Genomic innovations, transcriptional plasticity and gene loss underlying the evolution and divergence of two highly polyphagous and invasive Helicoverpa pest species.

BACKGROUND: Helicoverpa armigera and Helicoverpa zea are major caterpillar pests of Old and New World agriculture, respectively. Both, particularly H. armigera, are extremely polyphagous, and H. armigera has developed resistance to many insecticides. Here we use comparative genomics, transcriptomics and resequencing to elucidate the genetic basis for their properties as pests. RESULTS: We find that, prior to their divergence about 1.5 Mya, the H. armigera/H. zea lineage had accumulated up to more than 100 more members of specific detoxification and digestion gene families and more than 100 extra gustatory receptor genes, compared to other lepidopterans with narrower host ranges. The two genomes remain very similar in gene content and order, but H. armigera is more polymorphic overall, and H. zea has lost several detoxification genes, as well as about 50 gustatory receptor genes. It also lacks certain genes and alleles conferring insecticide resistance found in H. armigera. Non-synonymous sites in the expanded gene families above are rapidly diverging, both between paralogues and between orthologues in the two species. Whole genome transcriptomic analyses of H. armigera larvae show widely divergent responses to different host plants, including responses among many of the duplicated detoxification and digestion genes. CONCLUSIONS: The extreme polyphagy of the two heliothines is associated with extensive amplification and neofunctionalisation of genes involved in host finding and use, coupled with versatile transcriptional responses on different hosts. H. armigera's invasion of the Americas in recent years means that hybridisation could generate populations that are both locally adapted and insecticide resistant.

Chemical vapor sensing with monolayer MoS2.

Two-dimensional materials such as graphene show great potential for future nanoscale electronic devices. The high surface-to-volume ratio is a natural asset for applications such as chemical sensing, where perturbations to the surface resulting in charge redistribution are readily manifested in the transport characteristics. Here we show that single monolayer MoS(2) functions effectively as a chemical sensor, exhibiting highly selective reactivity to a range of analytes and providing sensitive transduction of transient surface physisorption events to the conductance of the monolayer channel. We find strong response upon exposure to triethylamine, a decomposition product of the V-series nerve gas agents. We discuss these results in the context of analyte/sensor interaction in which the analyte serves as either an electron donor or acceptor, producing a temporary charge perturbation of the sensor material. We find highly selective response to electron donors and little response to electron acceptors, consistent with the weak n-type character of our MoS(2). The MoS(2) sensor exhibits a much higher selectivity than carbon nanotube-based sensors.

Bilayer graphene grown on 4H-SiC (0001) step-free mesas.

We demonstrate the first successful growth of large-area (200 × 200 µm(2)) bilayer, Bernal stacked, epitaxial graphene (EG) on atomically flat, 4H-SiC (0001) step-free mesas (SFMs) . The use of SFMs for the growth of graphene resulted in the complete elimination of surface step-bunching typically found after EG growth on conventional nominally on-axis SiC (0001) substrates. As a result heights of EG surface features are reduced by at least a factor of 50 from the heights found on conventional substrates. Evaluation of the EG across the SFM using the Raman 2D mode indicates Bernal stacking with low and uniform compressive lattice strain of only 0.05%. The uniformity of this strain is significantly improved, which is about 13-fold decrease of strain found for EG grown on conventional nominally on-axis substrates. The magnitude of the strain approaches values for stress-free exfoliated graphene flakes. Hall transport measurements on large area bilayer samples taken as a function of temperature from 4.3 to 300 K revealed an n-type carrier mobility that increased from 1170 to 1730 cm(2) V(-1) s(-1), and a corresponding sheet carrier density that decreased from 5.0 × 10(12) cm(-2) to 3.26 × 10(12) cm(-2). The transport is believed to occur predominantly through the top EG layer with the bottom layer screening the top layer from the substrate. These results demonstrate that EG synthesized on large area, perfectly flat on-axis mesa surfaces can be used to produce Bernal-stacked bilayer EG having excellent uniformity and reduced strain and provides the perfect opportunity for significant advancement of epitaxial graphene electronics technology.

NK cell transcripts and NK cells in kidney biopsies from patients with donor-specific antibodies: evidence for NK cell involvement in antibody-mediated rejection.

To explore the mechanisms of antibody-mediated rejection (ABMR) in kidney transplants, we studied the transcripts expressed in clinically indicated biopsies from patients with donor-specific antibody (DSA). Comparison of biopsies from DSA-positive versus DSA-negative patients revealed 132 differentially expressed transcripts: all were associated with class II DSA but none with class I DSA. Many transcripts were expressed in DSA-positive ABMR but were also expressed in T-cell-mediated rejection (TCMR), reflecting shared molecular features. Removal of shared transcripts created 23 DSA selective transcripts (DSASTs). Some DSASTs (6/23) showed selective high expression in NK cells, whereas others (8/23) were expressed in endothelium or in endothelium plus other cell types (7/23). Of 145 biopsies ranked by DSAST expression, the 25 with highest DSAST expression primarily consisted of ABMR (22/25, 88%), either C4d-positive or C4d-negative. By immunostaining, CD56+ and CD68+ cells in peritubular capillaries, but not CD3+ cells, were increased in ABMR compared to TCMR, compatible with a role for NK cells, as well as macrophages, as effectors in endothelial injury during ABMR. Thus, the strategy of using DSASTs in the biopsy to identify mechanism-related transcripts in biopsies from patients with clinical phenotypes indicates the selective involvement of NK cells in ABMR.

Absence of donor-specific anti-HLA antibodies after ABO-incompatible heart transplantation in infancy: altered immunity or age?

Specific B-cell tolerance toward donor blood group antigens develops in infants after ABO-incompatible heart transplantation, whereas their immune response toward protein antigens such as HLA has not been investigated. We assessed de novo HLA-antibodies in 122 patients after pediatric thoracic transplantation (28 ABO-incompatible) and 36 controls. Median age at transplantation was 1.7 years (1 day to 17.8 year) and samples were collected at median 3.48 years after transplantation. Antibodies were detected against HLA-class I in 21 patients (17.2%), class II in 18 (14.8%) and against both classes in 10 (8.2%). Using single-antigen beads, donor-specific antibodies (DSAs) were identified in six patients (all class II, one additional class I). Patients with DSAs were significantly older at time of transplantation. In patients who had undergone pretransplant cardiac surgeries, class II antibodies were more frequent, although use of homografts or mechanical heart support had no influence. DSAs were absent in ABO-incompatible recipients and class II antibodies were significantly less frequent than in children with ABO-compatible transplants. This difference was present also when comparing only children transplanted below 2 years of age. Therefore, tolerance toward the donor blood group appears to be associated with an altered response to HLA beyond age-related effects.

Effects of increased surgical trauma on rates of tooth movement and apical root resorption in foxhound dogs.

PURPOSE: To experimentally determine the effects of increased surgical trauma on the rates of tooth movement and apical root resorption. Two surgical techniques for rapid protraction of multi-rooted teeth in foxhound dogs immediately following premolar extraction were compared. METHODS: Split-mouth design to randomly assign two surgical techniques [periodontal ligament distraction (RAP side) and a modified form of dentoalveolar distraction (RAP+ side)] to the maxillary quadrants. First premolars were extracted, and second premolars were protracted 0.5 mm per day for 15 days using a custom made jack-screw distractor. Serial caliper and radiographic measurements were performed to quantify tooth movements and apical root resorption. RESULTS: Both techniques demonstrated significant movement of the crown and apex. The second premolar crowns were protracted significantly more on the RAP+ side (2.9 mm) than on the RAP (1.8 mm) side. The premolars on both sides demonstrated significant tipping (4.3 and 3.9 degrees for the RAP+ and RAP sides, respectively). The distal root apex showed almost twice as much apical root resorption than the mesial root apex, but resorption was limited (<0.16 mm) and not statistically different between sides. CONCLUSIONS: Increased surgical trauma increased the rate and, ultimately, the amount of tooth movement. The heavy forces used to protract the teeth produced statistically, but not clinically, significant apical root resorption on the mesial and distal roots of the maxillary second premolars.

Antibody-mediated microcirculation injury is the major cause of late kidney transplant failure.

We studied the phenotype of late kidney graft failure in a prospective study of unselected kidney transplant biopsies taken for clinical indications. We analyzed histopathology, HLA antibodies and death-censored graft survival in 234 consecutive biopsies from 173 patients, taken 6 days to 31 years posttransplant. Patients with late biopsies (>1 year) frequently displayed donor-specific HLA antibody (particularly class II) and microcirculation changes, including glomerulitis, glomerulopathy, capillaritis, capillary multilayering and C4d staining. Grafts biopsied early rarely failed (1/68), whereas grafts biopsied late often progressed to failure (27/105) within 3 years. T-cell-mediated rejection and its lesions were not associated with an increased risk of failure after biopsy. In multivariable analysis, graft failure correlated with microcirculation inflammation and scarring, but C4d staining was not significant. When microcirculation changes and HLA antibody were used to define antibody-mediated rejection, 17/27 (63%) of late kidney failures after biopsy were attributable to antibody-mediated rejection, but many were C4d negative and missed by current diagnostic criteria. Glomerulonephritis accounted for 6/27 late losses, whereas T-cell-mediated rejection, drug toxicity and unexplained scarring were uncommon. The major cause of late kidney transplant failure is antibody-mediated microcirculation injury, but detection of this phenotype requires new diagnostic criteria.

De novo donor-specific antibody at the time of kidney transplant biopsy associates with microvascular pathology and late graft failure.

We studied whether de novo donor-specific antibodies (DSA) in sera from patients undergoing kidney transplant biopsies associate with specific histologic lesions in the biopsy and prognosis. DSA were assessed in 145 patients at the time of biopsy between 7 days to 31 years posttransplant. DSA was detected in 54 patients (37%), of which 32 represented de novo DSA. De novo DSA was more frequent in patients having late biopsies (34%) versus early biopsies (4%), and was usually either against class II alone or class I and II but rarely against class I alone. Microcirculation inflammation (glomerulitis, capillaritis) and damage (glomuerulopathy, capillary basement membrane multilayering), and C4d staining were associated with de novo DSA. However, the degree of scarring, arterial fibrosis and tubulo-interstitial inflammation did not correlate with the presence of de novo DSA. De novo DSA correlated with reduced graft survival after the biopsy. Thus, de novo DSA at the time of a late biopsy for clinical indication is primarily against class II, and associates with microcirculation changes in the biopsy and subsequent graft failure. We propose careful assessment of de novo DSA, particularly against class II, be performed in all late kidney transplant biopsies.

Cost-benefit analysis for a lead wheel weight phase-out in Canada.

Lead wheel weights (LWWs) have been banned in Europe, and some US States, but they continue to dominate the market in Canada. Exposure to lead is associated with numerous health impacts and can result in multiple and irreversible health problems which include cognitive impairment when exposure occurs during early development. Such impacts incur high individual and social costs. The purpose of this study was to assess the costs and public health benefits of a Risk Management Strategy (RMS) that would result from a LWW phase-out in Canada and compare this to a Business-As-Usual (BAU) scenario. The contribution of LWWs to lead concentrations in media including roadway soil/dust, ambient and indoor air, and indoor dust were estimated. The Integrated Exposure Uptake Biokinetic Model for Lead in Children (IEUBK) was used to develop estimates for the blood lead levels (BLLs) in children (µg/dL) associated with the BAU and the RMS. The BLLs estimated via the IEUBK model were then used to assess the IQ decrements associated with the BAU that would be avoided under the RMS. The subsequent overall societal benefits in terms of increased lifetime earning potential and reduced crime rate, were then estimated and compared to industry and government costs. LWWs form 72% of the Canadian wheel weight market and >1500 tonnes of lead as new LWWs attached to vehicles enters Canadian society annually. We estimate that 110-131 tonnes of lead in detached WWs are abraded on roadways in Canada each year. A LWW phase-out was predicted to result in a drop in pre-school BLLs of up to 0.4 µg/dL. The estimated net benefits associated with the RMS based on cognitive decrements avoided and hence increased lifetime earning potential (increased productivity) and reduced crime are expected to be: C$248 million (8% discount rate) to C$1.2 billion (3% discount rate) per year.

Identification of a calcium-inducible, cyclosporine sensitive element in the IFN-gamma promoter that is a potential NFAT binding site.

Cyclosporine (CsA) inhibits cytokine transcription by preventing the activation of key promoter sites, in particular the binding of nuclear factor of activated T cells (NFAT) to the IL-2 NFAT site and the "P" site in IL-4. To identify potential NFAT-like sites in the IFN-gamma promoter, we sought areas of homology with the known sites in other promoters. In the promoter region of the mouse and human IFN-gamma gene, we identified two repeats of a consensus sequence ATTTCCnnT, designated P1 and P2 because of their homology to the calcium-inducible and CsA-sensitive "P" sequences in the IL-4 promoter. In electrophoretic mobility shift assay (EMSA), a probe containing the second P sequence "P2" in the human IFN-gamma gene bound nuclear proteins from stimulated, but not unstimulated, humans T cells. The cytosol of unstimulated cells contained similar binding activity that decreased after stimulation, indicating that this binding activity translocated to the nucleus after stimulation. CsA inhibited nuclear translocation. Competition studies demonstrated that oligomers containing the sequences P1 and P2 in IFN-gamma gene, the NFAT site in the IL-2 gene, and the IL-4 P site competed with the P2 probe for protein binding, whereas an oligomer containing mutations in the P2 site did not. Addition of anti-NFAT antiserum altered protein binding to P2, indicating that the proteins were either identical or related to NFAT. Stimulation of T cells transfected with constructs containing three copies of the P2 sequence enhanced CAT activity in response to ionomycin, and this effect was blocked by CsA. These results suggest that the P2 sequence, and probably the P1 sequence, in the IFN-gamma promoter are NFAT binding sites and contribute to the calcium inducibility and CsA sensitivity of IFN-gamma production.

Synaptic numbers across cortical laminae and cognitive performance of the rat during ageing.

In this study, we have investigated the changes in the number of individual presynaptic boutons in the neocortex of rats and correlated them with cognitive performance. Brown Norway x Fischer 344 F1 hybrid rats, aged from one to 24 months, were used. Using synaptophysin as a marker for presynaptic boutons, we found that in the parietal II region of the neocortex an age-related decrease in the density of immunostained punctae representing presynaptic boutons occurred. Regression analysis showed that this decline in the number of presynaptic boutons correlates with ageing (r=0.495, P<0.05). Interestingly, we found that this age-related depletion of presynaptic boutons was more intense in the deeper cortical lamina, such as laminae V and VI (mean decrease of 18%), than in the superficial laminae (mean decrease of 8% in laminae I-IV). Using the Morris water maze test, we observed that young rats acquired the task at twice the speed of aged animals (48.9 +/- 9.0 s and 91.0 +/- 4.9 s for young and aged animals, respectively). Furthermore, at the end of the training period, the aged cohort still showed significantly higher escape latencies in the Morris water maze. The present findings support the concept that the decline in cognitive performances in ageing is related to the loss of synapses in the cerebral cortex.

Ecological risk assessment of endocrine disruptors.

The European Centre for Ecotoxicology and Toxicology of Chemicals proposes a tiered approach for the ecological risk assessment of endocrine disruptors, integrating exposure and hazard (effects) characterization. Exposure assessment for endocrine disruptors should direct specific tests for wildlife species, placing hazard data into a risk assessment context. Supplementing the suite of mammalian screens now under Organization for Economic Cooperation and Development (OECD) validation, high priority should be given to developing a fish screening assay for detecting endocrine activity in oviparous species. Taking into account both exposure characterization and alerts from endocrine screening, higher tier tests are also a priority for defining adverse effects. We propose that in vivo mammalian and fish assays provide a comprehensive screening battery for diverse hormonal functions (including androgen, estrogen, and thyroid hormone), whereas Amphibia should be considered at higher tiers if there are exposure concerns. Higher tier endocrine-disruptor testing should include fish development and fish reproduction tests, whereas a full life-cycle test could be subsequently used to refine aquatic risk assessments when necessary. For avian risk assessment, the new OECD Japanese quail reproduction test guideline provides a valuable basis for developing a test to detecting endocrine-mediated reproductive effects; this species could be used, where necessary, for an avian life-cycle test. For aquatic and terrestrial invertebrates, data from existing developmental and reproductive tests remain of high value for ecological risk assessment. High priority should be given to research into comparative endocrine physiology of invertebrates to support data extrapolation to this diverse fauna.

Purification and kinetic characterisation of juvenile hormone esterase from Drosophila melanogaster.

Juvenile hormone esterase (JHE) from the prepupal stage of Drosophila melanogaster was purified about 429-fold to near homogeneity by selective precipitations, isoelectric focussing, anion exchange and gel filtration chromatography. The KM and Vmax of the purified enzyme for juvenile hormone III (JHIII) hydrolysis are 89 nM and at least 590 nmol/min/mg, respectively. JHE also hydrolyses the artificial substrate alpha-naphthyl acetate with a KM of 120 micro M and a Vmax of at least 70 mumol/min/mg. Competition of JHIII hydrolysis by five juvenile hormones and twenty-four JH analogues showed JHE is highly selective for JHIII and JHIII bisepoxide (JHP3), and both may be in vivo substrates. Binding in the active site of JHE is promoted by structural features found in JHIII and JHB3 including the epoxide groups in their natural orientations, methyl (rather than ethyl) side-chains, and the 2E, 3 double bond that is conjugated with the ester group. Binding is reduced by almost any departure from these structural features of JH. Co-incubation of the haemolymph JH binding protein, lipophorin, with JHE indicates lipophorin might modulate JH hydrolysis by competition for binding of JH.

cDNA cloning, baculovirus-expression and kinetic properties of the esterase, E3, involved in organophosphorus resistance in Lucilia cuprina.

Resistance to organophosphorus insecticides (OPs) in the sheep blowfly, Lucilia cuprina, is associated with a non-staining phenotype of the carboxylesterase isozyme, E3 (E.C. 3.1.1.1). Here, we show that a member of alpha-esterase multigene family, Lc alpha E7, encodes E3. An Lc alpha E7 cDNA has been isolated from an OP-susceptible strain and expressed in a baculovirus. The expressed product is the same as E3 in its electrophoretic mobility and preference for alpha-over beta-naphthyl acetate as substrate. Its preference (kcat/K(m)) for a range of carboxylester substrates is alpha-naphthyl butyrate > alpha-naphthyl propionate > alpha-naphthyl acetate > methylthiobutyrate > p-nitrophenyl acetate. The enzyme is potently inhibited by OPs (ki [paraoxon] = 6.3 +/- 1.4 x 10(7)/M/min, ki [chlorfenvinphos] = 5.9 +/- 0.6 x 10(7)/M/min) and exhibits a high turnover of methylthiobutyrate (1009/s), consistent with its proposed homology to the ali-esterase that is thought to mutate to confer OP resistance in Musca domestica. E3 shares 64% amino acid identity with its Drosophila melanogaster homologue, Dm alpha E7, and is also closely related to other esterases involved in OP resistance such as the B1 esterase of Culex pipiens (38%) and E4 of Myzus persicae (30%).

Identification of a juvenile hormone esterase gene by matching its peptide mass fingerprint with a sequence from the Drosophila genome project.

Juvenile hormone esterase (JHE, EC 3.1.1.1) from whole Drosophila melanogaster prepupae has previously been purified by selective precipitations, isoelectric focussing and two column chromatography steps. JHE bands from dried silver-stained SDS-PAGE gels of that material were digested with trypsin. The masses of the tryptic digest peptides were determined by MALDI-TOF mass spectrometry. Only one predicted gene product (CG8425) from the D. melanogaster genome matches the JHE tryptic fingerprint with high confidence. This predicted JHE sequence includes features that are conserved among all active members of the serine carboxylesterase multigene family as well as features peculiar to JHEs from other species. Also we show that this JHE can be purified by an alternative method using anion exchange chromotography followed by trifluoromethylketone affinity chromatography. A cDNA encoding this JHE was isolated using 3' and 5' RACE. This sequence is in agreement with the Drosophila genome project's prediction except that the sixth predicted intron is not removed; instead there is a stop codon followed by a polyadenylation signal and a polyA tail.

Nifedipine inhibits responses to alpha-adrenoceptor stimulation in canine blood vessels: impact of heart failure.

The effects of nifedipine (10(-8) and 10(-7) M) on alpha-adrenergic responses of the dorsal pedal artery and saphenous vein were examined from dogs with pacing-induced heart failure. Two groups of dogs had their right ventricles paced at 250 beats/min: group (1) 1 week of pacing (mild heart failure) and group (2) paced for a mean period of 25.8 days (peak heart failure). Nifedipine non-competitively antagonised 6-allyl-2-amino-5,6,7,8-tetrahydro-4H- thiazolo[4,5-d]azepin dihydrochloride (BHT 920)-induced contractions to the same extent (i.e. at control, mild heart failure and peak heart failure) and IC50 values were as follows: for dorsal pedal artery 3.9 (1.8-6.1) nM, 4.4 (1.2-8.4) nM and 8.5 (2.9-38.9) nM, respectively; for saphenous vein 13.0 (4.6-26.0) nM, 13.0 (7.3-18.6) nM and 19.0 (9.3-32.8) nM, respectively). Before the onset of pacing, nifedipine did not affect concentration-effect curves generated to noradrenaline or phenylephrine in either the artery or the vein. After 1 week of pacing, nifedipine (10(-7) M) inhibited contractions to noradrenaline in the artery and the vein (70 +/- 5% for the artery and 51 +/- 4% for the vein). Nifedipine had no effect on phenylephrine-induced contractions. At peak heart failure, nifedipine inhibited both noradrenaline and phenylephrine contractions. These results indicate that nifedipine is useful in differentiating contractile activity of vascular smooth muscle with respect to alpha-adrenoceptor agonism.

Light scattering and gloss of an experimental quartz-filled composite.

For samples of polymethylmethacrylate with and without quartz filler, the inverse of the contrast-gloss ratio is shown to be related to surface roughness and to the optical scattering coefficient. This finding adds to the importance of optical scattering, which has been widely studied because of its relation to color and translucency of materials. Furthermore, optical scattering by composite fillers is shown to be linearly related to the concentration of the filler material within the range of concentrations studied. Quartz fillers were incorporated at concentrations from 5 to 20 weight percent and were short fibers or granular powder, with the granular particles ranging in median equivalent spherical diameter from 15 to 3.3 micron. The efficiency of optical scattering for the granular quartz filler increased as the size of the filler decreased.

Validation of self-reported transfusion histories in renal dialysis patients.

The purpose of this analysis was to assess the validity of self-reported transfusion histories in dialysis patients. Using data from a cross-sectional study of a dialysis population being investigated for hepatitis C virus (HCV) infection, the correspondence between self-reported transfusion history and transfusion records was explored. Demographic data and dialysis histories were examined in relation to the accuracy of self-reports. Overall, the questionnaire data and the blood bank records agreed for 89% of participants. The Kappa statistic was 0.72 (95% CI: 0.61, 0.83) indicating an acceptable level of agreement. The effect of non-differential exposure misclassification on the risk estimates for transfusion history as a determinant of HCV infection is demonstrated. Exploring the discrepancies between self-reports and documented transfusion histories underlines the need to communicate clearly medical interventions in chronically ill patients. Additionally, it suggests that studies into transfusion-acquired blood-borne pathogens should use all available information sources to establish exposure.