trees for the 4900 m2 experimental orchard. There were five doses of waste  applied as dried-and-ground material to facilitate manual handling, a dosage of 18 Mg dry waste-equivalent ha−1 year−1 of fresh waste as would be machine-applied in practice, and a standard treatment of locally recommended mineral fertilization. Mineral fertilizers and guava waste were spread manually each year the same day within crown projection area. The sources of mineral fertilizers were urea (45% N), ordinary superphosphate , and KCl , supplying 229 kg N ha−1, 12.5 kg P ha−1, 71.4 kg K ha−1, 31.5 kg Ca ha−1, and 19 kg S ha−1. Treatments started in March 2006 and continued yearly in January after soil sampling . The N was determined by micro-Kjeldahl including nitrate. The P and S were quantified by colorimetry and cations  by atomic absorption spectrophotometry after digestion in a mixture of nitric and perchloric acids. Boron was determined by colorimetry after ashing waste for 3 h in a muffle furnace at 550°C. The average nutrient composition of guava waste presented in Table −1 provided 202–237 kg N ha−1 and that of fresh guava waste, 289–310 kg N ha−1.Moisture content in guava waste was determined after drying at 67 ± 2°C in a forced air oven before chemical analysis  contained 260 g clay kg−1, 60 g silt kg−1, 100 g very fine sand kg−1, 390 g fine sand kg−1, 180 g medium sand kg−1, and 10 g coarse sand kg−1. Soil available nutrient levels and pH were determined yearly each December in the 0–0.20 m and 0.20–0.40 m layers where the bulk of guava roots is located , organic matter content, K, Ca, Mg, and (H + Al) according to van Raij et al. , quantified by flame photometry (K) or atomic absorption spectrophotometry , and reported as mmolc dm−3. Potential acidity (H+Al) was quantified by the SMP pH buffer method  dm−3. Cation exchange capacity (CEC) was calculated as the sum of cationic species. Base saturation was computed as the sum of molar concentrations of K, Ca, and Mg divided by CEC, to conduct BCSR diagnosis. Other nutrients were diagnosed in isolation using SLAN.The soil was classified as dystrophic red-yellow Ultisol EMBRAPA, . Soil suj et al. . The Cu,−1) on dry mass basis by nutrient content in kg Mg−1 dry mass. Nutrient removal was quantified by multiplying fruit yield in Mg fruit ha−1 (mean of 11% dry matter in fruit at harvest) by fruit nutrient concentration in kg nutrient Mg−1. Because fruit analysis was conducted at three occasions only, nutrient concentrations by treatment  were obtained averaging the ilr values then transforming them back to concentrations. Starting 90–100 days after fruit set, guava fruits were harvested 1–3 times per week during 2 months from three representative central trees per plot at proper fruit ripening, commercial size, yellow skin, and characteristic aroma for industrial processing ] where one proportion “x” is expressed relatively to its complement “(1 − x).” The closing element (1 − x) to 100% may be defined as a filling value (Fv) computed as the difference between the unit or scale of measurement and the analyzed nutrient, generally expressed on a dry mass basis. After statistical analysis, the x and (1 − x) centroids are recovered by back transforming the mean of the logistic variable to original units.The simplest function to remove one degree of freedom without losing any information from a composition closed to 100% is the logistic variable, i.e., X and Y are often reduced to a single one as dual log ratio reflecting the interaction between X and Y. Using Fv as basis, the log of X/Fv, and Y/Fv are called additive log ratios or alr , there are two alrs in the compositional vector, hence removing one degree of freedom. Again, results of statistical analysis can be back-transformed to original units from three equations: the values of log[X/Fv]and log[X/Fv], and [X+Y+Fv = 100%]. However, the alrs are not independent  from one another: the angle of 60° between alrs makes it difficult to project them into the Euclidean space  in which nutrients labeled “+1” (group numerator) are contrasted with nutrients labeled “-1” (group denominator) in each row. A part labeled “0” is excluded. The composition is partitioned sequentially at every ordered row into two contrasts until the (+1) and (−1) subsets each contain a single part. Balances are computed as follows  are geometric means of components in the + and – groups, respectively. Coefficient g!/2D−1 possible sequential binary partitions (SBP) for the D - 1 balances derivable from D-parts compositions  classification algorithm, appropriate for the Euclidean space of ilrs, as the “kknn” package launched with the R “caret” package /(TN+TP+FN+FP). To compute critical concentration ranges, we generated a large number  of uniformly distributed values between the ranges of the confidence intervals (P = 0.05) about the TN ilrs. We then back-transformed each row of randomly generated ilrs to concentrations, and finally extracted the minimum and maximum univariate concentrations across the TN ilr confidence ranges.We discretized yield between low and high yielders in order to predict the yield class using a ge Kuhn, . We optige Kuhn, . The TN ge Kuhn, . Accurac−1. Ordinary superphosphate provided Ca and S in mineral fertilization in amounts comparable to or higher than guava waste, but Mg and micronutrients were in deficit with the mineral fertilization (RM) and the control, hence potentially affecting soil Mg stocks. The pH increased from 5.0 in 2006 to 5.4–6.2 in 2010 in the 0–20 cm layer but was stationary in the 0.2–0.4 m layer, indicating reduction of potential aluminum toxicity  in simple superphosphate. Soil test SO4-S did not vary across guava waste treatments despite additions of organic S. Soil tests Cu and Mn increased significantly due likely to fungicide applications while B decreased significantly in the control treatment only. Soil tests Ca and Mg appeared low, leading to base saturation near the lower limit suggested for guava  about ilrs and the corresponding minimum and maximum concentration values for TN and TP specimens are presented in the mobile design in Figure −1, 2.81 g S kg−1, 9.00 g Ca kg−1, and 30 mg B kg−1. Because other ranges overlapped between TN and TP specimens, apparent critical values could not be computed for that guava agroecosystem.The accuracy of the partition using the k nearest neighbors (knn) classification across ilrs was 82% in cross-validation and 93% on the whole data set, with 96 true negatives, 85 true positives, 4 false negatives, and 10 false positives . Confidence intervals  could be supplemented by mineral K fertilizers to maintain soil test K above 0.7 mmolc K dm−3 at productivity levels of 60–61 Mg ha−1 or 1.0–1.2 mmolc K dm−3 at productivity levels of 68–77 Mg ha−1  ratio ranged from 11.6 and 12.2 for TN specimens and 10.4–11.3 for TP specimens due apparently to excessive P levels in TP specimens. The lower critical value for the N/P ratio was 11.45. Other estimates of N/P ranges for guava at high yield level in Brazil were 11.5–13.1  revisiting optimum soil test K and P thresholds as well as the SLAN and BCSR interpretation models used in Brazil to achieve parsimonious nutrient management in guava agroecosystems where guava waste is recycled, and (2) monitoring the nutrient balance of fruit and foliar tissues to reach balanced fertilization in relation with targeted fruit yield and quality standards, and variations in climate conditions.HS: Field work, data collection and acquisition, statistical analyses and interpretation, literature review. SP: Data modeling , graphics and tables, co-writer of the paper. DR, DA, VM: Field work, data collection, statistical analyses, and data set organization. WN: Conception and design of the experiment, laboratory methods, relation with the industrial partner, data interpretation, paper review, literature review. LP: Data set, literature review, co-writer of the paper.We are grateful to Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq), Indústria de Polpas e Conservas VAL Ltda., and the Natural Sciences and Engineering Research Council of Canada (NSERC-DG 2254) for financial support.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


P = .048), stent thrombosis , target lesion revascularization , target vessel failure , target lesion failure , myocardial infarction , and major adverse cardiac events  at 2-year follow-up. However, the incidence of target vessel revascularization  was similar between DB and non-DB patients. Patients with DB , older than 80 years  with hypercholesterolemia , serum creatinine >177 μmol/L , a history of cerebral vascular accident , or a history of myocardial infarction  were more likely to experience adverse events. In China, DM could also be served as an independent predictor of adverse outcomes after DES implantation. These patients should be reexamined more frequently.Diabetes mellitus (DM) has been proved to be a predictor of adverse outcomes after percutaneous coronary intervention (PCI). Drug-eluting stents (DESs) could reduce the adverse events in DM patients. In this study, we aimed to analyze the clinical outcome after DES implantation in diabetic versus nondiabetic patients in China. Totally, 200 Chinese DM patients and 400 Chinese non-DM patients were enrolled in this retrospective study. Compared with non-DM patients, DM patients were more likely to have a higher incidence of cardiac death (3.5% vs. 1.0%,  The most common therapy for coronary artery disease patients is percutaneous coronary intervention (PCI).More than 347 million individuals were affected by diabetes mellitus (DM) globally in the year 2008.–11 Recently, drug-eluting stents (DESs) have been proved to reduce the incidence of restenosis and the need for repeat revascularization when compared with bare-metal stents in patients with DM.,13 However, compared with non-DM patients, DM patients still have a high risk of major adverse cardiac events (MACEs) with DES.–16 Some studies found that the incidence of cardiac death between patients with or without DM is similar. But some other large sample clinical trials found that patients with DM still had a higher risk of MACE even with DES.–20 All studies above are not reported in China.DM has been proved to be a predictor of adverse outcomes after PCI including restenosis, repeat revascularization, myocardial infarction (MI), and mortality. This may be important in the etiology of thrombotic events. In Chinese patients, coagulation state is not that high, and little data are known about the outcomes of DES usage in Chinese patients.Thrombotic and thrombolytic status is different between western people and Asians. Compared with patients in western countries, platelets and thrombolysis are both inhibited in Asian patients.Thus, we conducted this study to investigate the efficiency and safety of DES in Chinese patients with or without DM. Furthermore, we also conducted a meta-analysis to systematically review the effect of DM in patients undergoing PCI with DES.22.1This study was approved by the institutional review board of Anhui Medical University, and it was conducted in accordance with the Declaration of Helsinki and internationally accepted ethical guidelines. All patients signed an informed consent form.2.23 or a WBC count <3000 cells/mm3.Totally, 600 Chinese patients treated with PCI and with DES were continuously enrolled in this retrospective study between 2010 and 2013 . Patients included in this study should meet the following criteria: age 18 to 80 years; clinically diagnosed as having chronic coronary artery disease (CAD) or acute coronary syndrome (ACS); >1 target lesion; target lesion stenosis >50%; using at least 1 DES. Exclusion criteria were: history of stroke or transient ischemic attack within 6 months; history of coagulopathy; platelet count <100,000 or >700,000 cells/mmFor patients with stable CAD, those experiencing the following indications were eligible for PCI: target lesion stenosis ≥70%; target lesion stenosis is <90% but fractional flow reserve <0.8. For patients with non–ST-segment elevation ACS, patients experiencing the following indications were eligible for PCI: unstable hemodynamics patients; refractory angina pectoris; patients with life-threatening arrhythmia; acute heart failure; with increased serum troponin levels. PCI was conducted as soon as possible when patients experienced ST-segment elevation myocardial infarction.2.3The following information was collected for patients included in our study: age, sex, height, weight, blood pressure, blood glucose level, blood lipid levels, serum creatinine levels, electrocardiography, and disease history . Quantitative coronary angiography was performed using the Philips quantitative coronary analysis system  and was assessed by an interventional cardiologist. A successful procedure was defined as <25% residual stenosis after PCI.PCI was performed according to current standard procedural guidelines.Before or during the PCI procedure, all patients received at least 300 mg of aspirin and a 300- to 600-mg loading dose of clopidogrel. After the PCI procedure, all patients were given 100 mg/day of aspirin continuously and 75 mg/day clopidogrel for at least 12 months.2.4 DM patients were further stratified by DM treatment into insulin-treated DM (ITDM) and noninsulin-treated DM (NITDM). For NITDM patients, treatments were oral glucose-lowering drugs or without medication, such as lifestyle modification.DM was diagnosed as an abnormal blood glucose level (=126�mg/dL) after an overnight fast, an abnormal glycosylated hemoglobin test (=6.5%), or an abnormal glucose-tolerance test (2 hours =200�mg/dL).2.5 TLF is composed of cardiac death, target vessel-related MI, and TLR. And TVF is composed of cardiac death, target vessel-related MI, and TVR. MACEs were defined as cardiac death plus stent thrombus plus TVR.The following information was collected at 2-year follow-up: death (cardiac death and death of other reasons), stent thrombus (at 1 month/1 year/2 year), target vessel failure (TVF), target lesion failure (TLF), target vessel revascularization (TVR), target lesion revascularization (TLR), MI, and MACE. Endpoint events were defined as death. Stent thrombus were assessed according to the Academic Research Consortium definition.2.6P value <.05 was defined as the threshold of statistical significance. For normally distributed data, mean ± standard deviation (SD) was used for statistics. And asymmetrically distributed data were expressed as median (range). Independent-sample t tests were used to calculate the differences between the 2 therapies. The Kaplan-Meier method was used to evaluate the incidence of cumulated events. Univariable and multivariable Cox proportional hazard regression models were used to compare time-dependent dichotomous events among groups.SPSS 20.0  was used for statistical analysis and 2.7MEDLINE, EMBASE, Science Citation Index, the Cochrane Library were systematically searched using the following Keywords: drug eluting stent or DES; diabetes mellitus or DM; percutaneous coronary intervention or PCI.I2. Homogeneity between trials was assessed using the χ2 test with the significance threshold set at P > .1.RevMan 5.2.6 (Cochrane Collaboration) were used for meta-analysis. Mantel-Haenszel risk ratios (RRs) with corresponding 95% confidence intervals (CIs) were calculated for the pooled outcomes. Heterogeneity was assessed by calculating 33.1Totally, 600 patients including 200 DM  and 400 non-DM patients treated with PCI were enrolled in our study. Compared with non-DM patients, DM patients had higher body mass index (BMI), and were more likely to have a history of hypertension, hypercholesterolemia, peripheral vascular disease, kidney function deficiency (serum creatinine >177 μmol/L), and family history of coronary disease. In DM patients, all baseline characters were similar between ITDM and NITDM, except that ITDM patients were more likely to have kidney function deficiency (serum creatinine >177 μmol/L) , and the difference between ITDM and NITDM patients group (P = .180) or between NITDM and non-DM patients group (P = .260) was not significant . Together, 11 patients  experienced cardiac death. The incidence of cardiac death was significantly lower in non-DM patients group (1.0%) compared with DM patients (3.5%). But the difference was similar between ITDM (5.9%) and NITDM patients (2.2%) (Table 3.3.2P = .040). The difference between DM and non-DM patients or ITDM and NITDM patients at follow-up of 1 month, 1 month to 1 year, and 1 year to 2 year was of no significance (Table P < .001) or NITDM patients (P = .030). However, the difference between NITDM and non-DM patients group (P = .730) was not significant  and 2 non-DM patients were found to have stent thrombus (ce Table . Cumulat3.3.3P < .001). Nevertheless, non-DM patients (n = 17) also experienced significantly less incidence of TLF compared with DM patients  (P < .001) , non-DM patients n = 32) experienced significantly less incidence of TVF (2 experie3.3.4P = .340). Compared with non-DM patients (n = 7), more DM patients  experienced TLR (P = .005). Both the differences of TVR and TLR between ITDM and NITDM patients were not significant  and 22 non-DM patients P = .40. CompaP = .040) or NITDM patients group (P = .009). The difference between ITDM and NITDM group (P = .980) was not significant. Cumulative incidence of TVR was significantly higher in ITDM patients when compared with non-DM patients (P = .040) or NITDM patients (P = .030). However, the difference between NITDM and non-DM patients (P = .440) was not significant . The difference between ITDM and NITDM patients was not significant (Table P < .001) or NITDM patients (P = .030). However, the difference between NITDM and non-DM group (P = .550) was not significant  than DM patients  (3.3.6P = .001). Moreover, NITDM patients experienced lower incidence of MACE than ITDM patients (P = .040) (Table P < .001) or NITDM patients (P = .040). Moreover, the difference between NITDM and non-DM group (P = .040) was also significant  patients experienced significantly lower incidence of MACE than DM patients  , with hypercholesterolemia , serum creatinine >177 μmol/L , a history of cerebral vascular accident , or with a history of MI  were more likely to experience adverse events , the authors found that 22 studies reported the effect of DES both in DM and non-DM patients.3.4.1,29,33,34 and the data in the present study reported cardiac death; the pooled analysis showed that DM patients had a higher incidence of cardiac death   . Eleven studies,26,28,33 as well as our data reported TVR; the pooled analysis showed that DM patients had a higher incidence of TVR    . This may be attributed to their population characteristics. In their study, we found patients were more likely to have hyperlipidemia (81.3%), more history of chronic renal insufficiency (12.8%) and more history of MI (73.2%), and all these factors were independent risk factors which were associated with cardiac death in our study. Thus, the death rate would be higher in their study.In our study, we found that patients with DM experienced more cardiac death (3.5% vs. 1.0%). Our result was in accordance with other studies .We also found that a higher incidence of adverse events could be observed in DM patients . We also conducted a meta-analysis to figure out the relation between DM and adverse events. From the pooled results, we found that DM patients had a higher incidence of cardiac death, a higher incidence of stent thrombus, a higher incidence of TLR, a higher incidence of TVR, a higher incidence of MI, and a higher incidence of MACE. Based on the results, we believed that DM indeed affected the outcomes after PCI even with DES.,41 All these factors may impact the following revascularization. A greater plaque burden, higher propensity to plaque rupture, enhanced prothrombotic status, exuberant neointimal hyperplasia, more aggressive pattern of atherosclerosis, and endothelial dysfunction could be seen in the inflammatory environments in DM patients. and all these features would help to prompt the occurrence of restenosis after PCI. All results above would explain the reasons why DM patients experienced more adverse events.Autopsy and angiographic studies have demonstrated that DM patients were more likely to have higher rates of left main stenosis, chronic total occlusions, diffuse and multivessel disease, smaller vessel size, and longer lesion length.4.2 showed that insulin therapy was not statistically associated with increased propensity for stent thrombus, although ITDM remained at higher risk for other adverse cardiovascular events, which was similar to our study. Baseline characteristics were similar between ITDM and NITDM patients except more ITDM patients experienced renal insufficiency (11.8% vs. 2.3%), and this may explain the increased risk of stent thrombus and other adverse events.At 2-year follow-up, ITDM patients were more likely to experience stent thrombus, TVR, and MACE. Jain et al Insulin has both proatherogenic and antiatherogenic properties, which would differentially modify the risk of cardiovascular events, depending on the presence of insulin resistance and hyperinsulinemia.For DM patients, insulin resistance has been associated with detrimental biological processes such as impaired vascular production of nitric oxide and increased levels of endothelin-I and angiotensin-II.P = .263; stent thrombus, P = .995; MI, P = .548; TVR, P = .439; cardiac death, P = .263) except TLR  and MACE . The observation that the risk of serious cardiovascular events was similar between NITDM and non-DM patients might have some clinical implications in selecting the coronary revascularization strategy for patients with DM.In terms of the cumulative risk of adverse events, the risk of NITDM patients and non-DM patients was almost similar (cardiac death, We have several limitations in our study. First, the study design is retrospective. In retrospective study, selection bias would occur. But our baseline characteristics were similar between 2 groups. This may reduce the bias to some extent. Second, sample size was not large. Since the data were from a single center, unlike those from multiple-center collaboration, the limitations existed. However, the data reflected our own experience in Chinese population and indeed more future studies with large sample size and pooled results from multiple center were needed to confirm our results.5DM could be served as an independent predictor of adverse outcomes after drug-DES implantation. These patients should be reexamined more frequently and pay more attention to their stents.The authors thank Dr. Jonas Cook for language editing.


Awake surgery with electrocorticosubcortical stimulation is the golden standard treatment for gliomas in eloquent areas. Preoperatively, mostly mild cognitive disturbances are observed with postoperative deterioration. We describe pre- and postoperative profiles of 4 patients (P1–P4) with gliomas in “critical” language areas  undergoing awake surgery to get insight into the underlying mechanism of neuroplasticity. Neuropsychological examination was carried out preoperatively (at T1) and postoperatively . At T1, cognition of P1 was intact and remained stable. P2 had impairments in all cognitive domains at T1 with further deterioration at T2 and T3. At T1, P3 had impairments in memory and executive functions followed by stable recovery. P4 was intact at T1, followed by a decline in a language test at T2 and recovery at T3. Intraoperatively, in all patients language positive sites were identified. Patients with gliomas in “critical” language areas do not necessarily present cognitive disturbances. Surgery can either improve or deteriorate (existing) cognitive impairments. Several factors may underlie the plastic potential of the brain, for example, corticosubcortical networks and tumor histopathology. Our findings illustrate the complexity of the underlying mechanism of neural plasticity and provide further support for a “hodotopical” viewpoint. Awake surgery is considered the golden standard treatment for low-grade gliomas (LGG) in eloquent regions to optimize tumor resection while preserving neurological and cognitive functions and hence quality of life , 2. HoweEloquent regions typically include the left dominant perisylvian brain regions. DES has provided evidence for a “hodotopical”  view of the organization of brain functions as opposed to a “topological” viewpoint  –8. LanguThis is a follow-up study of 4 patients that we selected based on tumor localization in perisylvian language areas. The patients  were diagnosed with a glioma in the language dominant left hemisphere as identified with fMRI  with spontaneous speech monitoring during resection .http://www.osirix-viewer.com/). Postoperative MRI scans were assessed at 6 months after surgery. The extent of the resection was calculated as the fraction (%) of the difference between the preoperative and postoperative volume divided by the preoperative volume. The histological type of the tumor  and the World Health Organization (WHO) grade (2007) were determined by a neuropathologist, from tissue obtained during the tumor resection.Localization of the tumor was determined by a neuroradiologist using 3D T1-weighted images and 2D T2-weighted images. The pre- and postoperative tumor volume was calculated by manual delineation of 3-dimensional deviant signal intensity on T2-weighted MR images using Osirix version 4.1.2. ( Language tests are as follows: Boston Naming Test (BNT) or object naming (DuLIP), action naming, category and letter fluency, and Aachen Aphasia Test (AAT) subtests: repetition, writing to dictation, reading aloud, and Token Test. Memory tests are as follows: 15-word test ; digit span. Attentional and executive functions tests are as follows: design fluency, Trail Making Tests A and B, and Stroop Color Word Tests I–III. Based on the normative data, z-scores were computed to compare performance of the patients to healthy controls. A clinical impairment is reflected by a z-score between −1.5 and −2; a pathological impairment is reflected by a z-score of ≥−2. Postoperatively, P1 and P2 were tested at 6 weeks and 6 months and P3 and P4 at 3 months and 1 year. The study was approved by the Ethical Committee of Erasmus MC Rotterdam and University of Brussels. All patients gave written informed consent.Pre- and postoperatively, we administered an extensive neuropsychological test-protocol . Languag3 to 108 cm3. Pathological examination of tumor tissue obtained during resection revealed a LGG (WHO grade II) in P1 and P3 and a HGG in P2 (WHO grade IV) and in P4 (WHO grade III). The extent of resection (EoR) ranged from 58 to 89%. P1, P2, and P4 underwent postoperative radiotherapy (33 fraction doses of 1.8 Gy). All patients used anticonvulsants pre- and postoperatively (see Tumor volume ranged from 1.46 cmvely see .z ≥ −1.5). During operation, speech arrest occurred after stimulation at the precentral gyrus (primary motor cortex) at the level of the mouth. Postoperatively, at 6 weeks a clinical deficit in a memory test was observed  which recovered at 6 months. No other cognitive deficits were observed (z ≥ −1.5).Preoperatively, the cognitive functions of P1 were intact , and attention and executive deficits . During surgery cortical stimulation in the posterior superior temporal gyrus/angular gyrus triggered speech arrest. During stimulation of the AF, phonemic paraphasia occurred; these increased during resection near the AF, at which point resection was terminated. At 6 weeks postoperatively new deficits were found in language , in memory , and in attention and executive functions . There was an increase of the preoperative deficits in language  and in attention and executive functions . At 6 months postoperatively, improvement was observed in 1 subtest within the language domain (action naming: z = −1.37) and in the attention and executive functions (TMT A: z = −1.50), but further deterioration was found in another subtest (design fluency: productivity: z = −2.05).Preoperatively, P2 had clinically or pathologically significant impairments in language  and had selective pathological impairments in executive functioning . During surgery speech arrest occurred with stimulation of the inferior frontal gyrus, below the motor cortex, and the parietal lobe (see z = −0.50) and executive functioning , which remained stable during the follow-up of 1 year . No other impairments were present.Preoperatively P3 was clinically impaired in memory  which had recovered at 1 year (z = −1.39) and in attention and executive functions (TMT BA: z = −1.60) which also recovered at 1 year (z = −0.60).Preoperatively, P4 had no cognitive disorders. During surgery speech arrest was found when the inferior frontal gyrus was stimulated, below the motor cortex and in the temporal lobe. Phonemic paraphasia was elicited in the parietal lobe. Resection was terminated when perseverations occurred. At 3 months postoperatively, P4 developed a deficit in language  a language disorder was present preoperatively. Our study revealed mixed cognitive profiles at pre- and postoperative time-points. Two patients (P1 and P4) showed relatively intact cognitive performance, which may be explained by neural plasticity . By contrast, the other two cases (P2 and P3) demonstrated impairments in several cognitive domains pre- and/or postoperatively. These different findings are in line with a “dynamic” or “hodotopical” brain as opposed to a “static” or “topological” viewpoint . SeveralP1 with a tumor in Wernicke's area appeared to have intact cognition, apart from a temporary clinical memory deficit at 6 weeks postoperatively. Intact cognition after glioma surgery in Wernicke's area has been reported previously . By contApart from localization and the intraoperative procedure, the differential pre- and postoperative cognitive profiles in our patients could be attributed to tumor related factors, such as tumor grade. Noll et al.  found thAnother intervening factor influencing preoperative cognitive performance could be tumor volume. Habets et al.  found thEoR may have played a role as P2 and P4 underwent a more extensive tumor resection than P1 and P3. However, a recent follow-up study did not reveal a relation between EoR and cognitive decline . CurrentIn general, (stimulation-induced) seizures and the use of anticonvulsants can be a risk factor for deficits in cognitive performance . DeficitFinally, some other factors should be taken into account when interpreting our results. Handedness may have interfered with the results, as P1 was left-handed as opposed to P2– P4. Language organization in left-handed people is not always consistent and can be represented in a more widespread network than in right-handed people . HoweverPatients with brain tumors in “classical” language areas do not necessarily present language (or other cognitive) disturbances. Surgery can either improve or deteriorate (existing) cognitive (impairments) functions. The findings of these case studies provide therefore further support for neural plasticity within a “hodotopical” framework. It remains uncertain to which extent and which factors, such as localization, tumor grade, volume, EoR, and/or adjuvant therapy, contribute to neural plasticity. Hence, an extensive examination of cognitive functions with larger (sub)groups taking into account localization, tumor, and treatment related factors will elucidate prognostic factors of the plastic potential of the brain.


Host jump can result in deadly pandemic events when avian influenza A viruses broaden their host specificity and become able to infect mammals, including humans. Haemagglutinin—the major capsid protein in influenza A viruses—is subjected to high rate mutations, of which several occur at its “head”: the receptor-binding domain that mediates specific binding to host cell receptors. Such surface-changing mutations may lead to antigenically novel influenza A viruses hence in pandemics by host jump and in vaccine escape by antigenic drift. Changes in haemagglutinin surface electrostatics have been recently associated with antigenic drift and with clades evolution and spreading in H5N1 and H9N2 viruses. We performed a comparative analysis of haemagglutinin surface electrostatics to investigate clustering and eventual fingerprints among representative pandemic (H5 and H7) and nonpandemic (H4 and H6) avian influenza viral subtypes. We observed preferential sorting of viruses isolated from mammalian/human hosts among these electrostatic clusters of a subtype; however, sorting was not “100% specific” to the different clusters. Therefore, electrostatic fingerprints can help in understanding, but they cannot explain alone the host jumping mechanism. Influenza A viruses cause respiratory infections ranging from asymptomatic to deadly and can infect both birds and mammals, thus representing a dangerous threat to human health and poultry industry. Concerning humans, severe influenza A widespread outbreaks (pandemics) can result in the death of tens of million people worldwide, such as what occurred in 1918, 1957, and 1968 . Influenα2-6 SA, and thus improved affinity to the human host [So far up to 18 major subtypes (H1 to H18) have been classified and deposited in public databases such as the Influenza Research Database (IRD) ; among aman host –16. Receman host . Indeed,We recently reported that when comparing the HA receptor-binding domain (RBD), electrostatic closeness can group haemagglutinins from different AI virus phylogenetic groups. In particular, H5 (which belongs to HA Group 1) was found to be quite closer to H9 (HA Group 2) than to H2 (same group) . Deeper Indeed, electrostatics is an important player in the modulation of interactions at all subcellular levels. In particular, electrostatic interactions are ubiquitous in proteins and dictate stability and function. As reported by Ritchie and Webb  structurElectrostatics is able to keep influenza virus Matrix Protein M1 conformation stable at different pH values , and divHowever, and in spite of being so important for protein function and dysfunction, electrostatics is only one player among other  modulators of the protein surface features (hence of protein interactions). Indeed, solvent accessible surface area and the balance between hydrophilic and hydrophobic patches and side chain orientation at specific linear or conformational epitopes may play a pivotal role as well. Therefore, having found and confirmed relationship with virus clades evolution , 19, we In this work, we studied electrostatic variation among four HA subtypes, of which two (H5 and H7) representing a model for “pandemic” subtypes and two H4 and H6) for “non/poorly pandemic” ones. We started by comparative analysis of HA1 chains, as HA1 contains the RBD, where electrostatic fingerprints were found , 19, whi and H6 fhttps://www.fludb.org. Target protein sequences were modeled on best available structure templates using SWISS-MODEL [Data were obtained from the NIAID Influenza Research Database (IRD)  through SS-MODEL . Model qSS-MODEL .kBT/e. PDB2PQR [εp = 2 and εs = 78.5 were chosen for, respectively, the protein and the solvent [T = 298.15 K. Probe radius for dielectric surface and ion accessibility surface were set as r = 1.4 Å and r = 2 Å, respectively. Electrostatic distance (ED):Isopotential contours were calculated using UCSF Chimera v. 1.11.2  via Adap PDB2PQR  was used PDB2PQR . Interio solvent , 38, 39,A server , which cIn two recent studies, electrostatic distance (ED) analysis of the different AI virus clades and subclades was used to integrate phylogenetic analyses, and comparative analysis of electrostatic isocontours unveiled “charge redistribution” events at the RBD surface that suggested a molecular rationale for evolutionary drift underlying adaptation and spreading of successful clades in H5N1 and H9N2 , 19. ThiWe populated a sixty-four dataset for ED analysis  by overrepresenting (thirty-five) h/m-host subpopulation with respect to a-host viruses (twenty-nine). In early analyses, the HA1 mature chain for each haemagglutinin sequence was considered. Since the input for the ED analysis consists of pdb structures rather than sequences, a structural model for each HA1 chain was obtained by homology modeling with high confidence, because of the very high identity with the available structural template. This notwithstanding, model quality was assessed via QMEAN  prior toThe electrostatic isocontour for each HA1 structural model was obtained , and four 90°-stepwise views are presented as a full-view flat picture for each isocontour. A multipage figure, depicting the complete analysis with all sixty-four viruses, is presented as Supplementary The ED analysis performed with H5 was repeated with a same-size dataset of viruses from another AI subtype having a well-known track of infections in humans and other mammals, that is, H7 (see introduction). Several different electrostatic groups are found also with H7; in particular, the three largest groups (G1–G3) reveal a relatively low intragroup ED , more than 43000 published papers can be retrieved. Papers are still thousands with additional search keywords such as “pandemic” or “evolution” or hundreds when adding, for example, “antigenic drift”, “immune escape”, or “host specificity”. The most of such works were based on wet biology analyses and/or sequence based phylogenetic trees; however, in spite of intriguing evidence having emerged from multiple, very good works, no paper could place any final explanation, or even to clearly identify the major player for viral clade evolution and spreading or for host specificity shift. Considering that structure based, more systematic analysis was needed, we recently focused on comparative electrostatic analysis of the haemagglutinin surface. This allowed us to demonstrate that, in H5N1, electrostatic fingerprints and a progressive change in charge distribution are clearly related to clades and subclades evolution and spreading in different AI types , more th. In this


Similarity transformations are used to transform the governing nonlinear boundary layer equations for momentum, thermal energy and concentration to a system of nonlinear ordinary coupled differential equations with fitting boundary conditions. The transmuted model is shown to be controlled by a number of thermo-physical parameters, viz. the magnetic parameter, thermal convective parameter, mass convective parameter, radiation-conduction parameter, heat generation parameter, Prandtl number, Lewis number, Brownian motion parameter, thermophoresis parameter, chemical reaction parameter and pressure gradient parameter. Numerical elucidations are obtained with the legendary Nactsheim-Swigert shooting technique together with Runge–Kutta six order iteration schemes. Comparisons with previously published work are accomplished and proven an excellent agreement.The present study analyzed numerically magneto-hydrodynamics (MHD) laminar boundary layer flow past a wedge with the influence of thermal radiation, heat generation and chemical reaction. This model used for the momentum, temperature and concentration fields. The principal governing equations is based on the velocity  Koh and Hartnett uw(x) >0B =  is applied in the y-direction.Here in equation  the 3rd Again in equation  the 2nd Also in equation  the 2nd And the boundary condition for the model is;In order to conquers a similarity solution to eqs.  to 4) w w4) withue(x) = axm, uw(x) = cxm, where a, c and m (0 ≤ m ≤ 1) are positive constant. Therefore, the constant moving parameter λ is defined as λ = c/a, whereas λ < 0 relates to a stretching wedge, λ > 0 relates to a contracting wedge, and λ = 0 corresponds to a fixed wedge, respectively.For the similarity solution of equation  to 4) w w4) withFrom the above transformations the non-dimensional, nonlinear, coupled ordinary differential equations are obtained as;η and the parameters are defined as:The transformed boundary conditions are as follows;Magnetic parameter, Pressure gradient parameter, Grashof number, Modified Grashof number, Thermal convective parameter, Mass convective parameter, Local Reynolds number, Prandtl number, Heat source parameter, Lewis number, Brownian motion parameter, Thermophoresis parameter, Radiation parameter, Chemical reaction parameter, et al. [M is a local magnetic body force number  and λTnd Gorla ).η. A method for the numerical solution of the differential equations of the boundary layer type was presented by Philip R. Nachtsheim and Paul Swigert in 1965. For the purpose of present problem, we applied the Nacthsheim-Swigert iteration technique.The non dimensional, nonlinear, coupled ordinary differential equations  to 9) w w9) withIn shooting method, the missing (unspecified) initial condition at the initial point of the interval is assumed and the differential equation is integrated numerically as an initial value problem to the terminal point. The accuracy of the assumed missing initial condition is then checked by comparing the calculated value of the dependent variable at the terminal point with its given value there. If a difference exists, another value of the missing initial condition must be assumed and the process is repeated. This process is continued until the agreement between the calculated and the given condition at the terminal point is within the specified degree of accuracy. For this type of iterative approach, one naturally inquires whether or not there is a systematic way of finding each succeeding (assumed) value of the missing initial condition.The boundary conditions equation  associatf//(0), θ/(0) and φ/(0).However, this is not generally the case. Hence a method must be devised to logically estimate the new surface boundary conditions for the next trial integration. Asymptotic boundary value problems such as those governing the boundary layer equations are further complicated by the fact that the outer boundary condition is specified at infinity. In the trial integration infinity is numerically approximated by some large value of the independent variable. There is no a priori general method of estimating this value. Selection of too small a maximum value for the independent variable may not allow the solution to asymptotically converge to the required accuracy. Selecting a large value may result in divergence of the trial integration or in slow convergence of surface boundary conditions required satisfying the asymptotic outer boundary condition. Selecting too large a value of the independent variable is expensive in terms of computer time. Nachtsheim-Swigert developed an iteration method, which overcomes these difficulties. Extension of the iteration shell to above equation system of differential equations  is straif/), temperature (θ) and concentration (φ) have been computed for resultant principal parameters as the Magnetic parameter M, pressure gradient parameter β, Thermal convective parameter λT, Mass convective parameter λM, local Reynolds number Re, Prandtl number Pr, Heat source parameter Q, Lewis number Le, Brownian motion parameter Nb, thermophoresis parameter Nt, radiation parameter R and chemical reaction parameter γ respectively. To assess the accuracy of the numerical results the Skin friction coefficient f//(0) compared with previous literatures. And excellent agreement is observed.In order to investigate the physical representation of the problem, the numerical values of velocity (f//(0) for several values of when λ = λT = λM = M = R = Q = γ = 0 have been compared White [The prediction of Skin friction coefficient ed White , Yih [30ed White , and Khaed White  and showλT). The momentum boundary layer follows the boundary condition due to the shrinking wedge. An increase in thermal convective  parameter is observed to strongly accelerate the flow. Therefore the velocity profiles are rises within the boundary layer as the λT increase. Figure λM). Near the shirking wedge surface it was found that, with increasing the mass convective (species buoyancy) parameter serves to increase the velocity profile. Therefore increasing species buoyancy force therefore only aids momentum and increases velocity boundary layer thickness further from the sheet. It was also observed that the momentum boundary layer follows the boundary condition due to the shrinking wedge. Figure Q). Then for above case it is observed that velocity profiles are increases as the Q increase. It was also found that the shrinking wedge follows the boundary condition.Figure R). An increase in radiation parameter (R) is found to slightly upturn velocity values close to the sheet because of the flow is accelerated therefore momentum boundary layer thickness is therefore increased strongly after some distance from the wall. Also the shrinking wedge follows the boundary condition appropriately. With increasing thermal convective parameter as plotted in Figure λM. An increase in mass convective (species buoyancy) parameter, as shown in Figure Figure Q)  in Figure Figure γ. Nano-particle concentration and concentration boundary layer thickness is decreased with strong chemical reaction closer to the sheet surface, whereas the opposite trend is computed further from the sheet surface. Therefore finally concentration boundary layer thickness is increases. Figure R. Through rising radiation (R), concentration in the nanofluid is significantly diminished.Figure In this study, the governing equations for the considered MHD radiative, heat-generating and chemical reacting nanofluid flow past a wedge is presented. A numerical representation has been developed for the boundary layer flow past a wedge with the influence of thermal radiation, heat generation and chemical reaction. It was found a good accuracy of skin friction coefficient which is compared with the previous studies ,29-31.λT, λM, Q and R increase correspondingly.The velocity profiles are rises within the boundary layer as the With increasing independently thermal convective parameter and radiation than temperature increased significantly. Whereas the opposite effect has been found for mass convective (species buoyancy) and heat generation parameter respectively.A strong decrease in the nano-particle concentration is caused by increasing the thermal convective parameter and radiation individually. But it rises for increasing mass convective, chemical reaction and heat generation parameter.The significant findings of present study are given below:A, positive constantBo, magnetic inductionC, positive constantC, nanoparticle concentrationCw, nanoparticle concentration at wedgeC∞, ambient nanoparticle concentration as y tends to infinitycp, specific heat capacityDB, Brownian diffusion coefficientDT, thermophoresis diffusion coefficientG, acceleration due to gravityGr, Grashof numberGm, Modified Grashof numberKr, rate of chemical reactionLe, Lewis numberM, positive constantM, magnetic parameterNb, Brownian motion parameterNt, Thermophoresis parameterP, fluid pressurePr, Prandtl numberQ, heat source parameterQo, heat generation constantRe, local Reynolds numberT, fluid temperatureTw, temperature at the wedge surfaceT∞, ambient temperature as y tends to infinityx, y, Cartesian coordinates measured along wedgeu, v, velocity components along x and y axes respectivelyGreek symbolsΝ, dynamic and kinematic viscosities(ρc)p, effective heat capacity of the nanofluid(ρc)f, heat capacity of the fluidσ, charge densityα, thermal diffusivityβ, pressure gradient parameterβT, co-efficient of thermal expansionβC, co-efficient of mass expansionλ, constant moving parameterγ, Chemical reaction parameterλT, thermal convective parameterλM, mass convective parameterη, similarity variableψ, stream functionf(η), dimensionless velocityθ(η), dimensionless temperatureφ(η), dimensionless concentration