A mitochondrial EglN1-AMPKα axis drives breast cancer progression by enhancing metabolic adaptation to hypoxic stress.

Mitochondria play essential roles in cancer cell adaptation to hypoxia, but the underlying mechanisms remain elusive. Through mitochondrial proteomic profiling, we here find that the prolyl hydroxylase EglN1 (PHD2) accumulates on mitochondria under hypoxia. EglN1 substrate-binding region in the ß2ß3 loop is responsible for its mitochondrial translocation and contributes to breast tumor growth. Furthermore, we identify AMP-activated protein kinase alpha (AMPKα) as an EglN1 substrate on mitochondria. The EglN1-AMPKα interaction is essential for their mutual mitochondrial translocation. After EglN1 prolyl-hydroxylates AMPKα under normoxia, they rapidly dissociate following prolyl-hydroxylation, leading to their immediate release from mitochondria. In contrast, hypoxia results in constant EglN1-AMPKα interaction and their accumulation on mitochondria, leading to the formation of a Ca2+ /calmodulin-dependent protein kinase 2 (CaMKK2)-EglN1-AMPKα complex to activate AMPKα phosphorylation, ensuring metabolic homeostasis and breast tumor growth. Our findings identify EglN1 as an oxygen-sensitive metabolic checkpoint signaling hypoxic stress to mitochondria through its ß2ß3 loop region, suggesting a potential therapeutic target for breast cancer.

Insecticide resistance in the Cydia pomonella (L): Global status, mechanisms, and research directions.

The codling moth, Cydia pomonella (Lepidoptera: Tortricidae) is a major pest of pome fruit and walnuts worldwide. Although environmentally compatible integrated control strategies, such as mating disruption, attract-kill strategy, and sterile insect technique have been conducted for management of this notorious pest, effects to control of codling moth have mainly relied on insecticides. In consequence, different levels of insecticide resistance towards organophosphates, neonicotinoids, hydrazines, benzoylureas, pyrethroids, diamides, spinosyns, avermectins, JH mimics, carbamates, oxadiazines and C. pomonella granulovirus (CpGVs) have developed in codling moth in different countries and areas. Both metabolic and target-site mechanisms conferring resistance have been revealed in the codling moth. In this review, we summarize the current global status of insecticide resistance, the biochemical and molecular mechanisms involved, and the implications for resistance management.

Soybean glycinin impaired immune function and caused inflammation associated with PKC-ζ/NF-κb and mTORC1 signaling in the intestine of juvenile grass carp (Ctenopharyngodon idella).

Glycinin is a major protein and antinutritional factor of soybean. However, how dietary glycinin affect intestinal immune function of fish were largely unknown. In this study, we used juvenile grass carp as a model to investigate the impacts of glycinin on intestinal immune function of fish and involved mechanisms. We set three treatments including control, glycinin and glycinin + glutamine in this trial. For immune components, results revealed that compared with control group, glycinin group had lower acid phosphatase activities in the foregut, midgut and hindgut, lower C3 and C4 content, and lower mRNA abundances of IgM, IgZ, hepcidin, LEAP-2A, LEAP-2B and ß-defensin-1 in the midgut and hindgut rather than foregut of grass carp. For pro-inflammatory cytokines and relevant signaling, glycinin elevated mRNA abundances of IL-1ß, IL-8, IL-12p35, IL-12p40 and IL-17D in the midgut and IL-1ß, IFN-γ2, IL-6, IL-8, IL-12p35, IL-12p40 and IL-17D in the hindgut, and increased protein abundances of PKC-ζ and nuclear NF-κB p65 in the midgut and hindgut in comparison to control. For anti-inflammatory cytokines and relevant signaling, glycinin reduced mRNA abundances of TGF-ß1, TGF-ß2, IL-4/13B (rather than IL-4/13A), IL-10 and IL-11 in the midgut and hindgut, and reduced p-mTOR (Ser 2448), p-S6K1 (Thr 389) and p-4EBP1 (Thr 37/46) protein abundances in the midgut and hindgut rather than foregut. Co-administration of glutamine with glycinin could partially enhance intestinal function and reduce intestinal inflammation compared with glycinin treatment. Concluded, glycinin decreased intestinal immune components and caused intestinal inflammation associated with PKC-ζ/NF-κB and mTORC1 signaling.

Soybean glycinin decreased growth performance, impaired intestinal health, and amino acid absorption capacity of juvenile grass carp (Ctenopharyngodon idella).

The present study evaluated the influence of dietary soybean glycinin on growth performance, intestinal morphology, free intestinal amino acid (AA) content, and intestinal AA transporter (AAT) mRNA levels in juvenile grass carp (Ctenopharyngodon idella). Results were displayed as follows: (1) 8% dietary glycinin decreased growth performance, inhibited intestinal growth, and caused intestinal histology damage of grass carp; (2) dietary glycinin decreased the content of free neutral AAs including Val, Ser, Tyr, Ala, Pro, and Gln in all intestinal segments, and Thr, Ile, Leu, Phe, and Gly in the MI and DI while downregulated the mRNA levels of corresponding transporters including SLC38A2, SLC6A19b, and SLC6A14 in all intestinal segments, and SLC7A5, SLC7A8, and SLC1A5 in the MI and DI. Dietary glycinin decreased the content of free basic AAs including Arg in the MI and DI and His in all intestinal segments while downregulated cationic AAT SLC7A1 mRNA levels in the MI and DI. Dietary glycinin decreased the content of free acidic AAs including Glu in all intestinal segments and Asp in the MI and DI while decreased mRNA levels of corresponding transporters including SLC1A2a in all intestinal segments and SLC1A3 in the MI and DI; (3) the digestion trial showed that basic subunits of glycinin was hard to digest in the intestine of grass carp; (4) co-administration of glutamine with glycinin partially alleviated the negative effects. Overall, glycinin decreased intestinal AA absorption capacity partly contributed by decreased AATs' mRNA levels and the indigestibility of glycinin.

A lower dose of fluorescein sodium is more suitable for confocal laser endomicroscopy: a feasibility study.

BACKGROUND AND AIMS: Image quality can be guaranteed with the conventional dosage of fluorescein sodium in probe-based confocal laser endomicroscopy (pCLE). However, yellow discoloration of the skin seriously affects daily life and simultaneously increases the risk of adverse events such as allergic reactions. The aim of this study was to test whether a lower dosage of fluorescein sodium can provide satisfactory image quality and to compare the diagnostic accuracy of gastric intestinal metaplasia (GIM) through a randomized blind controlled trial. METHODS: Consecutive patients were randomly assigned to different doses of fluorescein sodium. Image quality was determined by the endoscopists' subjective assessments and signal-to-noise ratio (SNR) assessment systems. Skin discoloration was tested using a neonatal transcutaneous jaundice detector. In addition, consecutive patients with a known or suspected diagnosis of GIM were examined by pCLE with the lower dose and the traditional dose. RESULTS: Only 0.01 mL/kg dose of 10% fluorescein sodium led to a significant decrease in image quality (P < .05), and a dose of 0.02 mL/kg had the highest SNR value (P < .05). There were no significant differences in skin discoloration between the 0.01 mL/kg and 0.02 mL/kg doses (P = .148) and no statistical difference in the diagnostic accuracy of pCLE for GIM between the 0.02 mL/kg and 0.10 mL/kg doses (P > .05). The kappa values for the correlation between pCLE and histopathology were 0.867 (95% confidence interval, 0.782-0.952) and 0.891 (95% confidence interval, 0.811-0.971). CONCLUSIONS: The 0.02 mL/kg dose of 10% fluorescein sodium seems to be the best dose for pCLE in the upper GI tract, with comparable image quality with the conventional dose and insignificant skin discoloration. This dose is also very efficient for the diagnosis of GIM.

Toxicity of Six Insecticides on Codling Moth (Lepidoptera: Tortricidae) and Effect on Expression of Detoxification Genes.

The codling moth, Cydia pomonella (L.), is a key worldwide fruit pest that has evolved high levels of resistance to almost all classes of conventional insecticides. Neonicotinoids, a new reduced-risk biorational insecticide class, have remained an effective control approach. In this study, the toxicity and sublethal effect of conventional and reduced-risk biorational insecticides on transcripts abundance of three detoxification genes in codling moth were determined. Bioassays on a codling moth laboratory strain suggested that acetamiprid had the highest oral toxicity against the third-instar larvae compared with the other five pesticides. Results also indicated that acetamiprid exhibits long-term efficacy against codling moth even at 120 h post feeding. Real-time quantitative polymerase chain reaction showed that the detoxification genes CYP9A61, CpGST1, and CpCE-1 were differentially induced or suppressed by deltamethrin, cypermethrin, methomyl, carbaryl, and imidacloprid, depending on the type of insecticides; in contrast, no significant difference in CYP9A61, CpGST1, and CpCE-1 expressions were observed after acetamiprid exposure, when compared with the control. These results suggest that the reduced-risk biorational insecticide acetamiprid is an effective insecticide with no induction of detoxification genes and can be integrated into the management of codling moth.

Isolation and functional analysis of the pathogenicity-related gene VdPR3 from Verticillium dahliae on cotton.

The fungal plant pathogen Verticillium dahliae is the causal agent of vascular wilt, a disease that can seriously diminish cotton fiber yield. The pathogenicity mechanism and the identity of the genes that interact with cotton during the infection process still remain unclear. In this study, we investigated the low-pathogenic, non-microsclerotium-producing mutant vdpr3 obtained in a previous study from the screening of a T-DNA insertional library of the highly virulent isolate Vd080; the pathogenicity-related gene (VdPR3) in wild-type strain Vd080 was cloned. Knockout mutants (ΔVdPR3) showed lower mycelium growth and obvious reduction in sporulation ability without microsclerotium formation. An evaluation of carbon utilization in mutants and wild-type isolate Vd080 demonstrated that mutants-lacking VdPR3 exhibited decreased cellulase and amylase activities, which was restored in the complementary mutants (ΔVdPR3-C) to levels similar to those of Vd080. ΔVdPR3 postponed infectious events in cotton and showed a significant reduction in pathogenicity. Reintroduction of a functional VdPR3 copy into ΔVdPR3-C restored the ability to infect cotton plants. These results suggest that VdPR3 is a multifunctional gene involved in growth development, extracellular enzyme activity, and virulence of V. dahliae on cotton.

The Toxicology and Biochemical Characterization of Cantharidin on Cydia pomonella.

Cantharidin, a natural toxin produced by beetles in the families Meloidae and Oedemeridae, reported to be toxic to some pests, is being developed as a biopesticide in China. This study evaluates the toxicity and biochemical characterization of cantharidin on the codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), an economically important fruit pest, under both laboratory and field conditions. Laboratory dose response bioassays showed that the LC50 value of cantharidin against neonate larvae was 0.057 mg ml(-1). Exposure of the larvae to 0.024 and 0.057 mg ml(-1) of cantharidin resulted in significant reduction in larval body weight. Neonate larvae exposed to LC10 of cantharidin showed increased glutathione S-transferase activity and significantly reduced the carboxylesterase and cytochrome P450-dependent mixed-function oxidase activities. Results also showed 16 and 25% ovicidal activity at concentrations of 0.057 and 0.14 mg ml(-1) of cantharidin, respectively. Field trials demonstrated cantharidin has a significant effect on both the first and second generations of C. pomonella larvae, but it exhibits a lower control efficiency than the chemical reference emamectin benzoate. Cantharidin may be considered a valuable tool for the control of codling moth.

Effectiveness of 12 Insecticides to a Laboratory Population of Cydia pomonella (Lepidoptera: Tortricidae) Newly Established in China.

The codling moth Cydia pomonella (L.) is an economically important fruit pest that has spread rapidly from its original site in Xinjiang to other northwestern regions of China. Insecticides are widely used to control this pest but its invasion has never been completely stopped. The aim of this study was to establish a laboratory population of the codling moth occurring in China, to investigate the effectiveness of 12 conventional insecticides to this laboratory population, and to recommend the discriminating doses for use in resistance monitoring. The laboratory population was generally similar to other laboratory strains although parameters such as survival rate and larval duration were low when compared with field populations. Toxicity varied among the insecticides tested with LC50 values ranging from 0.016 mg/l for emamectin benzoate to 55.77 mg/l for chlorbenzuron. Discriminating dose levels were determined from dose-mortality reference curves for the detection of resistance in field populations. Effectiveness of 12 insecticides to a laboratory population of codling moth in China was evaluated for the first time. This can be integrated into resistance management strategies, especially in orchards with a history of frequent insecticides applications, in order to monitor or decrease insecticide resistance in the future.

Review of the genus Sobrala Dworakowska (Hemiptera: Cicadellidae: Typhlocybinae: Alebrini) with description of four new species and a new record from China.

The genus Sobrala Dworakowska is reported for the first time in China and a key to known species of the genus is provided. Four new species, Sobrala acanthophylla, S. dactylina, S. lamellaris and S. quadrilatera spp. nov. from China are described and illustrated.

Key amino acid associated with acephate detoxification by Cydia pomonella carboxylesterase based on molecular dynamics with alanine scanning and site-directed mutagenesis.

Insecticide-detoxifying carboxylesterase (CE) gene CpCE-1 was cloned from Cydia pomonella. Molecular dynamics (MD) simulation and computational alanine scanning (CAS) indicate that Asn 232 in CpCE-1 constitutes an approximate binding hot-spot with a binding free energy difference (ΔΔGbind) value of 3.66 kcal/mol. The catalytic efficiency (kcat/km) of N232A declined dramatically, and the half inhibitory concentrations (IC50) value increased by more than 230-fold. Metabolism assay in vitro reveals that the acephate could be metabolized by wild CpCE-1, whereas N232A mutation is unable to metabolize the acephate, which suggests that the hot-spot Asn 232 is a crucial residue for acephate metabolism. Mutation detection suggests that low frequency of Asn 232 replacement occurred in Europe field strains. Our MD, CAS, site-directed mutagenesis, and metabolism studies introduce a new amino acid residue Asn 232 involved in the metabolism of the acephate with CpCE-1, and this method is reliable in insecticide resistance mechanism research and prediction of key amino acids in a protein which is associated with specific physiological and biochemical functions.

New species and records of Asymmetrasca (Hemiptera: Cicadellidae: Typhlocybinae: Empoascini) from China and name changes in Empoasca (Matsumurasca).

Six new species of Asymmetrasca Dlabola are described from China: A. dahaituoensis, A. helica, A. hypercurvata, A. liaoensis, A. reflexilis, A. uncus spp. nov. In addition, A. cienka Dworakowska, 1982, n. comb.; A. decedens Paoli, 1932 and A. sakaii Dworakowska 1971, n. comb., are recorded from China for the first time. The following additional new combinations from Empoasca are also proposed: A. cisiana (Dworakowska, 1971); A. kaicola (Dworakowska, 1982), A. lutowa (Dworakowska, 1971), A. mona (Dworakowska, 1994), A. nipponica (Dworakowska 1982), A. rybiogon (Dworakowska, 1971), and A. uniprossicae (Sohi, 1977). Empoasca (Empoasca) kishtwarensis Sharma, 1984 is proposed as a new synonym of Asymmetrasca kaicola (Dworakowska, 1982). Habitus photos and illustrations of the male genitalia of the new species and a key to Chinese species are provided. New names are proposed to replace two junior homonyms in Empoasca (Matsumurasca).

Survey and clinical considerations of gender identity in lower primary school children.

BACKGROUND: Gender consciousness directly affects the development of gender identity, which is a continuous and lifelong process. Meanwhile, hospitalization is a part of many children's lives and has an impact on their gender development. AIM: To investigate the current situation of gender identity in lower primary school children by conducting a survey of 202 hospitalized children in the lower grades and to provide a theoretical basis and foundation for the cultivation of gender identity and medical treatment of children based on the results. This study aims to inspire clinical medical staff to scientifically and reasonably arrange hospital wards for lower primary school children and pay attention to gender protection during the medical treatment process and to help children shape a unified and clear gender identity, which will enable them to better integrate into society and promote their personality development. METHODS: The gender consciousness scale for elementary and middle school students was used for the survey. RESULTS: Gender identity was already present in lower primary school children. The children's gender roles and gender equality consciousness were strong, exceeding the critical value, but their gender characteristics, gender identity, and gender ideal consciousness were weak. Children aged 6 had the weakest gender identity, and girls had significantly stronger gender identity than boys. CONCLUSION: Gender identity is already present in lower primary school children, providing a basis and inspiration for the cultivation of gender identity and medical treatment of lower primary school children. Clinical medical staff should be aware of and understand these results and should scientifically and reasonably arrange hospital wards for lower primary school children.

Cantharidin Impedes Activity of Glutathione S-Transferase in the Midgut of Helicoverpa armigera Hübner.

Previous investigations have implicated glutathione S-transferases (GSTs) as one of the major reasons for insecticide resistance. Therefore, effectiveness of new candidate compounds depends on their ability to inhibit GSTs to prevent metabolic detoxification by insects. Cantharidin, a terpenoid compound of insect origin, has been developed as a bio-pesticide in China, and proves highly toxic to a wide range of insects, especially lepidopteran. In the present study, we test cantharidin as a model compound for its toxicity, effects on the mRNA transcription of a model Helicoverpa armigera glutathione S-transferase gene (HaGST) and also for its putative inhibitory effect on the catalytic activity of GSTs, both in vivo and in vitro in Helicoverpa armigera, employing molecular and biochemical methods. Bioassay results showed that cantharidin was highly toxic to H. armigera. Real-time qPCR showed down-regulation of the HaGST at the mRNA transcript ranging from 2.5 to 12.5 folds while biochemical assays showed in vivo inhibition of GSTs in midgut and in vitro inhibition of rHaGST. Binding of cantharidin to HaGST was rationalized by homology and molecular docking simulations using a model GST (1PN9) as a template structure. Molecular docking simulations also confirmed accurate docking of the cantharidin molecule to the active site of HaGST impeding its catalytic activity.

Cloning and expression analysis of a muscarinic cholinergic receptor from the brain of ant, Polyrhachis vicina.

Muscarinic acetylcholine receptors (mAchRs) are the predominant cholinergic receptors in the central and peripheral nervous systems of animals. They also have been found in various insect nervous systems. In this article, a full-length cDNA of a pupative mAchR (PmAchR) was obtained from the brains of ant Polyrhachis vicina by homology cloning in combination with rapid amplification of cDNA ends. PmAchR encodes a 599-amino acid protein that exhibits a high degree of homology with other mAchRs. Real-time quantitative RT-PCR analysis showed that PmAchR is differentially expressed in the brains of workers, males, and females. By in situ hybridization, it is revealed that PmAchR is widely expressed in different soma clusters of the brain, including the mushroom bodies, the antennal lobes, as well as the optic lobes (OL), and the most intensely staining is found in Kenyon cells. Nonetheless, there are more positive nerve fibers in the OL of males' brains than in females' and workers' brains.

[Association between aggressive behaviors and COMT Val158Met and 5-HTTLPR polymorphisms in children].

OBJECTIVE: To investigate the association between aggressive behaviors and catechol-O-methyltransferase (COMT) single nucleotide polymorphism at position 158 from a valine to a methionine (Val158Met) as well as serotonin (5-HT) transporter gene linked polymorphic region (5-HTTLPR) in children. METHODS: A total of 68 children who were exposed to domestic violence were recruited. The frequencies of genotypes and alleles of COMT Val158Met and 5-HTTLPR were examined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques. A comparison was conducted between 24 children with high scores of aggressive problems and 44 control children with low scores of aggressive problems according to Child Behavior Checklist (for parents).ResultsThere were no significant differences in genotypes of COMT Val158Met (χ2=1.612, P=0.447) and 5-HTTLPR (χ2=1.807, P=0.405) between the two groups. There were also no significant differences in the frequencies of alleles of COMT Val158Met (χ2=1.648, P=0.119) and 5-HTTLPR(χ2=0.403, P=0.527) in the two groups. CONCLUSIONS: COMT Val158Met and 5-HTTLPR might not be the susceptible genes of children's aggression, suggesting that children's aggressive behaviors might be affected by multivariate factors.

Complete mitochondrial genome of Lelecella limenitoides (Lepidoptera: Nymphalidae: Apaturinae) and phylogenetic implication.

The length of Lelecella limenitoides complete mitogenome was 15,203 bp and contained the typical gene arrangement, base composition, and codon usage found in other related species. The overall base composition exhibited obvious anti-G (7.5%) and AT bias (81.6%). The initiation codons of all PCGs were typical ATN (ATA/ATG/ATT), and the termination codons were TAA, TAG, or incomplete stop codon T-. All tRNAs could be folded into typical cloverleaf secondary structures, except tRNASer (AGN). Phylogenetic analysis showed that L. limenitoides was clustered with the clade of Sasakia, Euripus, and Apatura.

Convergent Synthesis of Dihydropyrans from Catalytic Three-Component Reactions of Vinylcyclopropanes, Diazoesters, and Diphenyl Sulfoxide.

A novel Rh(I)/La(III) cocatalytic three-component reaction of vinylcyclopropanes, diazoesters, and diphenyl sulfoxide has been developed. The reaction gives polysubstituted dihydropyrans as the reaction products. Mechanistic studies indicate that isomerization of vinylcyclopropanes gives conjugated dienes, which then undergo [4 + 2]-cycloaddition with vicinal tricarbonyl compounds generated by oxygen atom transfer from diphenyl sulfoxide to diazoesters.

Rhodium-Catalyzed ortho-Selective Carbene C-H Insertion of Unprotected Phenols Directed by a Transient Oxonium Ylide Intermediate.

ortho-Selective carbene C-H insertion of unprotected phenols is achieved with dimethyl diazomalonate under the catalysis of [Rh(COD)Cl]2. After the C-H insertion, subsequent cyclization and electrophilic addition of another carbene molecule affords tris-carboxylate-substituted 2-benzofuranones as the final reaction products. The enantioselective variant has been developed with the use of chiral diene ligands. Mechanistic experiments indicate that a transient oxonium ylide directing group might be responsible for the regiocontrol at the C-H activation step.

Clinical differences between late-onset and early-onset chronically hospitalized elderly schizophrenic patients in Taiwan.

OBJECTIVES: To examine the clinical differences between late-onset schizophrenia (LOS) and early-onset schizophrenia (EOS) in Taiwanese elderly chronic hospitalized schizophrenic patients. METHODS: By using a cross-sectional study method, we investigated all the hospitalized elderly schizophrenic patients in a general hospital's psychiatric ward during July-September 2007. All the subjects matched DSM-IV-TR diagnostic criteria and were aged 60 years or above. A total of 52 subjects were enrolled (LOS = 23, EOS = 29). Demographic data, illness history, and antipsychotic treatment record were documented; the positive and negative syndrome scale (PANSS), Center for Epidemiological Studies Depression scale (CES-D), mini-mental status examination (MMSE), activities of daily living rating scale for psychiatric patients (ADLRS), community self-sufficiency test (CST), clinical global impression (CGI), and general assessment of functioning scale (GAF) were administered. RESULTS: In our samples, a lower educational level was found to be more common in late-onset patients. In LOS, there was no significant increase in the severity of PANSS psychopathology except for greater thought disorder symptoms. Both LOS and EOS patients had similar cognitive and functioning impairment with poor global outcomes. There was a trend of low antipsychotic drug use in LOS. CONCLUSIONS: In Taiwan, elderly chronic inpatients LOS had greater thought disorders compared to findings in Western studies. Further large-scale longitudinal studies are needed to understand the factors related to these findings.