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This article discusses the treatment of erythromelanosis follicularis faciei et colli (EFFC) with a 595-nm pulsed dye laser (PDL). EFFC is a condition characterized by telangiectasia, hyperpigmentation, and follicular papules on the face and neck, with a higher prevalence among individuals of Asian ancestry. Current treatments for EFFC are limited in efficacy, but this study presents three cases where treatment with the 595-nm PDL resulted in significant improvement. The PDL works by targeting blood vessels and melanin, and its integration of dynamic cooling technology reduces the risk of post-treatment pigmentation. However, the mechanism by which the PDL improves follicular plugging is not yet clear. The study demonstrates the clinical effectiveness and safety of the PDL treatment for EFFC, but further research is needed due to the small number of patients involved. [Extracted from the article]

CQMUH‐011 is a modified adamantane sulfonamide compound, that inhibits macrophage proliferation and possesses anti‐inflammatory properties. Here, fresh mouse splenocytes were obtained and stimulated with concanavalin A (ConA, 5 μg/ml) in vitro; and experimental autoimmune hepatitis (AIH) was induced by ConA (20 mg/kg, iv) in vivo, to clarify the protective effects of CQMUH‐011 against AIH and its possible mechanisms. Our results demonstrated that CQMUH‐011 pretreatment can dose‐dependently inhibit the proliferation of splenocytes in vitro. In vivo, CQMUH‐011 administration reduced the hepatic histopathological score and the infiltration of lymphocytes in the liver parenchyma; additionally, it downregulated the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and pro‐inflammatory cytokines interferon (IFN)‐γ, tumor necrosis factor (TNF)‐α, and interleukin (IL)‐6 in serum, as well as those of methane dicarboxylic aldehyde and myeloperoxidase in the liver tissues. It also down‐regulated the expression of p‐NF‐κB and related proteins in the liver tissues. Furthermore, CQMUH‐011 could maintain the balance of CD3+CD4+/CD3+CD8+ and decrease the percentages of CD8+CD69+ and CD4+CD25+/‐CD69+ T‐cells in the splenocytes of ConA‐challenged mice. Moreover, we found thatCD4+CD25+/−CD69+ T‐cells were significantly correlated with ALT levels, especially CD4+CD25−CD69+ T‐cells. In conclusion, CQMUH‐011 exerts potential protective effects against ConA‐induced hepatitis, which may be partially attributed to its inhibition of T cells, especially the suppression of the proliferation of CD4+CD25−CD69+ and CD8+CD69+ subsets in the spleen. CQMUH‐011 also reduced the early apoptosis of lymphocytes in the thymus. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the functions of miR-214-3p in diabetic neuropathic rodents. The diabetic neuropathy was induced by intraperitoneal injection of streptozotocin (STZ) in rats, and miR-214-3p was delivered via tail vein injection of lentivirus. Hot or cold stimulus tests demonstrated that STZ induced thermal hyperalgesia. Neurophysiological measurements revealed that motor and sensory nerve conduction velocity and nerve blood flow were decreased in diabetic neuropathic rats. However, the STZ-induced hyperalgesia, and reduced nerve conduction velocity and nerve blood flow were all significantly reversed by miR-214-3p administration. HE staining, TUNEL, ELISA, and immunoblotting demonstrated that STZ led to obvious pathological lesion, cell apoptosis, and inflammation in dorsal root ganglion (DRG), evidenced by altered levels of apoptosis-related protein molecules and inflammatory factors, and activation of Toll-like receptor 4 (TLR4)/myeloid differentiation primary response gene 88/nuclear factor kappa B signaling. The pathological alterations in diabetic neuropathic rats in DRG were significantly ameliorated by miR-214-3p application. In addition, sodium channel protein type 3 subunit alpha isoform 1 (Nav1.3) and TLR4 were identified as targets of miR-214-3p via dual-luciferase reporter assay. MiR-214-3p may play its roles by downregulating Nav1.3 and TLR4. In summary, miR-214-3p alleviated diabetes-induced nerve injury, and pathological lesion, cell apoptosis, and inflammation in DRG by regulating Nav1.3 and TLR4 in STZ-induced rats. These findings may provide novel therapeutic targets for clinical treatment of diabetic neuropathy. [ABSTRACT FROM AUTHOR]

Male moths possess highly sensitive and selective olfactory systems that detect sex pheromones produced by their females. Pheromone receptors (PRs) play a key role in this process. The PR HassOr14b is found to be tuned to (Z)-9-hexadecenal, the major sex-pheromone component, in Helicoverpa assulta. HassOr14b is co-localized with HassOr6 or HassOr16 in two olfactory sensory neurons within the same sensilla. As HarmOr14b, the ortholog of HassOr14b in the closely related species Helicoverpa armigera, is tuned to another chemical (Z)-9-tetradecenal, we study the amino acid residues that determine their ligand selectivity. Two amino acids located in the transmembrane domains F232I and T355I together determine the functional difference between the two orthologs. We conclude that species-specific changes in the tuning specificity of the PRs in the two Helicoverpa moth species could be achieved with just a few amino acid substitutions, which provides new insights into the evolution of closely related moth species. [ABSTRACT FROM AUTHOR]

The low-boom inverse design method is of great interest for the preliminary design of supersonic transport (SST), owing to its good capability of reducing sonic boom at low computational cost. However, the main challenge is how to prescribe an attainable target. This paper proposes an inverse design method using a new target-generation strategy to address this challenge. Unlike existing methods such as the Jones-Seebass-George-Darden method and the mixed-fidelity inverse design method [the Li-Shields-Geiselhart (LSG) method], the proposed method directly parameterizes the near-field overpressure distribution of a reference configuration and minimizes the ground perceived level in decibels (PLdB) predicted by solving the augmented Burgers equation to obtain an optimal target distribution, subject to cruise lift and volume constraints. The proposed method is demonstrated by the Japan Aerospace Exploration Agency (JAXA) Wing Body case, which shows that it mitigates sonic boom under lift constraint and achieves a reduction of 2.07 PLdB more than the LSG method. The proposed method is applied to the low-boom design of a large SST configuration called NPU-T7104, which is close in size to the Tupolev Tu-144; results show that aerodynamic performance is maintained while shock wave coalescence is effectively prevented, and ground-boom intensity is dramatically reduced. [ABSTRACT FROM AUTHOR]

Helicoverpa armigera and H. assulta are sympatric closely related species sharing two sex pheromone components, (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-9-hexadecenal (Z9-16:Ald) but in opposite ratios, 97:3 and 3:97 respectively. This feature makes them a feasible model for studying the evolution of pheromone coding mechanisms of lepidopteran insects. Despite a decade-long study to deorphanize the pheromone receptor (PR) repertoires of the two species, the comparison of the function of all PR orthologs between the two species is incomplete. Moreover, the ligands of OR14 and OR15 have so far not been found, likely due to the missing of the active ligand(s) in the compound panel and/or incompatibility of heterologous expression systems used. In the present study, we expressed the PR repertoires of both Helicoverpa species in Drosophila T1 neurons to comparatively study the function of PRs. Among those PRs, OR13, OR6, and OR14 of both species are functionally conserved and narrowly tuned, and the T1 neurons expressing each of them respond to Z11-16:Ald, (Z)-9-hexadecenol (Z9-16:OH), and (Z)-11-hexadecenyl acetate (Z11-16:Ac), respectively. While HarmOR16-expressing neurons respond strongly to (Z)-9-tetradecenal (Z9-14:Ald) and (Z)-11-hexadecenol (Z11-16:OH), the neurons expressing HassOR16 mainly respond to Z9-14:Ald and also weakly respond to (Z)-9-tetradecenol (Z9-14:OH). Moreover, HarmOR14b-expressing neurons are activated by Z9-14:Ald, whereas HassOR14b-expressing neurons are sensitive to Z9-16:Ald, Z9-14:Ald, and (Z)-9-hexadecenol (Z9-16:OH). In addition, HarmOR15-expressing neurons are selectively responsive to Z9-14:Ald. However, the Drosophila T1 neurons expressing either HarmOR11 or HassOR11 are silent to all of the compounds tested. In summary, except for OR11, we have deorphanized all the PRs of these two Helicoverpa species using a Drosophila expression system and a large panel of pheromone compounds, thereby providing a valuable reference for parsing the code of peripheral coding of pheromones. [Display omitted] • PRs of H. armigera and H. assulta were deorphanized in Drosophila T1 neurons. • OR13, OR6 and OR14 respond to Z11-16:Ald, Z9-16:OH and Z11-16:Ac, respectively. • HarmOR16 responds to Z9-14:Ald and Z11-16:OH, HassOR16 to Z9-14:Ald and Z9-14:OH. • OR14b is the receptor tuned to Z9-16:Ald in H. assulta , but is not in H. armigera. • HarmOR15 responds strongly to Z9-14:Ald and weakly to Z9-14:OH. [ABSTRACT FROM AUTHOR]

The endoparasitoid wasp Campoletis chlorideae Uchida attacks many noctuid species which vary in their suitability but the host association of this wasp species is poorly understood. In this study, three sympatric noctuid species were chosen, Helicoverpa armigera (Hübner), Mythimna separata (Walker), and Spodoptera exigua (Hübner), to study host preference and suitability of C. chlorideae, examining the immunological compatibility between the parasitoid and the three host species., Campoletis chlorideae parasitised all three species, but showed a much higher preference for H. armigera and M. separata than for S. exigua. In H. armigera, the young wasps developed perfectly and none was encapsulated. In M. separata, about 50% and 10% of them were encapsulated in single and double parasitisation experiments, respectively, and the cocoons were significantly lighter than those from parasitised H. armigera and S. exigua. In S. exigua, 84.8% of the young wasps were encapsulated., In double parasitisation experiments, the phenoloxidase activity, the number of total haemocytes, and the number of plasmatocytes and granulocytes significantly decreased in the haemolymph of H. armigera and M. separata, but not in S. exigua., These results indicate that H. armigera is the most suitable host, M. separata is moderately suitable, whereas S. exigua is an unsuitable host for C. chlorideae. The suitability of the parasitoid to three host species was closely related with the capacity of the wasp to inhibit the host immune system., This study sustains the optimality theory and also evidences the correlation of host-selection behaviour of the parasitoid wasp and its immunosuppressive ability. [ABSTRACT FROM AUTHOR]

Odorant-binding proteins (OBPs) are soluble proteins, whose role in olfaction of insects is being recognized as more and more important. We have cloned, expressed and purified an OBP (HarmOBP7) from the antennae of the moth Helicoverpa armigera. Western blot experiments indicate specific expression of this protein in the antennae of adults. HarmOBP7 binds both pheromone components Z-11-hexadecenal and Z-9-hexadecenal with good affinity. We have also performed a series of binding experiments with linear aldehydes, alcohols and esters, as well as with other compounds and found a requirement of medium size for best affinity. The affinity of OBP7, as well as that of a mutant lacking the last 6 residues does not substantially decrease in acidic conditions, but increases at basic pH values with no significant differences between wild-type and mutant. Binding to both pheromone components, instead, is negatively affected by the lack of the C-terminus. A second mutant, where one of the three lysine residues in the C-terminus (Lys123) was replaced by methionine showed reduced affinity to both pheromone components, as well as to their analogues, thus indicating that Lys123 is involved in binding these compounds, likely forming hydrogen bonds with the functional groups of the ligands. [ABSTRACT FROM AUTHOR]

Abstract: The two sibling species Helicoverpa armigera and Helicoverpa assulta utilise the same two aldehydes as their sex pheromones, but in opposite ratios. In both species three odorant-binding proteins (OBPs) can be classified as pheromone-binding proteins (PBPs). To investigate the role of these three PBPs in chemical communication between sexes and their mode of action, we have expressed the proteins in bacteria and prepared mutants lacking their C-terminal regions. Using polyclonal antibodies we found that the expression of the three PBPs is basically confined to the antennae of both sexes and both species. Binding experiments with the fluorescent probe N-phenyl-1-naphthylamine across a pH range indicated that, the affinity of wild-type proteins decreases at low pH, while that of the mutants is not or less affected, suggesting that a conformational change of the C-terminus occurs in these proteins, as reported for other lepidopteran OBPs. All three proteins bind with similar strength both pheromone components, as well as their corresponding alcohols and acetates. However, they exhibit significant selectivity to linear alcohols and aldehydes of different length, with optimal affinities to the ligand of 13–15 carbon atoms for PBP1 and 12–14 carbon atoms for PBP2. We suggest that all three PBPs might cooperate to build a unique olfactory image, that could help avoiding cross-mating between the two species and with other noctuids. [Copyright &y& Elsevier]

Odorant-binding proteins (OBPs) mediate both perception and release of semiochemicals in insects. These proteins are the ideal targets for understanding the olfactory code of insects as well as for interfering with their communication system in order to control pest species. The two sibling Lepidopteran species Helicoverpa armigera and H. assulta are two major agricultural pests. As part of our aim to characterize the OBP repertoire of these two species, here we focus our attention on a member of this family, OBP10, particularly interesting for its expression pattern. The protein is specifically expressed in the antennae of both sexes, being absent from other sensory organs. However, it is highly abundant in seminal fluid, is transferred to females during mating and is eventually found on the surface of fertilised eggs. Among the several different volatile compounds present in reproductive organs, OBP10 binds 1-dodecene, a compound reported as an insect repellent. These results have been verified in both H. armigera and H. assulta with no apparent differences between the two species. The recombinant OBP10 binds, besides 1-dodecene, some linear alcohols and several aromatic compounds. The structural similarity of OBP10 with OBP1 of the mosquito Culex quinquefasciatus, a protein reported to bind an oviposition pheromone, and its affinity with 1-dodecene suggest that OBP10 could be a carrier for oviposition deterrents, favouring spreading of the eggs in these species where cannibalism is active among larvae. [ABSTRACT FROM AUTHOR]

Abstract: Helicoverpa assulta and Helicoverpa armigera are sibling species with different host-plant ranges. We have previously reported electrophysiological and behavioral responses of H. armigera to sugars and amino acids. Here we describe a parallel study performed on H. assulta and compare the results obtained with the two species. In females, fourteen gustatory chemosensilla, identified on one ventrolateral side of the fifth tarsomere were stimulated with sucrose, glucose, fructose, maltose, myo-inositol, and the twenty common amino acids, using the tip-recording technique. The taste receptor neurons in eight chemosensilla were identified sensitive to the sugars, myo-inositol, Lys, Glu, Arg, Trp, and Ser which all induced proboscis extension reflex (PER) when tarsi were stimulated. There was a positive correlation between electrophysiological activities and PER responses triggered by sucrose. No stimulatory effect on oviposition was observed with sugar or amino acid mixtures. In males, three chemosensilla showed responses to the four sugars, but generally weaker than in females. The major difference of the two species was the variety of amino acids triggering electrophysiological responses. The stimulatory effect of sugars and amino acids on H. assulta was also generally weaker than that on H. armigera. [Copyright &y& Elsevier]

This Technical Note is the first description of a large-scale logarithmic flow-rate damping system designed to retain cells of different adherence, different suspensibility and different motility. The chamber, which can easily retain and cultivate many types of cells, including high-motility cells and swimming cells, viaa series of “speed bumps”, readily facilitates cell retention for complex heterogeneous cultures. Yeast cells, red blood cells, rabbit bone marrow aspirate and dinoflagellate swimming cells were introduced into the chip for multi-cell retention, multi-cell culture and observation. Here, we show that the chamber creates a flow field with a ratio of end/start speeds as low as 0.01. The logarithmic distribution of flow-rate within the chamber is controlled precisely by pressure, all of the cell types that we tested were retained easily within the chamber. Many cell–cell interactions were observed, predicting a high potential for the success of on-chip heterogeneous cell experiments. [ABSTRACT FROM AUTHOR]

The role of IgG antibodies in inducing or modifying allergic reaction has not been sufficiently clarified. The objective of this investigation is to elucidate the relationship between IgG antibodies and penicillin allergy, between IgG and IgE antibodies in allergic patients. Enzyme-linked immunosorbent assay and Radioallergosorbent test were used to examine eight kinds of specific IgG and IgE antibodies, including major antigenic determinants: benzylpenicilloyl (BPO), ampicilloyl (APO), amoxicilloyl (AXO) and phenoxomethylpenicilloyl (PVO), and minor antigenic determinants: benzylpenicillanyl (BPA), ampicillanyl (APA), amoxicillanyl (AXA) and phenoxomethylpenicillany (PVA), in the sera of 249 patients with penicillin allergy. Except BPA-IgG, seven kinds of antigenic determinants IgG antibodies levels were significantly higher than that of control group ( P < 0.05). Positive rates of specific IgG and IgE were 47.0 and 57.8%, while positive rate of IgE and IgG together was 77.9%. The positive rate of IgG antibodies to major antigenic determinants (42.2%) was significantly higher than that of minor antigenic determinants (8.8%) ( P < 0.05). The positive rate of IgG antibodies of patients with typical clinical symptoms after penicillin administration when skin tests were negative was significantly higher than that of patients with positive skin test ( P < 0.01). There were no differences between the IgG positive rates to three kinds of determinants and that of all of eight kinds. The study indicates that IgG may be important in penicillin allergy with negative skin test and IgG antibodies to major antigenic determinants probably play a more important role in the process of allergic reaction. [ABSTRACT FROM AUTHOR]

The findings of numerous studies have suggested that both genetic and environmental influences are involved in the pathogenesis of allergic disease and atopy. We studied the polymorphisms in the interferon (IFN)-gamma (γ) and IFN-γ receptor 1 (IFNR1) gene with the aim of clarifying the relationships among these polymorphisms, penicillin allergy and anti-penicillin antibodies. A restriction endonuclease fragment length polymorphism (RFLP)-PCR analysis and sequencing were used to study the IFNR1 and IFN-γ polymorphisms. The presence and level of eight specific immunoglobulin (Ig)E and IgG antibodies were determined by the radioallergosorbent test (RAST) and enzyme-linked immunosorbent assay (ELISA), respectively. The positive rates of specific IgE and IgG were 61.11 and 53.92%, respectively. There was no significant difference in the whole-allele of IFN-γ distribution between patients with a penicillin allergy and control subjects. Allele 7 (18CA repeat) was significantly less frequent in the urticaria group (3.19 vs. 11.93%) than in the controls. There was no difference in IFN-γ production among different alleles in IFN-γ. The frequency of G/A (Val/Met) in the IFNR1 gene in allergic patients was significantly less than that in the controls ( P < 0.05). There were no significant differences in the positive rate of IgE among different alleles of IFN-γ. The same was true for the positive rate of IgG. The Met/Val allele in IFNR1 gene may have a protective role in the non-penicillin allergic population. The allele 18CA repeat in IFN-γ gene may be associated with urticaria. [ABSTRACT FROM AUTHOR]

Objective: Omeprazole, lansoprazole and rabeprazole have been widely used as proton pump inhibitors (PPIs). They can be metabolized in the liver by CYP2C19, a polymorphic enzyme, and have a wide interindividual variability with respect to drug response. In the investigation reported here, we examined the kinetic characteristics of the three PPIs in healthy Chinese subjects in relation to CYP2C19 genotype status. Methods: Six homozygous extensive metabolizers (homEMs), six heterozygous extensive metabolizers (hetEMs) and six poor metabolizers (PMs) were recruited for the study from a total of 90 healthy Chinese volunteers whose CYP2C19 genotype status was determined by means of PCR-restriction fragment length polymorphism (RFLP). The study was had an open label, randomized, three-way crossover design. After a single oral dose of 40 mg omeprazole, 30 mg lansoprazole or 40 mg rabeprazole, plasma concentrations of the three PPIs were determined by HPLC. Results: There were some differences for the area under the plasma concentration-time curve (AUC), the elimination half-life (t1/2ke) and the maximum plasma concentration (cmax) in the three groups. In the homEMs, hetEMs and PMs, the relative AUC0–∞ values were 1:2.8:7.5 for omeprazole, 1:1.7:4.0 for lansoprazole and 1:1.6:3.7 for rabeprazole, respectively; the relative t1/2ke values were 1:1.02:1.65 for omeprazole, 1:1.08:2.39 for lansoprazole and 1:1.37:1.85 for rabeprazole, respectively; the relative cmax values were 1:2.09:4.39 for omeprazole, 1:1.34:1.72 for lansoprazole, and 1:1.24:2.04 for rabeprazole, respectively. Conclusion: The pharmacokinetic characteristics of the three PPIs are significantly dependent on the CYP2C19 genotype status. These data indicate that individualized dose regimen of the three PPIs, based on identification of genotype, can be of great benefit for ensuring the reasonable use of these drugs. [ABSTRACT FROM AUTHOR]

Objective: Although IL-4 and IL-13 share many biological activities, some are unique to IL-13. We genotype the IL-13 and IL-4IL-13-SNPs genes for polymorphisms that could then be used to determine associations with IgE regulation as well as levels of IL-4 and IL-13. Methods: Eight kinds of IgE specific to penicillins were determined with radioallergosorbent test (RAST) in the sera of 158 patients with penicillin allergies and 89 healthy subjects. Serum levels of IL-4 and IL-13 were measured by using enzyme-linked immunosorbent assay (ELISA). The genotyping of IL-13 Arg130Gln, IL-4-IL-13-SNP3 and IL- 4-IL13-SNP4 was carried out by polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) technique. Results: Among patients with positive specific IgE, significant differences between the IL-4-IL-13-SNP3 and IL-4-IL-13-SNP4 genotypes were observed between patients with positive BPA and the control group (P«0.05, P«0.05). Additionally, we also found significant differences in the IL-4-IL-13-SNP4 genotype between positive and negative BPA-IgE patients (P«0.05). However, we found no significant differences in the prevalence of these polymorphisms between any of the symptomatic groups studied and the control group. Nor were there any significant differences between levels of IL-4 and IL-13 and any of genotypes. Conclusion: The data suggest that IL-4-IL-13-SNP genes between IL-4 and IL-13 play a role in the regulation of specific IgE levels in patients with penicillin allergies. [ABSTRACT FROM AUTHOR]

The tobacco cutworm Spodoptera litura is one of the most destructive polyphagous crop pests. Olfaction and taste play a crucial role in its host plant selection and sexual communication, but the expression profile of chemosensory genes remains unclear. In this study, we identified 185 chemosensory genes from 7 organs in S. litura by transcriptome sequencing, of which 72 genes were published for the first time, including 27 odorant receptors (ORs), 26 gustatory receptors (GRs), 1 ionotropic receptor (IR), 16 odorant‐binding proteins (OBPs), and 2 chemosensory proteins (CSPs). Phylogenetic analyses revealed that ORs, IRs, OBPs, and sensory neuron membrane proteins (SNMPs) were mainly expressed in antennae and sequence‐conserved among Noctuidae species. The most differentially expressed genes (DEGs) between sexes were ORs and OBPs, and no DEGs were found in GRs. GR transcripts were enriched in proboscis, and the expression of sugar receptors was the highest. Carbon dioxide receptors, sugar receptor‐SliuGR6, and bitter GRs‐SlituGR43 and SlituGR66 had higher sequence identities between Noctuidae species. CSPs were broadly expressed in various organs, and SlituCSP13 was a DEG in adult antennae. The functional analysis in the Drosophila OR67d expression system found that SlituOR50, a receptor highly expressed in female antennae, is selectively tuned to farnesyl acetate. The results provide a solid foundation for understanding the molecular mechanisms by which chemosensory genes operate to elicit behavioral responses in polyphagous insects. [ABSTRACT FROM AUTHOR]

The sympatric closely related species Helicoverpa armigera and Helicoverpa assulta use 97:3 and 7:93 of (Z)-11-hexadecenal and (Z)-9-hexadecenal, respectively, as their sex pheromone to find/locate correct sex mates. Moreover, (Z)-11-hexadecenyl alcohol and (Z)-9-hexadecenyl alcohol are more abundant in the pheromone gland of H. assulta than in that of H. armigera . To clarify the molecular basis of these differences, we sequenced the pheromone gland transcriptomes of the two species and compared the expression patterns of the candidate enzyme genes involved in the pheromone biosynthetic pathways by FPKM values and quantitative RT-PCR analysis. We found that the desaturase gene LPAQ expressed about 70 times higher in H. armigera than in H. assulta , whereas another desaturase gene NPVE expressed about 60 times higher in H. assulta than in H. armigera . We also observed significantly higher expression of the fatty acyl reductase (FAR) gene FAR1 and the aldehyde reductase (AR) gene AR3 in H. assulta than in H. armigera . Examination of the pheromone glands of the backcross offspring of their hybrids to H. assulta showed a positive linear correlation between the expression level of LPAQ and the amount of Z11-16:Ald and between the expression level of NPVE and the amount of Z9-16:Ald in the pheromone glands. Taken together, these data demonstrate that the expressional divergences of LPAQ and NPVE determine the opposite sex pheromone component ratios in the two species and the divergent expression of FAR1 and AR3 may account for the greater accumulation of alcohols in the pheromone gland of H. assulta . [ABSTRACT FROM AUTHOR]

Large numbers of neutrophils infiltrate tumors and comprise a notable component of the inflammatory tumor microenvironment. While it is established that tumor cells exhibit the Warburg effect for energy production, the contribution of the neutrophil metabolic state to tumorigenesis is unknown. Here, we investigated whether neutrophil infiltration and metabolic status promotes tumor progression in an orthotopic mouse model of pancreatic ductal adenocarcinoma (PDAC). We observed a large increase in the proportion of neutrophils in the blood and tumor upon orthotopic transplantation. Intriguingly, these tumor-infiltrating neutrophils up-regulated glycolytic factors and hypoxia-inducible factor 1-alpha (HIF-1α) expression compared to neutrophils from the bone marrow and blood of the same mouse. This enhanced glycolytic signature was also observed in human PDAC tissue samples. Strikingly, neutrophil-specific deletion of HIF-1α (HIF-1αΔNφ) significantly reduced tumor burden and improved overall survival in orthotopic transplanted mice, by converting the pro-tumorigenic neutrophil phenotype to an anti-tumorigenic phenotype. This outcome was associated with elevated reactive oxygen species production and activated natural killer cells and CD8+ cytotoxic T cells compared to littermate control mice. These data suggest a role for HIF-1α in neutrophil metabolism, which could be exploited as a target for metabolic modulation in cancer. [ABSTRACT FROM AUTHOR]

Lepidopteran caterpillars rely on olfaction and gustation to discriminate among food sources. Compared to the larval gustation, the larval olfaction has been poorly investigated. To uncover the molecular basis of olfaction in Helicoverpa armigera larvae, we identified 17 odorant receptor (Or) genes in larval antennae and maxillae using transcriptome sequencing, and functionally identified the response spectra of seven Ors to ecologically relevant odorants. Innate behavioural responses of larvae to active odorants were evaluated in chemotaxis assays. Several odorant blends were formulated based on the Ors tuning spectra and caterpillar chemotaxis. A four-component blend strongly attracted H. armigera larvae, and cis -jasmone and 1-pentanol were identified as essential components. Localization analyses showed that the two Ors detecting these components (Or41 and Or52) were expressed in the same sensory neurons. This is the first evidence that Ors in a polyphagous caterpillar respond to odorants in a combinatorial manner. The design of attractants to target specific olfactory pathways may promote the development of new baits for pest management. [ABSTRACT FROM AUTHOR]

Sex pheromones are pivotal for insect reproduction. However, the mechanism of sex pheromone communication remains enigmatic in hymenopteran parasitoids. Here we have identified the sex pheromone and elucidated the olfactory basis of sex pheromone communication in Campoletis chlorideae (Ichneumonidae), a solitary larval endoparasitoid of over 30 lepidopteran pests. Using coupled gas chromatography-electroantennogram detection, we identified two female-derived pheromone components, tetradecanal (14:Ald) and 2-heptadecanone (2-Hep) (1:4.6), eliciting strong antennal responses from males but weak responses from females. We observed that males but not females were attracted to both single components and the blend. The hexane-washed female cadavers failed to arouse males, and replenishing 14:Ald and 2-Hep could partially restore the sexual attraction of males. We further expressed six C. chlorideae male-biased odorant receptors in Drosophila T1 neurons and found that CchlOR18 and CchlOR47 were selectively tuned to 14:Ald and 2-Hep, respectively. To verify the biological significance of this data, we knocked down CchlOR18 and CchlOR47 individually or together in vivo and show that the attraction of C. chlorideae to their respective ligands was abolished. Moreover, the parasitoids defective in either of the receptors were less likely to court and copulate. Finally, we show that the sex pheromone and (Z)-jasmone, a potent female attractant, can synergistically affect behaviors of virgin males and virgin females and ultimately increase the parasitic efficiency of C. chlorideae. Our study provides new insights into the molecular mechanism of sex pheromone communication in C. chlorideae that may permit manipulation of parasitoid behavior for pest control. [ABSTRACT FROM AUTHOR]

Many plant secondary substances are feeding deterrents for insects and play a key role in the selection of host plants. The taste sensilla of phytophagous insects contain gustatory receptor neurons sensitive to deterrents but the molecular basis of deterrent chemoreception remains unknown. We investigated the function of Gr180, the most highly expressed bitter gustatory receptor in the maxillary galea of Helicoverpa armigera larvae. Functional analyses using the Xenopus oocyte expression system and two-electrode voltage clamp revealed that the oocytes expressing Gr180 responded to coumarin. Tip recording results showed that the medial sensilla styloconica of the maxilla of fifth instar larvae exhibited electrophysiological responses to coumarin. Two-choice feeding bioassays confirmed that coumarin inhibited larval feeding. A homozygous mutant strain of H. armigera with truncated Gr180 proteins (Gr180−/−) was established using the CRISPR-Cas9 system. The responses of the medial sensilla styloconica in Gr180−/− to coumarin were almost abolished, and the responses to sinigrin and strychnine, were also significantly decreased. Knockout of Gr180 alleviated the feeding deterrent effects of coumarin, sinigrin, and strychnine. Thus, we conclude that Gr180 is a receptor responding to coumarin,and also participates in sensing sinigrin and strychnine. These results enhance our understanding of the gustatory coding mechanisms of phytophagous insects to deterrents. Author summary: Phytophagous insects avoid eating most plants because of the presence of distasteful secondary compounds, which introduces the question of how these compounds are perceived by insects. The cotton bollworm is a very serious crop pest in the Old World, and its caterpillars mainly feed on the reproductive organs of many host plants. Here, we study the function of a gustatory receptor, Gr180, highly expressed in the taste organ of Helicoverpa armigera caterpillars. The oocytes of frogs expressing Gr180 selectively responded to coumarin, a bitter compound widely exist in plants. When this gene was knocked out, the response of the deterrent cells in the medial styloconic sensilla to coumarin disappeared, and the responses to other two deterrents sinigrin and strychnine were also attenuated. However, the inhibitory effect of coumarin on the sucrose cells in the lateral styloconic sensilla was not affected. At the same time, the aversive feeding responses to these compounds were reduced in the knockout caterpillars. These findings are important for revealing the mechanisms of host selection in herbivorous insects and the use of feeding deterrents for pest control. [ABSTRACT FROM AUTHOR]

In the title centrosymmetric dimeric complex of SnIV, [Sn2(CH3)4(C10H7ClN2O3)2(C2H6O)2], each Sn atom has a distorted pentagonal–bipyramidal configuration, being coordinated by three O atoms and one N atom from the pyruvic acid 4-chlorobenzoylhydrazone ligands, one O atom from an ethanol molecule and two axial C atoms from trans methyl groups. [ABSTRACT FROM AUTHOR]

Background: Odorant receptors (ORs) as odorant-gated ion channels play a crucial role in insect olfaction. They are formed by a heteromultimeric complex of the odorant receptor co-receptor (Orco) and a ligand-selective Or. Other types of olfactory receptor proteins, such as ionotropic receptors (IRs) and some gustatory receptors (GRs), are also involved in the olfactory system of insects. Orco as an obligatory subunit of ORs is highly conserved, providing an opportunity to systematically evaluate OR-dependent olfactory responses. Results: Herein, we successfully established a homozygous mutant (Orco−/−) of Helicoverpa armigera, a notorious crop pest, using the CRISPR/Cas9 gene editing technique. We then compared the olfactory response characteristics of wild type (WT) and Orco−/− adults and larvae. Orco−/− males were infertile, while Orco−/− females were fertile. The lifespan of Orco−/− females was longer than that of WT females. The expressions of most Ors, Irs, and other olfaction-related genes in adult antennae of Orco−/− moths were not obviously affected, but some of them were up- or down-regulated. In addition, there was no change in the neuroanatomical phenotype of Orco−/− moths at the level of the antennal lobe (including the macroglomerular complex region of the male). Using EAG and SSR techniques, we discovered that electrophysiological responses of Orco−/− moths to sex pheromone components and many host plant odorants were absent. The upwind flight behaviors toward sex pheromones of Orco−/− males were severely reduced in a wind tunnel experiment. The oviposition selectivity of Orco−/− females to the host plant (green pepper) has completely disappeared, and the chemotaxis toward green pepper was also lost in Orco−/− larvae. Conclusions: Our study indicates that OR-mediated olfaction is essential for pheromone communication, oviposition selection, and larval chemotaxis of H. armigera, suggesting a strategy in which mate searching and host-seeking behaviors of moth pests could be disrupted by inhibiting or silencing Orco expression. [ABSTRACT FROM AUTHOR]

The closely related species Helicoverpa armigera (H. armigera) and Helicoverpa assulta (H. assulta) have different host plant ranges and share two principal components of sex pheromones but with reversed ratios. The antennae are the main olfactory organ of insects and play a crucial role in host plant selection and mate seeking. However, the genetic basis for gene expression divergence in the antennae of the two species is unclear. We performed an allele-specific expression (ASE) analysis in the antennal transcriptomes of the two species and their F1 hybrids, examining the connection between gene expression divergence and phenotypic differences. The results show that the proportion of genes classified as all cis was higher than that of all trans in males and reversed in females. The contribution of regulatory patterns to gene expression divergence in males was less than that in females, which explained the functional differentiation of male and female antennae. Among the five groups of F1 hybrids, the fertile males from the cross of H. armigera female and H. assulta male had the lowest proportion of misexpressed genes, and the inferred regulatory patterns were more accurate. By using this group of F1 hybrids, we discovered that cis-related regulations play a crucial role in gene expression divergence of sex pheromone perception-related proteins. These results are helpful for understanding how specific changes in the gene expression of olfactory-related genes can contribute to rapid evolutionary changes in important olfactory traits in closely related moths. [ABSTRACT FROM AUTHOR]

The ichneumonid wasp, Campoletis chlorideae Uchida, successfully develops in the cotton bollworm Helicoverpa armigera (Hübner), but rarely survives in the beet armyworm Spodoptera exigua (Hübner) due to the encapsulation by host immunity. In this study, we investigated the role of C . chlorideae ichnovirus (CcIV) and eggs in the evasion of the host immune system. Washed eggs of different types, immature, mature, newly laid, or pretreated with protease K, were injected alone or with the calyx fluid containing CcIV into the larvae of H . armigera and S . exigua . In H . armigera , when injected with washed eggs alone, only 9.5% of the mature eggs were encapsulated at 24 h post-injection. This is much lower than that of the immature eggs (100%), mature eggs pretreated with protease K (100%) and newly laid eggs (54.4%). No encapsulation was observed when the washed eggs were co-injected with calyx fluid at 24 h post-injection. Conversely, the eggs in all treatments were encapsulated in S . exigua . Electron microscopic observations of parasitoid eggs showed structural differences between the surfaces of the mature and other kinds of eggs. The injected CcIV decreased the numbers of host hemocytes and suppressed the spreading ability of plasmatocytes and granulocytes in H . armigera , but had little effect on the hemocytes from S . exigua . In conclusion, the C . chlorideae egg provides a passive protection against encapsulation by itself, and CcIV supplies an active protection through disrupting host immune responses. These coordinated protections are host-specific, implying their role in host range determination. [ABSTRACT FROM AUTHOR]

The content of magnetic iron is one of the basic analyses in ultra-low-grade magnetite exploration. In order to accurately determine the content of magnetic iron, quantitative separation of magnetic iron is very important. There is no standard method for the analysis of magnetic iron in iron ore, and the reproducibility of common artificial magnetic separation is poor due to the human factors and the difficulty of guaranteeing effective magnetic field intensity. A new magnetic separation device (electromagnetic separation device) was developed by using 50 mL burette, electromagnet and a three-phase asynchronous motor. Magnetic iron and nonmagnetic iron in ultra-low-grade magnetite was quantitatively separated by optimizing the working conditions. Determination of magnetic iron was proposed by combining the potassium bichromate titrimetric method with a magnetic separation technique. When the current was 2.5 A, the motion frequency of the magnetic tube was 40 times/min, and the time of magnetic separation was 5 min. The relative error was lower than 1.0% for the reference material. For the practical samples, the analysis results completed by the proposed method were in agreement with those determined by artificial magnetic separation with the RSD of the proposed method being lower than 1.0%, which was an improvement over the precision of artificial magnetic separation. For the new method, the magnetic field intensity was effectively controlled, the parameters for the separation of magnetic iron were quantized, and the measurement accuracy was improved for magnetic iron. [ABSTRACT FROM AUTHOR]

In this study of multi-layer tablets, the dissolution of biphasic and zero-order release formulations has been studied primarily using attenuated total reflection- Fourier transform infrared ( ATR- FTIR) spectroscopic imaging as well as UV- Vis detection of dissolved drug in the effluent stream and USP dissolution testing. Bilayer tablets, containing the excipients microcrystalline cellulose ( MCC) and glucose, were used for biphasic release with nicotinamide and buflomedil as model drugs. ATR- FTIR spectroscopic imaging showed the changing component distributions during dissolution. Further experiments studied monolithic and barrier-layered tablets containing hydroxypropyl methylcellulose, MCC and buflomedil dissolving in a USP I apparatus. These data were compared with UV- Vis dissolution profiles obtained online with the ATR flow-through cell. ATR- FTIR imaging data of the biphasic formulations demonstrated that the drug release was affected by excipient ratios and effects such as interference between tablet sections. Tablets placed in the ATR- FTIR flow-through cell exhibited zero-order UV- Vis dissolution profile data at high flow rates, similar to barrier-layered formulations studied using the USP I apparatus. ATR- FTIR spectroscopic imaging provided information regarding the dissolution mechanisms in multi-layer tablets which could assist formulation development. The ability to relate data from USP dissolution tests with that from the ATR- FTIR flow-through cell could help spectroscopic imaging complement dissolution methods used in the industry. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 103:1995-2004, 2014 [ABSTRACT FROM AUTHOR]

Chemosensory proteins (CSPs) are soluble proteins found only in arthropods. Some of them fill the lumen of chemosensilla and are believed to play a role similar to that of odorant-binding proteins in the detection of semiochemicals. Other members of the CSP family have been reported to perform different functions, from delivery of pheromones to development. This report is focused on a member (CSP4) of the family that is highly and almost exclusively present in the proboscis of two sibling noctuid species, Helicoverpa armigera and H. assulta. We expressed the protein in bacteria and measured binding to terpenoids and related compounds. Using specific antibodies, we found that when the moths suck on a sugar solution, CSP4 is partly extruded from the proboscis. A solution of protein can also fill a hydrophobic tube of same length and diameter as the proboscis by capillary action. On this basis, we suggest that CSP4 acts as a wetting agent to reduce the surface tension of aqueous solutions and consequently the pressure involved in sucking. [ABSTRACT FROM AUTHOR]

The relative proportions of components in a pheromone blend play a major role in sexual recognition in moths. Two sympatric species, Helicoverpa armigera and Helicoverpa assulta, use (Z)-11-hexadecenal (Z11–16: Ald) and (Z)-9-hexadecenal (Z9–16: Ald) as essential sex pheromone components but in very different ratios, 97∶3 and 7∶93 respectively. Using wind tunnel tests, single sensillum recording and in vivo calcium imaging, we comparatively studied behavioral responses and physiological activities at the level of antennal sensilla and antennal lobe (AL) in males of the two species to blends of the two pheromone components in different ratios (100∶0, 97∶3, 50∶50, 7∶93, 0∶100). Z11–16: Ald and Z9–16: Ald were recognized by two populations of olfactory sensory neurons (OSNs) in different trichoid sensilla on antennae of both species. The ratios of OSNs responding to Z11–16:Ald and Z9–16:Ald OSNs were 100∶28.9 and 21.9∶100 in H. armigera and H. assulta, respectively. The Z11–16:Ald OSNs in H. armigera exhibited higher sensitivity and efficacy than those in H. assulta, while the Z9–16:Ald OSNs in H. armigera had the same sensitivity but lower efficacy than those in H. assulta. At the dosage of 10 µg, Z11–16: Ald and Z9–16: Ald evoked calcium activity in 8.5% and 3.0% of the AL surface in H. armigera, while 5.4% and 8.6% of AL in H. assulta, respectively. The calcium activities in the AL reflected the peripheral input signals of the binary pheromone mixtures and correlated with the behavioral output. These results demonstrate that the binary pheromone blends were precisely coded by the firing frequency of individual OSNs tuned to Z11–16: Ald or Z9–16: Ald, as well as their population sizes. Such information was then accurately reported to ALs of H. armigera and H. assulta, eventually producing different behaviors. [ABSTRACT FROM AUTHOR]

Some of the components found in herbs may be inhibitors or inducers of cytochrome P450 enzymes, which may therefore result in undesired herb-drug interactions. As a component extracted from Radix Scutellariae, the direct effect of baicalin on cytochrome P450 has not been investigated sufficiently. In this study, we investigated concentration-dependent inhibitory effect of baicalin on the plasma protein binding and metabolism of chlorzoxazone (CZN), a model CYP2E1 probe substrate, in rats in vitro and in vivo. Animal experiment was a randomized, three-period crossover design. Significant changes in pharmacokinetic parameters of CZN such as Cmax, t1/2 and Vd were observed after treatment with baicalin in vivo (P<0.05). Cmax decreased by 25% and 33%, whereas t1/2 increased by 34% and 53%, Vd increased by 37% and 50% in 225 mg/kg and 450 mg/kg baicalin-treated rats, respectively. The AUC and CL of CZN were not affected (P.0.05). Correlation analysis showed that the changes in CZN concentrations and baicalin concentrations were in good correlation (r>0.99). In vitro experiments, baicalin decreased the formation of 6-OH-chlorzoxazone in a concentration-dependent manner and exhibited a competitive inhibition in rat liver microsomes, with a Ki value of 145.8 mM. The values of Cmax/Ki were 20 and 39 after treatment with baicalin (225 and 450 mg/kg), respectively. Protein binding experiments in vivo showed that the plasma free-fraction (fu) of CZN increased 2.6-fold immediately after baicalin treatment (450 mg/kg) and in vitro showed that baicalin (125-2500 mg/L) increased the unbound CZN from 1.63% to 3.58%. The results indicate that pharmacokinetic changes in CZN are induced by inhibitory effect of baicalin on the plasma protein binding of CZN and CYP2E1 activity. [ABSTRACT FROM AUTHOR]

The interleukin (IL)-4 and IL-4R genes are linked to allergic diseases and atopy and elevated serum IgE levels closely follow the presentation of penicillin allergy. We have evaluated the association between specific immunoglobulin (Ig)E, polymorphisms of IL-4Rα, and penicillin allergy. Eight types of specific IgE to penicillins were detected with the radioallergosorbent test (RAST) in 242 patients with penicillin allergy and 220 control subjects. Polymorphisms of IL-4Rα Q576R and IL-4Rα I75V were genotyped by PCR–restriction fragment length polymorphism (RFLP). The polymorphisms and haplotype of IL-4Rα Q576R and I75V were analyzed. For IL-4Rα Q576R, the frequency of the AA genotype was higher in patients with penicillin allergy (76 vs. 64%, P = 0.005). The A allele occurred more frequently in the penicillin-allergic patients than in the controls (87 vs. 80%, P = 0.003). For IL-4Rα I75V, the AA genotype was more likely to be detected in the urticaria group than in the control group (32 vs. 19%, P = 0.049). Haplotype analysis revealed that Q576/I75 was more frequently found in patients with penicillin allergy than in controls (42 vs. 35%, P = 0.037). The IL4Rα Q576 allele is related to penicillin allergy, and the IL-4Rα I75 allele is associated with the symptom of urticaria. The Q576/I75 haplotype may be related with an allergy to penicillin. [ABSTRACT FROM AUTHOR]

Ribosomal protein S13 gene has been cloned and analyzed in many organisms, but there are few documents relating to insects. In this communication, the full-length cDNA sequence of ribosomal protein S13 gene in the diamondback moth,Plutella xylostella(Lepidoptera: Plutellidae), was determined by using PCR amplification technique. The features of the ribosomal protein S13 gene sequence were analyzed and the deduced amino acids sequence was compared with those from other insects. The results of multi-alignment of the amino acid sequences between the diamondback moth and other insect species revealed that this gene sequence is highly conserved in insects. Based on maximum likelihood method, a phylogenetic tree was constructed from 10 different species using PHYLIP software. It showed that nematode is one separate lineage and the five insect species belong to another lineage, whereas those species higher than insects form the third one. The pattern of this phylogenetic tree evidently represented the evolution of different species. [ABSTRACT FROM AUTHOR]

Glucosinolates are token stimuli in host selection of many crucifer specialist insects, but the underlying molecular basis for host selection in these insects remains enigmatic. Using a combination of behavioral, electrophysiological, and molecular methods, we investigate glucosinolate receptors in the cabbage butterfly Pieris rapae. Sinigrin, as a potent feeding stimulant, elicited activity in larval maxillary lateral sensilla styloconica, as well as in adult medial tarsal sensilla. Two P. rapae gustatory receptor genes PrapGr28 and PrapGr15 were identified with high expression in female tarsi, and the subsequent functional analyses showed that Xenopus oocytes only expressing PrapGr28 had specific responses to sinigrin; when ectopically expressed in Drosophila sugar sensing neurons, PrapGr28 conferred sinigrin sensitivity to these neurons. RNA interference experiments further showed that knockdown of PrapGr28 reduced the sensitivity of adult medial tarsal sensilla to sinigrin. Taken together, we conclude that PrapGr28 is a gustatory receptor tuned to sinigrin in P. rapae, which paves the way for revealing the molecular basis of the relationships between crucifer plants and their specialist insects. Author summary: Preference of crucifer specialist insects to glucosinolates is well known in the field of insect-plant interactions, but its molecular basis is unclear. This study uses an integrative approach to investigate the molecular basis of glucosinolate detection by gustatory receptor neurons in the larval mouthparts and adult forelegs of the cabbage butterfly Pieris rapae, and finally reveal that PrapGr28 is a bitter receptor tuned to sinigrin. The current work takes a significant step towards identifying gustatory receptors tuned to glucosinolates, crucial recognition signals in crucifer host plants, providing insights into co-evolution of herbivorous insects and their host plants. [ABSTRACT FROM AUTHOR]

The aim was to evaluate the efficacy, safety and completion rate of 3-month, once-weekly rifapentine and isoniazid for tuberculosis (TB) prevention among Chinese silicosis patients. Male silicosis patients without human immunodeficiency virus infection, aged 18 years to 65 years, with or without latent TB infection, were randomized 1:1 to receive rifapentine/isoniazid under direct observation (3RPT/INH group) or were untreated (observation group). Active TB incidence was compared between the two groups with 37 months of follow-up. Safety profile and complete rates were evaluated. A total of 1227 adults with silicosis were screened; 513 eligible participants were enrolled and assigned to 3RPT/INH (n = 254) vs. observation (n = 259). Twenty-eight participants were diagnosed with active TB, and 9 and 19 in the 3RPT/INH group and observation groups, respectively. In the intention-to-treat analysis, the cumulative active TB rate was 3.5% (9/254) in the 3RPT/INH group and 7.3% (19/259) in the observation group (log rank p 0.055). On per protocol analysis, the cumulative active TB rates were 0.7% (1/139) and 7.3% (19/259), respectively (log rank p 0.01). Owing to an unexpected high frequency of adverse events (70.4%) and Grade 3 or 4 AEs (7.9%), the completion rate of the 3RPT/INH regimen was 54.7% (139/254). Twenty-six (10.8%) participants had flu-like systemic drug reactions; five (2.1%) experienced hepatotoxicity. Weekly rifapentine/isoniazid prophylaxis prevented active TB among Chinese people with silicosis when taken, irrespective of LTBI screening; efficacy was reduced by lack of compliance. The regimen must be used with caution because of the high rates of adverse effects. ClinicalTrials.gov number: NCT02430259 [ABSTRACT FROM AUTHOR]

Feeding and oviposition deterrents help phytophagous insects to identify host plants. The taste organs of phytophagous insects contain bitter gustatory receptors (GRs). To explore their function, the GRs in Plutella xylostella were analyzed. Through RNA sequencing and qPCR, we detected abundant PxylGr34 transcripts in the larval head and adult antennae. Functional analyses using the Xenopus oocyte expression system and 24 diverse phytochemicals showed that PxylGr34 is tuned to the canonical plant hormones brassinolide (BL) and 24-epibrassinolide (EBL). Electrophysiological analyses revealed that the medial sensilla styloconica of 4th instar larvae are responsive to BL and EBL. Dual-choice bioassays demonstrated that BL inhibits larval feeding and female oviposition. Knock-down of PxylGr34 by RNAi attenuates the taste responses to BL, and abolishes BL-induced feeding inhibition. These results increase our understanding of how herbivorous insects detect compounds that deter feeding and oviposition, and may be useful for designing plant hormone-based pest management strategies. [ABSTRACT FROM AUTHOR]

Recent studies have shown that microcystin-LR (MC-LR) is one of the principal factors that cause liver cancer. Previously we have found that Aristaless-like Homeobox 4 (ALX4) was differentially expressed in MC-LR-induced malignant transformed L02 cells. However, the expression regulation, role and molecular mechanism of ALX4 during the process of liver cancer induced by MC-LR are still unclear. The expression of ALX4 was detected by quantitative reverse-transcription PCR and Western blot in MC-LR induced malignantly transformed cell and rat models. Methylation status of ALX4 promoter region was evaluated by methylation-specific PCR and bisulfite genomic sequencing. The anti-tumor effects of ALX4 on MC-LR induced liver cancer were identified in vitro and in vivo. ALX4 expression was progressively down-regulated in MC-LR-induced malignantly transformed L02 cells and the MC-LR exposed rat models. ALX4 promoter regions were highly methylated in malignantly transformed cells, while treatment with demethylation agent 5-aza-dC significantly increased ALX4 expression. Functional studies showed that overexpression of ALX4 inhibits cell proliferation, migration, invasion and metastasis in vitro and in vivo , blocks the G1/S phase and promotes the apoptosis. Conversely, knockdown of ALX4 promotes cell proliferation, migration and invasion. Mechanism study found that ALX4 exerts its antitumor function through the P53 pathway, C-MYC and MMP9. More importantly, ALX4 expression level showed a negative relation with serum MC-LR levels in patients with hepatocellular carcinoma. Our results suggested that ALX4 was inactivated by DNA methylation and played a tumor suppressor function through the P53 pathway in MC-LR induced liver cancer. Unlabelled Image • ALX4 expression was progressively down-regulated in MC-LR-induced hepatocarcinogenesis. • DNA methylation regulated ALX4 gene expression during MC-LR induced hepatocarcinogenesis. • ALX4 inhibited growth, migration and invasion of MC-LR induced malignantly transformed L02 cells in vitro and in vivo. • ALX4 plays a tumor suppressing role in MC-LR-induced liver cancer through the P53 pathway, C-MYC and MMP9. • ALX4 expression level showed a negative relation with serum MC-LR levels in patients with hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]

Aims: To evaluate the feasibility of assessing regional myocardial perfusion using real-time myocardial contrast echocardiography (MCE) at rest for detecting coronary microcirculation abnormalities in methamphetamine abusers.Material and methods: Twenty-two male methamphetamine abusers (11 without chest pain, 11 with chest pain), free of ascertained coronary artery disease, were enrolled in this study. A control group of 22 age-matched male healthy participants was studied for comparison. Standard 2D, flow and tissue Doppler echo with measurements of cardiac morphologic and functional indicators,MCE with measurements of regional myocardial perfusion were performed, respectively.Results: Compared to healthy participants, methamphetamine abusers had higher blood pressure, greater left ventricular mass index and more impaired diastolic function, with preserved cardiac sizes and systolic function. Methamphetamine abusers with chest pain had a faster heart rate than those without chest pain and healthy participants. MCE in methamphetamine abusers, especially with chest pain, had significant longer contrast agent arrival times, less functional capillary blood volumes, slower microvascular flow velocities and less myocardial perfusion than healthy participants (p<0.05). Moreover, along with the increases of dosage and duration of use (from group A to group C, group A: 1-2 g/day, <2 years; group B: 2-3 g/day, 2-5 years; group C: >3 g/day, >5 years) the reductions in the myocardial perfusion indices were more significant (p<0.01). The cutoff value with 5.1 dB2/s of the myocardial perfusion at the left ventricular apex had a sensitivity of 87.5%, specificity of 75.2% and accuracy of 81.9% for differentiating methamphetamine abusers from normal subjects.Conclusions: Real-time MCE can effectively detect coronary microcirculation abnormalities in methamphetamine abusers at rest and myocardial perfusion is significantly reduced in methamphetamine abusers. This finding may be involved in the occurrence and development of cardiac damage. [ABSTRACT FROM AUTHOR]

The phytochemical investigation of the EtOAc extract from the aerial parts of Isodon eriocalyx afforded seventeen diterpenoids, including eight undescribed compounds. Eriocalyxins H−L have unique structural characteristics featuring a 5- epi - ent -kaurane diterpenoid scaffold with eriocalyxins H–K also possess an unusual 6,11-epoxyspiro-lactone ring while eriocalyxin L, a 1,7:3,20-diepoxy- ent kaurene, features an 1,7-oxygen linkage. The structures of these compounds were elucidated by spectroscopic data interpretation, and the absolute configurations of eriocalyxins H, I, L, and M were confirmed by single-crystal X-ray diffraction. The isolates were screened for their inhibitory activities against VCAM-1 and ICAM-1 at 5 μM. While eriocalyxin O, coetsoidin A and laxiflorin P were found to significantly inhibit both VCAM-1 and ICAM-1, 8 (17),13- ent -labdadien-15 → 16-lactone-19-oic acid displayed evidently inhibitory effect against ICAM-1. Eight undescribed diterpenoids and nine known ones were isolated from the EtOAc extract of the aerial parts of I. eriocalyx. Their inhibitory activities against VCAM-1 and ICAM-1 were evaluated. [Display omitted] • Eight undescribed ent -kaurane diterpenoids were isolated from Isodon eriocalyx. • Eriocalyxins H−L represent the first examples of 5- epi - ent -kaurane diterpenoids. • Eriocalyxins H–K are the first ent -kauranoids with a 6,11-epoxyspiro-lactone scaffold. • Eriocalyxin L is the first 1,7:3,20-diepoxy- ent -kaurane diterpenoid. • Four kauranoids displayed significant activities against atherosclerosis. [ABSTRACT FROM AUTHOR]

Determining drug-metabolizing enzyme activities on an individual basis is an important component of personalized medicine, and cytochrome P450 enzymes (CYPs) play a principal role in hepatic drug metabolism. Herein, a simple method for predicting the major CYP-mediated drug clearance in vitro and in vivo is presented. Ten CYP-mediated drug metabolic activities in human liver microsomes (HLMs) from 105 normal liver samples were determined. The descriptive models for predicting the activities of these CYPs in HLMs were developed solely on the basis of the measured activities of a smaller number of more readily assayed CYPs. The descriptive models then were combined with the Conventional Bias Corrected in vitro–in vivo extrapolation method to extrapolate drug clearance in vivo. The Vmax, Km, and CL int of six CYPs (CYP2A6, 2C8, 2D6, 2E1, and 3A4/5) could be predicted by measuring the activities of four CYPs (CYP1A2, 2B6, 2C9, and 2C19) in HLMs. Based on the predicted CL int, the values of CYP2A6-, 2C8-, 2D6-, 2E1-, and 3A4/5-mediated drug clearance in vivo were extrapolated and found that the values for all five drugs were close to the observed clearance in vivo. The percentage of extrapolated values of clearance in vivo which fell within 2-fold of the observed clearance ranged from 75.2% to 98.1%. These findings suggest that measuring the activity of CYP1A2, 2B6, 2C9, and 2C19 allowed us to accurately predict CYP2A6-, 2C8-, 2D6-, 2E1-, and 3A4/5-mediated activities in vitro and in vivo and may possibly be helpful for the assessment of an individual’s drug metabolic profile. [ABSTRACT FROM AUTHOR]

CQMUH-011, a novel adamantane sulfonamide compound, was shown to suppress macrophage activation and proliferation in our previous study. However, it is unknown whether CQMUH-011 has anti-inflammatory and hepatoprotective properties. In this study, we investigated the potential effects and mechanisms of CQMUH-011 on lipopolysaccharide (LPS)-induced RAW264.7 cell activation in vitro and LPS- and D-galactosamine (D-GalN)-induced fulminant hepatic failure (FHF) in vivo . The results showed that in RAW264.7 cells challenged by LPS, CQMUH-011 inhibited cell proliferation and induced cell cycle arrest and apoptosis. Furthermore, CQMUH-011 reduced tumor necrosis factor (TNF)-α and interleukin (IL)-1β production and down-regulated the overexpression of toll-like receptor 4 (TLR4) and nuclear factor (NF)-κB induced by LPS in RAW264.7 cells. In vivo , CQMUH-011 reduced serum levels of aspartic aminotransferase and alanine transaminase and improved the mortality and hepatic pathological damage induced by LPS/D-GalN in mice. Moreover, CQMUH-011 significantly inhibited the serum levels of proinflammatory mediators, including TNF-α, IL-6, IL-1β, nitric oxide (NO), and prostaglandin E2 (PGE 2 ), and down-regulated the protein expression of TLR4, p38 mitogen-activated protein kinases, NF-κB, NF-κB inhibitor α (IκBα), IκB kinase β (IKKβ), cyclooxygenase-2 (COX-2) and inducible NO synthases (iNOS) induced by LPS/D-GalN in mice. In conclusion, these results demonstrated that CQMUH-011 has a notable anti-inflammatory effect and protects mice from LPS/D-GalN-induced FHF and that the molecular mechanisms might be related to the inhibition of the TLR4/NF-κB signaling pathway activation, the subsequent decrease in proinflammatory mediator production, and the inhibition of macrophage activation. [ABSTRACT FROM AUTHOR]

Adult moths possess an organ in their labial palps, the labial-palp pit organ, which is specialized for sensing carbon dioxide (CO 2 ). They use CO 2 as a cue to detect healthy plants and find food or lay eggs on them. The molecular bases of the CO 2 receptor in Drosophila melanogaster and Aedes aegypti have been reported, but the molecular mechanisms of the CO 2 receptor in Lepidoptera remains elusive. In this study, we first re-examined three putative Helicoverpa armigera CO 2 gustatory receptor genes ( HarmGr1 , HarmGr2 , and HarmGr3 ), and then analyzed expression patterns of them. RT-PCR results verified they were predominantly expressed in the labial palps of H. armigera. Thus, we used in situ hybridization to localize the expression of three genes in the labial palps. We found that all three genes were co-expressed in the same cells of the labial palps. Next, we employed the Xenopus laevis oocyte expression system and the two-electrode voltage-clamp recording to study the function of the three genes. Results showed that only oocytes co-expressing HarmGr1 and HarmGr3 or co-expressing HarmGr1, HarmGr2 and HarmGr3 gave robust responses to NaHCO 3 . Finally, we confirmed that the sensory cells in labial palps of both females and males show dose dependent responses to CO 2 stimuli by using single sensillum recording. Our work uncovers that HarmGr1 and HarmGr3 are indispensable and sufficient for CO 2 sensing in labial palps of H. armigera . [ABSTRACT FROM AUTHOR]

Insect gustatory systems play important roles in food selection and feeding behaviors. In spite of the enormous progress in understanding gustation in Drosophila , for other insects one of the key elements in gustatory signaling, the gustatory receptor (GR), is still elusive. In this study, we report that fructose elicits behavioral and physiological responses in Helicoverpa armigera (Harm) to fructose and identify the gustatory receptor for this sugar. Using the proboscis extension reflex (PER) assays we found that females respond to fructose following stimulation of the distal part of the antenna, where we have identified contact chemosensilla tuned to fructose in tip recording experiments. We isolated three full-length cDNAs encoding candidate HarmGRs based on comparison with orthologous GR sequences in Heliothis virescens and functionally characterized the responses of HarmGR4 to 15 chemicals when this receptor was expressed in Xenopus oocytes with two-electrode voltage-clamp recording. Among the tastants tested, the oocytes dose-dependently responded only to d -fructose (EC 50 = 0.045 M). By combining behavioral, electrophysiological and molecular approaches, these results provide basic knowledge for further research on the molecular mechanisms of gustatory reception. [ABSTRACT FROM AUTHOR]

Tap water (unfiltered), filtered tap water and processed bottled water (purified water, artificial mineralized water, or natural water) are now the five most widely consumed types of drinking water in China. However, the constituents (organic chemicals and inorganic ingredients) of the five waters differ, which may cause them to have different long-term health effects on those who drink them, especially sensitive children. In order to determine which type of water among the five waters is the most beneficial regarding reproductive outcomes and the developmental behaviors of offspring, two generations of Sprague–Dawley rats were given these five waters separately, and their reproductive outcomes and the developmental behaviors of their offspring were observed and compared. The results showed that the unfiltered tap water group had the lowest values for the maternal gestation index (MGI) and offspring's learning and memory abilities (OLMA); the lowest offspring survival rate was found in the purified water group; and the highest OLMA were found in the filtered tap water group. Thus, the best reproductive and offspring early developmental outcomes were found in the group that drank filtered tap water, which had the lowest levels of pollutants and the richest minerals. Therefore, thoroughly removing toxic contaminants and retaining the beneficial minerals in drinking water may be important for both pregnant women and children, and the best way to treat water may be with granular activated carbon and ion exchange by copper zinc alloy. [ABSTRACT FROM AUTHOR]

Angiogenesis plays an important role in myocardial infarction. Apelin and its natural receptor (angiotensin II receptor-like 1, AGTRL-1 or APLNR) induce sprouting of endothelial cells in an autocrine or paracrine manner. The aim of this study is to investigate whether apelin can improve the cardiac function after myocardial infarction by increasing angiogenesis in infarcted myocardium. Left ventricular end-diastolic pressure (LVEDP), left ventricular end systolic pressure (LVESP), left ventricular developed pressure (LVDP), maximal left ventricular pressure development (±LVd p/d tmax), infarct size, and angiogenesis were evaluated to analyze the cardioprotective effects of apelin on ischemic myocardium. Assays of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, 5-bromo-2′-deoxyuridine incorporation, wound healing, transwells, and tube formation were used to detect the effects of apelin on proliferation, migration, and chemotaxis of cardiac microvascular endothelial cells. Fluorescein isothiocyanate-labeled bovine serum albumin penetrating through monolayered cardiac microvascular endothelial cells was measured to evaluate the effects of apelin on permeability of microvascular endothelial cells. In vivo results showed that apelin increased ±LV d p/d tmax and LVESP values, decreased LVEDP values (all p < 0.05), and promoted angiogenesis in rat heart after ligation of the left anterior descending coronary artery. In vitro results showed that apelin dose-dependently enhanced proliferation, migration, chemotaxis, and tube formation, but not permeability of cardiac microvascular endothelial cells. Apelin also increased the expression of vascular endothelial growth factor receptors-2 (VEGFR2) and the endothelium-specific receptor tyrosine kinase (Tie-2) in cardiac microvascular endothelial cells. These results indicated that apelin played a protective role in myocardial infarction through promoting angiogenesis and decreasing permeability of microvascular endothelial cells via upregulating the expression of VEGFR2 and Tie-2 in cardiac microvascular endothelial cells. [ABSTRACT FROM AUTHOR]

Abstract: The olfactory system of moth species in subfamily Heliothinae is an attractive model to study the evolution of the pheromone reception because they show distinct differentiation in sex pheromone components or ratios that activate pheromone receptors (PRs). However, functional assessment of PRs in closely related species remains largely untried. Here we present a special cloning strategy to isolate full-length cDNAs encoding candidate odorant receptors (ORs) from Helicoverpa armigera (Harm) and Helicoverpa assulta (Hass) on the basis of Heliothis virescens ORs, and investigate the functional properties of PRs to determine how the evolution of moth PRs contribute to intraspecific mating choice and speciation extension. We cloned 11 OR orthologs from H. armigera and 10 from H. assulta. We functionally characterized the responses of PRs of both species to seven pheromone compounds using the heterologous expression system of Xenopus ooctyes. HassOR13 was found to be highly tuned to the sex pheromone component Z11-16:Ald, and unexpectedly, both HarmOR14b and HassOR16 were specific for Z9-14:Ald. However, HarmOR6 and HassOR6 showed much higher specificity to Z9-16:OH than to Z9-16:Ald or Z9-14:Ald. HarmOR11, HarmOR14a, HassOR11 and HassOR14b failed to respond to the tested chemicals. Based on our results and previous research, we can show that some PR orthologs from H. armigera, H. assulta and H. virescens such as OR13s have similar ligand selectivity, but others have different ligand specificity. The combined PR function and sex pheromone component analysis suggests that the evolution of PRs can meet species-specific demands. [Copyright &y& Elsevier]

Baicalin, a flavonoid compound isolated from Scutellaria baicalensis, has been shown to possess antiinflammatory, antiviral, antitumour, and immune regulatory properties. The present study evaluated the potential herb-drug interaction between baicalin and midazolam in rats. Coadministration of a single dose of baicalin (0.225, 0.45, and 0.90 g/kg, i.v.) with midazolam (10 mg/kg, i.v.) in rats resulted in a dose-dependent decrease in clearance (CL) from 25% (P < 0.05) to 34% (P < 0.001) with an increase in AUC0-∞ from 47% (P < 0.05) to 53% (P < 0.01). Pretreatment of baicalin (0.90g/kg, i.V., once daily for 7 days) also reduced midazolam CL by 43% (P < 0.001), with an increase in AUC0-∞ by 87% (P < 0.01). Multiple doses of baicalin decreased the expression of hepatic CYP3A2 by approximately 58% (P < 0.01) and reduced midazolam 1 -hydroxylation by 23% (P < 0.001) and 4'-hydroxylation by 21% (P < 0.01) in the liver. In addition, baicalin competitively inhibited midazolam metabolism in rat liver microsomes in a concentration-dependent manner. Our data demonstrated that baicalin induced changes in the pharmacokinetics of midazolam in rats, which might be due to its inhibition of the hydroxylation activity and expression of CYP3A in the liver. [ABSTRACT FROM AUTHOR]

Abstract: Short, nonlethal ischemic episodes administered to hearts directly after ischemic events (ischemic postconditioning, IPost) have an advantage over ischemic preconditioning (IPC). The endogenous cytochrome P450 2J3/11,12-epoxyeicosatrienoic acid (CYP2J3/11,12-EET) is upregulated by IPost, but not IPC, in the rat heart. The CYP epoxygenase inhibitor N-methylsulphonyl-6-(2-propargyloxyphenyl) hexanamide (MS-PPOH) reduces the cardioprotective effects of IPost, but not IPC. We proposed that upregulation of CYP2J3/11,12-EET during IPost induces cardioprotection by inhibiting cardiomyocyte apoptosis and that multiple apoptotic signals, including changes in mitochondrial membrane potential (MMP) and mitochondrial permeability transition pore (mPTP) opening, mitochondrial cytochrome c leakage, caspase-3 levels, and levels of protective kinases such as Bcl-2 and Bax, are involved in the process. Neonatal rat cardiomyocytes underwent 3-h hypoxia followed by 2-, 5-, or 6-h reoxygenation (H/R) or three cycles of 5-min reoxygenation followed by 5-min hypoxia before 90-min reoxygenation (HPost); or were transfected with pcDNA3.1-CYP2J3 for 48h before H/R; or were treated with MS-PPOH for 10min before HPost. For HPost alone, pcDNA3.1-CYP2J3 transfection attenuated cardiomyocyte apoptosis to 68.4% (p <0.05) of that with H/R. pcDNA3.1-CYP2J3 transfection significantly decreased MMP and inhibited mPTP opening induced by H/R, reduced mitochondrial cytochrome c leakage, cleaved caspase-3 protein expression, and increased the ratio of Bcl-2 to Bax expression. MS-PPOH abolished this effect. Therefore, upregulation of CYP2J3/11,12-EET during HPost is involved in cardioprotection by inhibiting apoptosis via a caspase-dependent pathway, and the apoptosis-suppressive effect may have important clinical implications during HPost. [Copyright &y& Elsevier]

1. Cytochrome P450 (CYP) epoxygenases and their arachidonic acid metabolites play a protective role against ischaemia-reperfusion injury. In the present study, we investigated whether endogenous CYP2J3/epoxyeicosatrienoic acid (EET) mediates the cardioprotective effects of ischaemic preconditioning (IPC) and ischaemic post-conditioning (IPost). 2. Male Wistar rats were subjected to two cycles of IPC, consisting of 5 min ischaemia and 5 min reperfusion, followed by 45 min occlusion and 2 h reperfusion; IPost consisted of three cycles of 30 s reperfusion and 30 s re-occlusion at the onset of reperfusion. The selective CYP epoxygenase inhibitor N-methylsulphonyl-6-(2-propargyloxyphenyl)hexanamide (MS-PPOH; 3 mg/kg) was administered 10 min before ischaemia or during ischaemia 10 min before reperfusion started. Cardiac function was measured continuously with a angiocatheter connected to a fluid-filled pressure transducer and myocardial infarct size was assessed by triphenyl tetrazolium chloride staining at the end of the experiment. 3. Subjecting rats to IPC and IPost similarly improved cardiac function and reduced myocardial infarct size. Interestingly, IPost, but not IPC, significantly increased CYP2J3 mRNA (1.75 ± 0.22 vs 1.0; P < 0.05) and protein (1.62 ± 0.22 vs 1.0; P < 0.05), as well as 11,12-EET synthesis compared to I/R (6.2 ± 0.2 vs 2.9 ± 0.2 ng/mg wet weight, respectively; P < 0.01). Administration of MS-PPOH before ischaemia significantly decreased 11,12-EET synthesis in both IPC and IPost compared with I/R rats (2.1 ± 0.2, 3.2 ± 0.3 and 2.9 ± 0.2 ng/mg wet weight, respectively; P < 0.01), but decreased the cardioprotective effects, as evidenced by cardiac function and myocardial infarct size, of IPost only. 4. These data indicate that endogenous activation of CYP2J3/EET may be an essential trigger leading to the protective effects of IPost, but not IPC, in the rat heart. [ABSTRACT FROM AUTHOR]

Abstract: Apelin, the endogenous ligand of the G protein-coupled APJ receptor, is a peptide mediator with emerging regulatory actions in the heart. We aimed to determine whether the endogenous apelin/APJ system is an intrinsic protective pathway in ischemic/reperfusion injury. A Langendorff model of perfused isolated rat hearts and primary cultured myocardial cells from neonatal rats were used. Cardiac function was monitored and apelin/APJ expression was determined by real-time PCR and Western blot analysis. In rats under I/R, cardiac function was significantly decreased as compared with controls, and APJ was over-expressed at both the mRNA and protein levels (by 7-fold and 35%, respectively, both p <0.01). However, pre-administration of apelin (30pmol/L) greatly ameliorated the reduced heart function. To gain mechanistic insight into the cardio-protective effects of apelin/APJ, cultured cardiomyocytes were treated with apelin (30pmol/L), and those under hypoxia/re-oxygenation showed H/R-induced apoptosis and up-regulated apelin/APJ mRNA expression by 6-fold and 7-fold, respectively (both p <0.01). And lactate dehydrogenase leakage was greatly increased as well. Meanwhile, apoptosis, the generation of reactive oxygen species and malonaldehyde content as well as lactate dehydrogenase leakage were inhibited by apelin. Furthermore, apelin enhanced superoxide dismutase activity and phosphorylation of extracellular signal-regulated kinase 1/2 and Akt after hypoxia/re-oxygenation. In conclusion, apelin/APJ has protective effects in ischemic heart disease and might constitute an important therapy target. [Copyright &y& Elsevier]