Impact of pediatric cardiac surgery regionalization on health care utilization and mortality.

OBJECTIVE: Regionalization directs patients to high-volume hospitals for specialized care. We investigated regionalization trends and outcomes in pediatric cardiac surgery. DATA SOURCES/STUDY SETTING: Statewide inpatient data from eleven states between 2000 and 2012. STUDY DESIGN: Mortality, length of stay (LOS), and cost were assessed using multivariable hierarchical regression with state and year fixed effects. Primary predictor was hospital case-volume, categorized into low-, medium-, and high-volume tertiles. DATA COLLECTION/EXTRACTION METHODS: We used Risk Adjustment for Congenital Heart Surgery-1 (RACHS-1) to select pediatric cardiac surgery discharges. PRINCIPAL FINDINGS: In total, 2841 (8.5 percent), 8348 (25.1 percent), and 22 099 (66.4 percent) patients underwent heart surgeries in low-, medium-, and high-volume hospitals. Mortality decreased over time, but remained higher in low- and medium-volume hospitals. High-volume hospitals had lower odds of mortality and cost than low-volume hospitals (odds ratio [OR] 0.59, P < 0.01, and relative risk [RR] 0.91, P < 0.01, respectively). LOS was longer for high- and medium-volume hospitals, compared to low-volume hospitals (high-volume: RR 1.18, P < 0.01; medium-volume: RR 1.05, P < 0.01). CONCLUSIONS: Regionalization reduced mortality and cost, indicating fewer complications, but paradoxically increased LOS. Further research is needed to explore the full impact on health care utilization.

Repellent and Attractive Effects of α-, ß-, and Dihydro-ß- Ionone to Generalist and Specialist Herbivores.

In plants, the oxidative cleavage of carotenoid substrates produces volatile apocarotenoids, including α-ionone, ß-ionone, and dihydro-ß-ionone, compounds that are important in herbivore-plant communication. For example, ß-ionone is part of an induced defense in canola, Brassica napus, and is released following wounding by herbivores. The objectives of the research were to evaluate whether these volatile compounds would: 1) be released in higher quantities from plants through the over-expression of the carotenoid cleavage dioxygenase1 (CCD1) gene and 2) cause herbivores to be repelled or attracted to over-expressing plants relative to the wild-type. In vivo dynamic headspace collection of volatiles coupled with gas chromatography-mass spectrometry was used to determine volatile organic compounds (VOC) in the headspace of the Arabidopsis thaliana ecotype Columbia-0 (L.) over-expressing the AtCCD1 gene. The analytical method allowed the detection of ß-ionone in the Arabidopsis headspace where emission rates ranged between 2 and 5-fold higher compared to the wild type, thus corroborating the in vivo enhancement of gene expression. A two chamber choice test between wild type and AtCCD1 plants revealed that crucifer flea beetle Phyllotreta cruciferae (Goeze) adults were repelled by the AtCCD1 plants with the highest transcription and ß-ionone levels. α-Ionone and dihydro-ß-ionone were not found in the headspace analysis, but solutions of the three compounds were tested in the concentration range of ß-ionone found in the Arabidopsis headspace (0.05 to 0.5 ng/µl) in order to assess their biological activity with crucifer flea beetle, two spotted spider mite Tetranychus urticae (Koch), and silverleaf whiteflies Bemisia tabaci (Gennadius). Choice bioassays demonstrated that ß-ionone has a strong repellent effect toward both the flea beetle and the spider mite, and significant oviposition deterrence to whiteflies. In contrast, dihydro-ß-ionone had attractant properties, especially to the crucifer flea beetle, while α-ionone did not show any significant activity. These findings demonstrate how regulating genes of the carotenoid pathway can increase herbivore deterrent volatiles, a novel tool for insect pest management.

Dillapiol: a pyrethrum synergist for control of the Colorado potato beetle.

Dillapiol, the main constituent in dill Anethum sowa Roxb. ex Fleming (Apiaceae) oil and wild pepper, Piper aduncum L. (Piperaceae), is an effective cytochrome P450 inhibitor similar to piperonylbutoxide (PBO). Laboratory and field trials with pyrethrum Chrysanthemum cinerariifolium (Trevir.) vis. extracts combined with dillapiol (1:5 and 1:16 ratio) were effective against both insecticide-susceptible and -resistant Colorado potato beetle Leptinotarsa decemlineata (Say). In the laboratory, pyrethrum efficacy was increased 2.2-fold with the SS strain and 9.1-fold with the RS strains by using pyrethrum + dillapiol. Two field trials with the pyrethrum + dillapiol formulation demonstrated efficacy > or = 10 times than that of pyrethrum alone. The residual activity (half-life) of the combination exposed to direct sunlight was 3 h but it increased to 10.7 h by adding 2% of the sunscreen octylmethoxycinnamate.

Merits of random forests emerge in evaluation of chemometric classifiers by external validation.

Real-world applications will inevitably entail divergence between samples on which chemometric classifiers are trained and the unknowns requiring classification. This has long been recognized, but there is a shortage of empirical studies on which classifiers perform best in 'external validation' (EV), where the unknown samples are subject to sources of variation relative to the population used to train the classifier. Survey of 286 classification studies in analytical chemistry found only 6.6% that stated elements of variance between training and test samples. Instead, most tested classifiers using hold-outs or resampling (usually cross-validation) from the same population used in training. The present study evaluated a wide range of classifiers on NMR and mass spectra of plant and food materials, from four projects with different data properties (e.g., different numbers and prevalence of classes) and classification objectives. Use of cross-validation was found to be optimistic relative to EV on samples of different provenance to the training set (e.g., different genotypes, different growth conditions, different seasons of crop harvest). For classifier evaluations across the diverse tasks, we used ranks-based non-parametric comparisons, and permutation-based significance tests. Although latent variable methods (e.g., PLSDA) were used in 64% of the surveyed papers, they were among the less successful classifiers in EV, and orthogonal signal correction was counterproductive. Instead, the best EV performances were obtained with machine learning schemes that coped with the high dimensionality (914-1898 features). Random forests confirmed their resilience to high dimensionality, as best overall performers on the full data, despite being used in only 4.5% of the surveyed papers. Most other machine learning classifiers were improved by a feature selection filter (ReliefF), but still did not out-perform random forests.

The effect of a synergistic concentration of a Piper nigrum extract used in conjunction with pyrethrum upon gene expression in Drosophila melanogaster.

An ethyl acetate extract of Piper nigrum L. (Piperaceae) peppercorns was tested as a synergist for the botanical insecticide pyrethrum. A high synergist ratio of 11.6 against Drosophila melanogaster was obtained for the combination of pyrethrum supplemented with P. nigrum. The effect of this combination was investigated using cDNA microarray analysis of gene expression profiles in D. melanogaster. Treatment of D. melanogaster with pyrethrum alone resulted in a large number of differentially expressed genes, principally associated with stress responses. Seven genes were identified as being commonly expressed in D. melanogaster treated with at least two of the following treatments: P. nigrum, pyrethrum or P. nigrum plus pyrethrum. These are likely implicated in Drosophila defence responses to toxins.

Efficacy of botanical insecticides from Piper species (Piperaceae) extracts for control of Ruropean chafer (Coleoptera: Scarabaeidae).

Biopesticides, including botanicals, can offer a safe and effective alternative to conventional insecticides for controlling major insect pests within an integrated pest management program. The current study highlights the practical application of a botanical insecticide for controlling a major insect pest of turfgrass: European chafer, Rhizotrogus majalis (Razoumowsky). Greenhouse and field trials were conducted to test the efficacy of a botanical formulation based on black pepper, Piper nigrum L. (Piperaceae), seed extracts to R. majalis larvae. The 7-d P. nigrum extract LC50 for R. majalis third instars was 2.5%. Successful treatment in the field was accomplished with the application of a 2% P. nigrum formulation to turfgrass infested with R. majalis second and third instars, whereas 4% extract was required in a second field trial with older third instars. The 2% pepper extract activity was comparable with the conventional insecticide diazinon in the first field trial. However, the 4% pepper extracts significantly affected the earthworm populations in treated plots compared with diazinon in the second field trail. The analysis of soil residues for piperamides in the P. nigrum extract determined a half-life of 1 - 2.6 d in the first and second field trials, respectively. This confirmed the expectation that under field conditions the residual activity would be less than conventional insecticides, thereby reducing the environmental risk associated with pesticide use. We recommend the pepper formulation for spot treatment applications when population densities reveal an epicenter of infestation rather than broadcasting over large areas, thus helping to minimize cost and negative affects on nontarget invertebrates.

Efficacy of Piper (Piperaceae) extracts for control of common home and garden insect pests.

Extracts from three species of the plant family Piperaceae, Piper nigrum [L.], Piper guineense [Schum & Thonn, and Piper tuberculatum [Jacq.], were tested for efficacy against insects from five orders. All three species contain isobutyl amides, plant secondary compounds that act as neurotoxins in insects. These materials are considered safe to mammals because Piper spp. were used for centuries for spice and medicinal purposes. When 24-h P. nigrum LC50 values were compared between common insect pests from eastern Canada and the northeastern United States, the most sensitive species in order of increasing lethal concentration were eastern tent caterpillar, Malacosoma americanum (F.) < European pine sawfly larvae, Neodiprion sertifer (Geoffroy) < spindle ermine moth larvae, Yponomeuta cagnagella [Hübner] < viburnum leaf beetle larvae, Pyrrhalta viburni [Paykull] < stripped cucumber beetle adults, Acalymma vittatum (F.) < Colorado potato beetle adults, Leptinotarsa decemlineata (Say) < Japanese beetle adults, Popillia japonica [Newman] < hairy chinch bug, Blissus leucopterus hirtis [Montandon]. The life stage tested was the point at which each species causes the greatest amount of damage to the host plant and the point at which most gardeners would likely choose to treat with a conventional synthetic insecticide. Greenhouse trials revealed that the pepper formulations also had a repellent activity, thus protecting plant leaves from 1) herbivory (lily leaf beetle, Lilioceris lilii [Scopoli], adults and larvae and stripped cucumber beetle adults) and 2) oviposition [European corn borer, Ostrinia nubilalis (Hübner)]. Combinations with other botanical extracts were additive at best in toxicity and repellent trials. Nontarget toxicity to beneficial invertebrates is a possibility because the P. nigrum LC50 for beneficial ladybird beetles was 0.2%. P. nigrum extracts can provide a reasonable level of control against lepidopteran and European pine sawfly larvae and also will work as a short-term repellent and feeding deterrent. It is recommended that the use of Piper extracts be restricted to small-scale spot treatments in residential areas where insect pest outbreaks have occurred.

Modification of senescence in ryegrass transformed with IPT under the control of a monocot senescence-enhanced promoter.

We report here the genetic modification of ryegrass senescence. Embryogenic cell suspensions of Lolium multiflorum were transformed by microprojectile bombardment with plasmid constructs containing 1.98 kb of the 5' flanking sequence of SEE1 (a maize cysteine protease gene showing enhanced expression during senescence) fused either to the Agrobacterium tumefaciens cytokinin biosynthesis gene IPT (designated PSEE1::IPT) or to the beta-glucuronidase reporter gene UIDA (PSEE1::UIDA). Plants were regenerated under selection for the HPH hygromycin resistance gene in the vector. PSEE1::UIDA transformants confirmed that the SEE1 flanking sequence functioned as a senescence-enhanced promoter in ryegrass. The IPT transgene was detected in 28 regenerants (PSEE1::IPT) from five independent transformation events. PSEE1::IPT leaves displayed a stay-green phenotype. Some PSEE1::IPT lines developed spontaneous lesions.

Botanical insecticides for controlling agricultural pests: piperamides and the Colorado Potato Beetle Leptinotarsa decemlineata say (Coleoptera: Chrysomelidae).

The efficacy of extracts from two Piperaceae species, Piper nigrum L. and P. tuberculatum Jacq. were evaluated using larvae and adults of the Colorado Potato Beetle Leptinotarsa decemlineata (Say). Young larvae and neonates were the most susceptible; a 24-h LD(50) of 0.064% extract of P. tuberculatum was determined for 4-day-old larvae, while 0.05% extract of P. nigrum reduced larval survival up to 70% within one week after treatment of potato Solanum tuberosum L. (Solanaceae) plants. When an insecticide resistant strain of L. decemlineata larvae was tested with the P. tuberculatum extract, there was less than a 2-fold tolerance ratio compared to the 22-fold tolerance ratio to cypermethrin, a pyrethroid. Older larvae, pre-pupal stage and adults, were less sensitive to the P. nigrum extracts; the 24-h LD(50) was 0.5% (95% C.I. = 0.36, 0.65). However, the same concentration was equally effective under field conditions. In the greenhouse, P. nigrum at 0.5% was as effective at reducing adult L. decemlineata feeding as combinations with 2 separate botanical mixtures, garlic and lemon grass oil. Under field conditions, the residual activity of the P. nigrum extracts was less than 3 h. When adult L. decemlineata were placed on treated plants exposed to full sunlight for 0, 1.5, and 3 h, leaf damage progressively increased as the main active compound, piperine, was found to degrade by 80% after 3 h. An in vitro polysubstrate monoxygenase (PSMO) enzyme assay, using the substrate methoxyresorufin O-demethylation (MROD), determined that the principal P. nigrum active compound, piperine, is responsible for inhibition of that specific enzyme. The results suggest that Piper extracts could be used effectively as contact botanical insect control agents to protect potato plants from developing L. decemlineata larvae at concentrations less than 0.1%. There is also potential for Piper extracts to control insecticide resistant populations in conjunction with other integrated pest management (IPM) strategies used in conventional and organic agriculture.

Improving appearance model matching using local image structure.

We show how non-linear representations of local image structure can be used to improve the performance of model matching algorithms in medical image analysis tasks. Rather than represent the image structure using intensity values or gradients, we use measures that indicate the reliability of a set of local image feature detector outputs. These features are image edges, corners, and gradients. Feature detector outputs in flat, noisy regions tend to be ignored whereas those near strong structure are favoured. We demonstrate that combinations of these features give more accurate and reliable matching between models and new images than modelling image intensity alone. We also show that the approach is robust to non-linear changes in contrast, such as those found in multi-modal imaging.

Inhibition of glutathione S-transferases (GSTs) from parasitic nematodes by extracts from traditional Nigerian medicinal plants.

Piliostigma thonningii, Ocimum gratissimum, Nauclea latifolia and Alstonia boonei are used in Nigerian traditional medicines against gastrointestinal helminths of animals and man. Proanthocyanidins were detected in Piliostigma and Nauclea, but not Alstonia or Ocimum. Extracts of these plants killed 50% of brine shrimp nauplii at <10 ppm (Nauclea), 100 ppm (Piliostigma) and <1000 ppm (Ocimum and Alstonia), the Nauclea LD50 being similar to the anthelmintic drug piperazine. Extracts were also toxic to the parasitic nematode Haemonchus infective L3 stage. Nematode glutathione-S-transferases (GSTs) are potential drug targets. Apart from Alstonia all the medicinal plants contained heat-stable inhibitory activities against recombinant Ascaris and Onchocerca GSTs in vitro. Piliostigma, Ocimum and Nauclea had IC50s of 2, 10 and 15 microg/mL respectively for Ascaris GST and 4, 8, 28 microg/mL respectively for Onchocerca GST. We suggest that the inhibitory properties of some of these Nigerian plant extracts against GST may contribute to the pharmacological basis of their efficacy against helminths in traditional herbal use.

Characterisation of a cysteine protease cDNA from Lolium multiflorum leaves and its expression during senescence and cytokinin treatment.

A cysteine protease cDNA clone (See1) highly homologous to barley aleurain was isolated from Lolium multiflorum leaves. During leaf senescence, expression of the See1 mRNA and protein was strongly enhanced. In dark-incubated leaf segments, cytokinin delayed senescence and reduced expression of both See1 mRNA and protein.

The toxicity of a neem insecticide to populations of culicidae and other aquatic invertebrates as assessed in in situ microcosms.

Microcosm trials were conducted with the botanical insecticide Margosan-O(R) to assess the potential hazards of the product to aquatic organisms. Laboratory chronic bioassays with water from the treated microcosms were conducted to provide an estimate of the residual effect of Margosan-O. Results from chronic tests showed Margosan-O toxicity to be greater in the laboratory exposures than in situ with Culicidae larvae exposed to the same concentrations. Residue analyses of the active ingredient, azadirachtin, determined that it had a half-life of 36 to 48 h in water exposed to natural sunlight. Two applications of Margosan-O at the recommended application rate for pests did not harm aquatic invertebrates that are categorized as planktonic and filter feeding (Culex sp. and Daphnia sp.). However, the benthic invertebrate (Chironomus riparius) was affected by multiple applications of neem. These results show that the use of Margosan-O and possibly other neem extracts in or near aquatic environments could lead to disturbances in benthic populations and may cause decreases in numbers of organisms that are important in food web and nutrient cycling processes.

Expression of the abnormal gravitropism phenotypes creep and ageotropum during development in pea.

This study established that the mutant creep and ageotropum phenotypes are expressed differently during development in pea (Pisum sativum L.). Etiolated ageotropum stems grew at a wide range of angles, whereas etiolated creep stems emerged vertically from the compost. However, when etiolated creep stems were subjected to additional gravitropic demands, such as growth to excessive height or reorientation, abnormality was detectable. When plants were handled to a greater extent, earlier loss of vertical growth resulted. In light-grown shoots, creep lost vertical orientation, whereas ageotropum grew more normally. Root systems of creep exhibited normal growth patterns, whereas ageotropum main and lateral roots grew at abnormal angles. Thus, the ageotropum mutation strongly affects gravitropism in roots and etiolated stems, whereas the creep mutation affects both light- and dark-grown stems, but is most apparent in older, taller plants.

Expression of the UNC-5 guidance receptor in the touch neurons of C. elegans steers their axons dorsally.

Growth cones in developing nervous systems encounter a sequence of extracellular cues during migration. In theory, a growth cone can navigate by selectively expressing or activating surface receptor(s) that recognize extracellular cues appropriate to each migratory phase. Using the simple Caenorhabditis elegans nervous system, we attempted to demonstrate that path selection by migrating growth cones can be predictably altered by ectopic expression of a single receptor. The unc-5 gene of C. elegans encodes a unique receptor of the immunoglobulin superfamily (UNC-5), required cell-autonomously to guide growth cone and mesodermal cell migrations in a dorsal direction on the epidermis. We report here that the UNC-5 receptor induces dorsally oriented axon trajectories when ectopically expressed in the touch receptor neurons which normally extend pioneer axons longitudinally or ventrally on the epidermis. These errant trajectories depend on unc-6, which encodes a putative epidermal path cue, just as normal dorsally oriented axon trajectories do (such as those of certain motor neurons), suggesting that UNC-5 acts to reorient the touch cell growth cones by using its normal guidance mechanisms. These results support previous evidence that UNC-5 and UNC-6 play instructive rules in guiding growth cone migrations on the epidermis in C. elegans, and indicate that pioneering growth cones, which normally migrate in different directions, may use equivalent intracellular signalling mechanisms for guidance.

The mec-7 beta-tubulin gene of Caenorhabditis elegans is expressed primarily in the touch receptor neurons.

Mutants of the mec-7 beta-tubulin gene of Caenorhabditis elegans lack the large diameter 15-protofilament microtubules normally found only in the set of six touch receptor neurons. Both a mec-7-lacZ reporter gene and affinity-purified anti-mec-7 antibodies were used to show that mec-7 is expressed primarily in the touch neurons. These data are consistent with a possible instructive role for the mec-7 tubulin in determining microtubule protofilament number. The antibodies and the mec-7-lacZ transgene were also used to examine mec-7 expression in mutants affecting the generation, differentiation or maintenance of the touch neurons. Decreased expression was observed in mutants of unc-86 and mec-3, genes that encode transcription factors essential for touch receptor neuron generation and differentiation, respectively.

Genetic and molecular analysis of vgU and vgW: two dominant vg alleles associated with gene fusions in Drosophila.

In the absence of a vg+ gene, extensive cell death occurs in third instar imaginal discs, which results in a complete loss of adult wing margin structures. Essentially all molecularly characterized vg alleles are associated with deletions or insertions of DNA into the vg locus. These alterations reduce or eliminate a 3.8-kb vg-specific transcript, resulting in recessive loss of function alleles. We report here the analysis of two dominant vg alleles which have been identified (vgU and vgW). The vgU allele is associated with a chromosomal inversion which splits the vg locus, resulting in a gene fusion between vg and the mastermind (mam) neurogenic locus. Reversion analysis of vgU indicates that sequences from the mam locus are required for vgU dominance. The vgW allele is also the result of a chromosomal inversion, in this case resulting in a gene fusion between vg and the homeobox-containing invected (inv) gene. It is also associated with novel dominant homeotic transformations. Revertant analysis indicates that sequences from inv are required for the dominant wing and dominant homeotic effects of vgW. The vg dominance does not appear to be mediated through a reduction of vg expression or a novel fusion transcript in either vgU or vgW. The results are consistent with a model in which inappropriate expression of inv causes the dominant homeotic effects seen in vgW.

Cellular basis of the effects of gibberellin and the pro gene on stem growth in tomato.

Tomato (Lycopersicon esculentum Mill.) plants homozygous for the mutant pro gene, exhibiting the distinctive procera phenotype, appeared virtually identical to gibberellic acid (GA3)-treated isogenic normal plants. The pro gene and GA3 caused analogous increases in internode length, and in the length and number of cells in the outer cell layers of each internode. Internode number was also increased by pro and GA3 over the period of the experiment. Despite their greater length, the internodes of GA3-treated and pro plants reached their final size within a time period similar to that of internodes of untreated normal plants. The pro mutant itself was responsive to GA3, especially in the seedling stage, but the proportional increase in height seen in the later stages of growth was less than that of normal plants.

Apical localization of 1-aminocyclopropane-1-carboxylic acid and its conversion to ethylene in etiolated pea seedlings.

The biosynthetic basis for the high rates of ethylene production by the apical region of etiolated pea (Pisum sativum L.) seedlings was investigated. The ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) was quantified in extracts of various regions of seedlings by measuring isotopic dilution of a (2)H-labelled internal standard using selected-ion-monitoring gas chromatography/mass spectrometry. The ACC levels in the apical hook and leaves were much higher than in the expanded internodes of the epicotyl. The capacity of excised tissue sections to convert exogenous ACC to ethylene was also much greater in the apical region, reflecting the distribution of soluble protein in the epicotyl.

Amyloplast development in etiolated and ethylene-treated pea epicotyls.

The amyloplasts found in the apical hook cells of etiolated pea (Pisum sativum L.) epicotyls were randomly distributed. Sedimentation of endodermal amyloplasts in the direction of gravity became apparent in the transition from the hook to the top of the main axis of the epicotyl. Cortical amyloplasts in this region were not, however, sedimented. These patterns of sedimentation could not be related to changes in amyloplast size, and it is proposed that cytoplasmic properties determine amyloplast behaviour.The differentiation of plastids in the hook differed between the amyloplast-containing endodermal cells and the cortical cells, in which amoeboid plastids predominated over amyloplasts. Amyloplasts disappeared from the cortical cells in the main axis of the epicotyl, but in the endodermal cells sedimented amyloplasts were found throughout the upper epicotyl.Etiolated epicotyls induced to grow horizontally by treatment with ethylene had a normal content of amyloplasts, sedimented in the direction of gravity.