A Phytophthora infestans RXLR effector AVR8 suppresses plant immunity by targeting a desumoylating isopeptidase DeSI2.

The potato's most devastating disease is late blight, which is caused by Phytophthora infestans. Whereas various resistance (R) genes are known, most are typically defeated by this fast-evolving oomycete pathogen. However, the broad-spectrum and durable R8 is a vital gene resource for potato resistance breeding. To support an educated deployment of R8, we embarked on a study on the corresponding avirulence gene Avr8. We overexpressed Avr8 by transient and stable transformation, and found that Avr8 promotes colonization of P. infestans in Nicotiana benthamiana and potato, respectively. A yeast-two-hybrid (Y2H) screen showed that AVR8 interacts with a desumoylating isopeptidase (StDeSI2) of potato. We overexpressed DeSI2 and found that DeSI2 positively regulates resistance to P. infestans, while silencing StDeSI2 downregulated the expression of a set of defense-related genes. By using a specific proteasome inhibitor, we found that AVR8 destabilized StDeSI2 through the 26S proteasome and attenuated early PTI responses. Altogether, these results indicate that AVR8 manipulates desumoylation, which is a new strategy that adds to the plethora of mechanisms that Phytophthora exploits to modulate host immunity, and StDeSI2 provides a new target for durable resistance breeding against P. infestans in potato.

Metal Variance in Multivariate Metal-Organic Frameworks for Boosting Catalytic Conversion of CO2.

Developing efficient, low-cost, MOF catalysts for CO2 conversion at low CO2 concentrations under mild conditions is particularly interesting but remains highly challenging. Herein, we prepared an isostructural series of two-dimensional (2D) multivariate metal-organic frameworks (MTV-MOFs) containing copper- and/or silver-based cyclic trinuclear complexes (Cu-CTC and Ag-CTC). These MTV-MOFs can be used as efficient and reusable heterogeneous catalysts for the cyclization of propargylamine with CO2. The catalytic performance of these MTV-MOFs can be engineered by fine-tuning the Ag/Cu ratio in the framework. Interestingly, the induction of 10% Ag remarkably improved the catalytic efficiency with a turnover frequency (TOF) of 243 h-1, which is 20-fold higher than that of 100% Cu-based MOF (i.e., TOF = 10.8 h-1). More impressively, such a bimetallic MOF still exhibited high catalytic activity even for simulated flue gas with 10% CO2 concentration. Furthermore, the reaction mechanism has been examined through the employment of NMR monitoring experiments and DFT calculations.

Normobaric hypoxia shows enhanced FOXO1 signaling in obese mouse gastrocnemius muscle linked to metabolism and muscle structure and neuromuscular innervation.

Skeletal muscle relies on mitochondria for sustainable ATP production, which may be impacted by reduced oxygen availability (hypoxia). Compared with long-term hypoxia, the mechanistic in vivo response to acute hypoxia remains elusive. Therefore, we aimed to provide an integrated description of the Musculus gastrocnemius response to acute hypoxia. Fasted male C57BL/6JOlaHsd mice, fed a 40en% fat diet for six weeks, were exposed to 12% O2 normobaric hypoxia or normoxia (20.9% O2) for six hours (n = 12 per group). Whole-body energy metabolism and the transcriptome response of the M. gastrocnemius were analyzed and confirmed by acylcarnitine determination and Q-PCR. At the whole-body level, six hours of hypoxia reduced energy expenditure, increased blood glucose and tended to decreased the respiratory exchange ratio (RER). Whole-genome transcriptome analysis revealed upregulation of forkhead box-O (FOXO) signalling, including an increased expression of tribbles pseudokinase 3 (Trib3). Trib3 positively correlated with blood glucose levels. Upregulated carnitine palmitoyltransferase 1A negatively correlated with the RER, but the significantly increased in tissue C14-1, C16-0 and C18-1 acylcarnitines supported that ß-oxidation was not regulated. The hypoxia-induced FOXO activation could also be connected to altered gene expression related to fiber-type switching, extracellular matrix remodeling, muscle differentiation and neuromuscular junction denervation. Our results suggest that a six-hour exposure of obese mice to 12% O2 normobaric hypoxia impacts M. gastrocnemius via FOXO1, initiating alterations that may contribute to muscle remodeling of which denervation is novel and warrants further investigation. The findings support an early role of hypoxia in tissue alterations in hypoxia-associated conditions such as aging and obesity.

The Analysis of the Glycosyltransferase Gene Function From a Novel Granaticin Producer, Streptomyces Vilmorinianum. YP1.

Glycosylation is common among the synthesis of natural product and imparts the bioactivity for natural product. As for granaticin, a natural product with great bioactivity, glycosylation is an unusual sugar attachment and remains enigmatic. Orf14 in the gra cluster is the predicted glycosyltransferase but without being identified. Recently, we isolated and identified a novel granaticin producer Streptomyces vilmorinianum YP1. Orf14 gene in gra cluster of YP1 is knocked out and complemented. The instrumental analysis of the blue product synthesized by orf14-deficient mutant exhibits the none-granaticin detection and deglycosylated intermediates accumulation. The bioactivity and stability test suggests the weaker or none antibacterial activity and cytotoxicity of this blue product with greater ultraviolet stability and thermostability than granaticin and derivatives produced by YP1. All the result indicates that orf14 encodes glycosyltransferase and glycosylation played an important role in the bioactivity of granaticin. Meanwhile, the blue pigment, deglycosylated intermediates, has favorable processing characteristics. Our finding supplies the function of orf14 and glycosylation, but also indicates a promising candidate of edible blue pigment applicated in food industry.

An Au Nanofilm-Graphene/D-Type Fiber Surface Plasmon Resonance Sensor for Highly Sensitive Specificity Bioanalysis.

A highly sensitive Au-graphene structure D-type fiber surface plasmon resonance biosensor is presented in this study to specifically detect biomolecules. The method of growing graphene is employed directly on the copper, and then a gold film of optimum thickness is sputtered, and the copper foil is etched to obtain the structure. This method makes the contact closer between the gold layer and the graphene layer to improve surface plasmon resonance performance. The performance of this type of surface plasmon resonance (SPR) sensor has been previously verified both theoretically and experimentally. With the proposed Au-graphene structure D-type fiber biosensor, the SPR behaviors are obtained and discussed. In the detection of ethanol solution, a red shift of 40 nm is found between the refractive index of 1.3330 and 1.3657. By calculation, the sensitivity of the sensor we designed is 1223 nm/RIU. Besides, the proposed sensor can detect the nucleotide bonding between the double-stranded DNA helix structures. Thus, our sensors can distinguish between mismatched DNA sequences.

The Inhibition of miR-144-3p on Cell Proliferation and Metastasis by Targeting TOP2A in HCMV-Positive Glioblastoma Cells.

Glioblastoma (GBM), the most common type of primary tumor in the central nervous system, is a very aggressive brain tumor with poor prognosis and a high recurrence rate. Increasing evidence suggests that human cytomegalovirus (HCMV) infection is related to GBM and leads to GBM cell growth and metastasis. MicroRNAs are important regulators in the growth and metastasis of glioblastoma. This study aimed to demonstrate the role of miR-144-3p in HCMV-positive glioblastoma. We found that, after HCMV infection, the expression of miR-144-3p decreased, whereas the expression of TOP2A increased. Bioinformatics analyses indicated that miR-144-3p directly targets the TOP2A 3'-UTR (Untranslated Region). We discovered that the overexpression of miR-144-3p downregulated the overexpression of TOP2A and inhibited the proliferation, clone formation, and invasion of HCMV-positive glioma in vitro. Taken together, these results show that miR-144-3p inhibited growth and promoted apoptosis in glioma cells by targeting TOP2A.

Long non-coding RNA PVT1 promotes glycolysis and tumor progression by regulating miR-497/HK2 axis in osteosarcoma.

Cancer cells usually utilize glucose as a carbon source for aerobic glycolysis, a phenomenon known as the Warburg effect. Increasing studies have shown that PVT1 (a long non-coding RNA, lncRNA) functions as critical regulator of multiple cancers. However, it remains totally unknown whether and how PVT1 regulates glucose metabolism in OS cells. In this study, we found that the expression of PVT1 were specifically increased in OS cells and tissues, and the upregulated PVT1 indicated poor prognosis. Glucose uptake, lactate production, and the expression of HK2 in OS cells were increased by overexpression of PVT1, while decreased by PVT1 knockdown. We further fund that PVT1 acted as molecular sponge to repress miR-497. Inhibition of miR-497 promoted glucose consumption and lactate production, phenomenon could be reversed by PVT1 silencing. Moreover, HK2 was a direct target of miR-497 and overexpression of HK2 attenuated the suppressive effect of miR-497 on glycolysis. Functionally, knockdown of PVT1 exerted tumor-suppressive effect by suppressing cell proliferation, cell cycle progression, and invasion in vitro, whereas miR-497 inhibitor partially abolished the inhibition effect of si-PVT1. Overexpression of HK2 attenuated the miR-497 induced inhibition of cell growth and motility. Taken together, these findings suggested that PVT1 contributes to OS cell glucose metabolism, cell proliferation, and motility through the miR-497/HK2 pathway, and revealed a novel relation between lncRNA and the alteration of glycolysis in OS cells.

Investigation of Growth Phase-Dependent Acid Tolerance in Bifidobacteria longum BBMN68.

The underlying mechanisms imparting the growth phase-dependent acid tolerance have not been extensively investigated. In this study, we compared the acid resistance of the Bifidobacterium longum strain BBMN68 from different growth phases at lethal pH values (pH 2.5, 3.0, and 3.5), and analyzed the activity of H(+)-ATPase, the composition of fatty acids, and the mRNA abundance of ffh, uvrA, recA, lexA, groES, and dnaK in cells from different growth phases. The results indicated that the survival rates of cells from early stationary (ES) and late stationary (LS) growth phases at lethal pH values were significantly higher than those of exponential growth phase cells. Our findings indicated that by inducing a continuously auto-acidizing environment during cell growth, the acid resistance of ES and LS cells was strengthened. The higher activity of H(+)-ATPase, the decrease in unsaturated fatty acids, and the increased expression of genes involved in DNA repair and protein protection in the cells in stationary growth phase were all implicated in the significantly increased acid resistance of ES and LS cells compared with exponential growth phase cells of the B. longum strain BBMN68.

Transforming growth factor beta signaling in hepatocytes participates in steatohepatitis through regulation of cell death and lipid metabolism in mice.

UNLABELLED: Transforming growth factor beta (TGF-ß) signaling activates Smad- and TGF-ß-activated kinase 1 (TAK1)-dependent signaling to regulate cell survival, proliferation, fibrosis, and tumorigenesis. The effects of TGF-ß signaling on metabolic syndrome, including nonalcoholic fatty liver disease, remain elusive. Wild-type (WT) and hepatocyte-specific TGF-ß receptor type II-deficient (Tgfbr2ΔHEP) mice were fed a choline-deficient amino acid (CDAA)-defined diet for 22 weeks to induce NASH. WT mice fed a CDAA diet displayed increased activation of Smad2/3 and had marked lipid accumulation, inflammatory cell infiltration, hepatocyte death, and fibrosis; in comparison, Tgfbr2ΔHEP mice fed a CDAA diet had suppressed liver steatosis, inflammation, and fibrosis. Both palmitate-induced steatotic hepatocytes and hepatocytes isolated from WT mice fed a CDAA diet had increased susceptibility to TGF-ß-mediated death. TGF-ß-mediated death in steatotic hepatocytes was inhibited by silencing Smad2 or blocking reactive oxygen species (ROS) production and was enhanced by inhibiting TAK1 or nuclear factor kappa B. Increased hepatic steatosis in WT mice fed a CDAA diet was associated with the increased expression of lipogenesis genes (Dgat1 and Srebp1c), whereas the decreased steatosis in Tgfbr2ΔHEP mice was accompanied by the increased expression of genes involved in ß-oxidation (Cpt1 and Acox1). In combination with palmitate treatment, TGF-ß signaling promoted lipid accumulation with induction of lipogenesis-related genes and suppression of ß-oxidation-related genes in hepatocytes. Silencing Smad2 decreased TGF-ß-mediated lipid accumulation and corrected altered gene expression related to lipid metabolism in hepatocytes. Finally, we confirmed that livers from patients with nonalcoholic steatohepatitis (NASH) displayed phosphorylation and nuclear translocation of Smad2/3. CONCLUSIONS: TGF-ß signaling in hepatocytes contributes to hepatocyte death and lipid accumulation through Smad signaling and ROS production that promote the development of NASH.

Transforming growth factor-ß signaling in hepatocytes promotes hepatic fibrosis and carcinogenesis in mice with hepatocyte-specific deletion of TAK1.

BACKGROUND & AIMS: Transforming growth factor (TGF)-ß-activated kinase 1 (TAK1) is activated in different cytokine signaling pathways. Deletion of Tak1 from hepatocytes results in spontaneous development of hepatocellular carcinoma (HCC), liver inflammation, and fibrosis. TGF-ß activates TAK1 and Smad signaling, which regulate cell death, proliferation, and carcinogenesis. However, it is not clear whether TGF-ß signaling in hepatocytes, via TGF-ß receptor-2 (Tgfbr2), promotes HCC and liver fibrosis. METHODS: We generated mice with hepatocyte-specific deletion of Tak1 (Tak1ΔHep), as well as Tak1/Tgfbr2DHep and Tak1/Smad4ΔHep mice. Tak1flox/flox, Tgfbr2ΔHep, and Smad4ΔHep mice were used as controls, respectively. We assessed development of liver injury, inflammation, fibrosis, and HCC. Primary hepatocytes isolated from these mice were used to assess TGF-ß-mediated signaling. RESULTS: Levels of TGF-ß, TGF-ßR2, and phospho-Smad2/3 were increased in HCCs from Tak1ΔHep mice, which developed liver fibrosis and inflammation by 1 month and HCC by 9 months. However, Tak1/Tgfbr2ΔHep mice did not have this phenotype, and their hepatocytes did not undergo spontaneous cell death or compensatory proliferation. Hepatocytes from Tak1ΔHep mice incubated with TGF-ß did not activate p38, c-Jun N-terminal kinase, or nuclear factor-κB; conversely, TGF-ß-mediated cell death and phosphorylation of Smad2/3 were increased, compared with control hepatocytes. Blocking the Smad pathway inhibited TGF-ß-mediated death of Tak1-/- hepatocytes. Accordingly, disruption of Smad4 reduced the spontaneous liver injury, inflammation, fibrosis, and HCC that develops in Tak1ΔHep mice. Levels of the anti-apoptotic protein Bcl-xL, ß-catenin, connective tissue growth factor, and vascular endothelial growth factor were increased in HCC from Tak1ΔHep mice, but not in HCCs from Tak1/Tgfbr2ΔHep mice. Injection of N-nitrosodiethylamine induced HCC formation in wild-type mice, but less in Tgfbr2ΔHep mice. CONCLUSIONS: TGF-ß promotes development of HCC in Tak1ΔHep mice by inducing hepatocyte apoptosis and compensatory proliferation during early phases of tumorigenesis, and inducing expression of anti-apoptotic, pro-oncogenic, and angiogenic factors during tumor progression.

Highly sensitive humidity sensor based on composite film of partially reduced graphene oxide and bacterial cellulose.

Breathing is an important physiological activity of human body, which not only reflects the state of human movement, but also is one of the important health indicators. Breathing can change the concentration of water molecules, so monitoring humidity has gradually become a hot topic in modern research. In this study, a humidity sensing composite film with high sensitivity and short response time was made by using the mixture of graphene oxide (GO) and bacterial cellulose (BC) with simple dry film-forming method. L-ascorbic acid was used as reducing agent to reduce GO and improve the conductivity of GO/BC composite film (BG). The influence of different BC contents and the different reduction degree on the resistance change rate of composite film was investigated in details. The maximum resistance change rate of partially reduced BG humidity sensitive composite film reached up to 94%, and the response and recovery time were 13 s and 47 s respectively. Furthermore, the sensor shows obvious resistance change in noncontact sensing test and different breathing states. This kind of humidity sensitive film with fast response and high sensitivity has great potential in human health monitoring and noncontact sensing, and is of great significance in promoting health detection and intelligent life.

Inflammatory Pathogenesis of Post-stroke Depression.

Post-stroke depression (PSD) is a complex mood disorder that emerges in individuals following a stroke, characterized by the development of depressive symptoms. The pathogensis of PSD is diverse, with inflammation playing a vital role in its onset and progression. Emerging evidence suggests that microglial activation, astrocyte responses, nuclear factor κB(NF-κB) signaling, dysregulation of the hypothalamic pituitary adrenal (HPA) axis, alterations in brain-derived neurotrophic factor (BDNF) expression, neurotransmitter imbalances, adenosine triphosphate (ATP) and its receptors and oxidative stress are intricately linked to the pathogenesis of PSD. The involvement of inflammatory cytokines in these processes highlights the significance of the inflammatory pathway. Integrating these hypotheses, the inflammatory mechanism offers a novel perspective to expand therapeutic strategies for PSD.

Targeted delivery of Fc-fused PD-L1 for effective management of acute and chronic colitis.

The PD-1/PD-L1 pathway in mucosal immunity is currently actively explored and considered as a target for inflammatory bowel disease (IBD) treatment. However, systemic PD-L1 administration may cause unpredictable adverse effects due to immunosuppression. Here we show that reactive oxygen species (ROS)-responsive nanoparticles enhance the efficacy and safety of PD-L1 in a mouse colitis model. The nanoparticles control the accumulation and release of PD-L1 fused to Fc (PD-L1-Fc) at inflammatory sites in the colon. The nanotherapeutics shows superiority in alleviating inflammatory symptoms over systemic PD-L1-Fc administration and mitigates the adverse effects of PD-L1-Fc administration. The nanoparticles-formulated PD-L1-Fc affects production of proinflammatory and anti-inflammatory cytokines, attenuates the infiltration of macrophages, neutrophils, and dendritic cells, increases the frequencies of Treg, Th1 and Tfh cells, reshapes the gut microbiota composition; and increases short-chain fatty acid production. In summary, PD-L1-Fc-decorated nanoparticles may provide an effective and safe strategy for the targeted treatment of IBD.

Lipidomics analysis facilitate insight into the molecular mechanisms of urate nephropathy in a gout model induced by combination of MSU crystals injection and high-fat diet feeding.

Renal injury is one of the most common clinical manifestations of patients with hyperuricaemia/gout. The precise pathophysiological mechanism(s) for the renal injury is still unknown. Furthermore, it is also unclear whether the clinical therapies (e.g., colchicine and febuxostat) could prevent its progression or not. Lipids are involved in almost all of important biological processes and play critical roles in maintaining the renal functions. Herein, shotgun lipidomics was performed for class-targeted lipid analysis of cellular lipidomes in renal tissue of a gouty model induced by combination of monosodium urate crystals injection and high-fat diet feeding with/without treatment with either colchicine or febuxostat. Serum uric acid (UA), proinflammatory cytokines (i.e., TNF-α and IL-6), xanthine oxidase activity, footpad swelling, and pain threshold were determined to evaluate the gouty severity. Renal histopathological changes, blood urea nitrogen, creatinine, and kidney index were used to reflect renal injury. Lipidomics analysis revealed that altered triacylglycerol (TAG) profile, impaired mitochondrial function resulted by decreased tetra 18:2 cardiolipin, reduced 4-hydroxyalkenal (HNE) species, and elevated lysophospholipids were already present in the kidneys at early stage of renal injury, probably contributing to its occurrence and development. In addition to significantly reduce the UA level and relief the gouty severity, treatment with either colchicine or febuxostat could restore HNE bioavailability, thereby delaying the progression of renal injury. However, both of them could not recover the altered TAG profile and the impaired mitochondrial function, indicating that treatment with either of them could not completely prevent the development of renal injury in the gouty model.

Lipidomics characterized TAG biosynthesis of developing kernels in three walnut cultivars in Xinjiang region.

Walnut oil with very high proportion of polyunsaturated fatty acids exhibits many health beneficial effects. We hypothesized that the oil composition is led by a special pattern/mechanism for triacylglycerol (TAG) biosynthesis as well as accumulation in walnut kernel during embryo development. To test this hypothesis, shotgun lipidomics was performed for class-targeted lipid analysis (including TAG, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylglycerol, phosphatidylinositol, and lysophosphatidylcholine species) in walnut kernels from three cultivar collected at three critical stages of embryo development. The results indicated that TAG synthesis in the kernel happened before 84 days after flowering (DAF) and was significantly enhanced between 84 and 98 DAF. Moreover, TAG profile was changing along with DAFs due to the increased composition of 18:1 FA in TAG pool. Moreover, lipidomics also demonstrated that the enhanced acyl editing was responsible for the flux of FA through phosphatidylcholine for eventual TAG synthesis. Therefore, TAG biosynthesis in walnut kernel was characterized directly from lipid metabolism.

Effects of food emulsifiers on high fat-diet-induced obesity, intestinal inflammation, changes in bile acid profile, and liver dysfunction.

Obesity has become one of the most prevalent health concerns of our time. A long-term high-fat diet is closely related to obesity. Food emulsifiers are incorporated into high-fat foods to enhance the texture and stability. Whether food emulsifiers exacerbate obesity and metabolic disorders induced by a high-fat diet remains unclear. This study aimed to investigate the effects of polysorbate-80 (P80) and polyglycerol polyricinoleate (PGPR) on lipid metabolism, bile acid profile, and gut microbiota in normal and high-fat-diet-induced obesity in mice. The results of this study showed that P80 and PGPR had little effect on body weight but significantly increased epididymal-fat weight, total energy intake, and blood lipid levels. P80 and PGPR stimulated colon inflammation and improved the expression of inflammatory factors in the colon and liver significantly. P80 and PGPR changed the bile acid profile. However, P80 and PGPR did not aggravate inflammation, obesity and alter bile acid profile by altering the composition of the gut microbiota. The results of this study provide an experimental reference for the rational use of food additives and the adjustment of dietary structure, which are important and have application value.

Tuning the Catalytic Activity of Covalent Metal-Organic Frameworks for CO2 Cycloaddition Reactions.

The development of efficient, recyclable and low-cost heterogeneous catalysts for conversion of carbon dioxide (CO2 ) into epoxides is highly desired, yet remain a challenge. Herein, we have prepared three two-dimensional (2D) copper(I) cyclic trinuclear units (Cu(I)-CTUs) based covalent metal-organic frameworks (CMOFs), namely JNM-13, JNM-14, and JNM-15, via a one-pot reaction by combination of coordination and dynamic covalent chemistry. Among them, JNM-15 contained the highest density of copper catalytic sites, and exhibited the highest capacity for adsorption of CO2 . More interestingly, JNM-15 delivered the highest catalytic activity for cycloaddition of CO2 to epoxides with good yields (up to 99 %), good substrate compatibility (11 examples) and reusability (four catalytic cycles) under mild condition.

Exposure to multiple pyrethroid insecticides affects ovarian follicular development via modifying microRNA expression.

Pyrethroids, a class of insecticides widely used in agriculture and residential pest control, have been considered as endocrine-disrupting chemicals (EDCs). Our previous epidemiological study reported a positive association of urinary levels of pyrethroid metabolites with the risk of primary ovarian insufficiency in women, suggesting that pyrethroid exposure may be a potential risk factor for female ovarian health. In this study, female mice at gestational, lactational or peripubertal stages were exposed to eight most commonly used pyrethroids at the doses of acceptable daily intake (ADI) recommended by the World Health Organization (WHO). Gestational exposure to eight pyrethroids at ADI doses led to a significant decrease in the number of primary follicles in female offspring on postnatal day (PND) 3, and an increase in the number of atretic follicles and granulosa cell apoptosis, as well as lower estrogen and higher follicle-stimulating hormone (FSH) levels in adult female offspring. Lactational and peripubertal exposure to pyrethroid mixture had no significant effects on follicular development and ovarian functions. The data of high-throughput microRNA (miRNA) sequencing showed that 23 miRNAs were differentially expressed in the ovaries of female offspring mice on PND 1 after gestational exposure to pyrethroid mixture. The results of qPCR confirmed that miR-152-3p, miR-450b-3p and miR-196a-5p were significantly upregulated in the neonatal ovaries in the exposed group. The bioinformatic analysis indicates that the modification of the expression of ovarian miRNAs by pyrethroid exposure may disrupt the key biological processes (such as mRNA processing) and major signaling pathways (such as PI3K/Akt pathway, adipocytokine pathway and GnRH pathway) governing follicular development and ovarian functions. This study first reported that gestational exposure of female mice to multiple pyrethroids at the recommended human safe doses had irreversible adverse effects on the ovaries in female offspring in adulthood through regulating the expression of miRNAs during early developmental stages.

Lifelong exposure to pyrethroid insecticide cypermethrin at environmentally relevant doses causes primary ovarian insufficiency in female mice.

Pyrethroids are a class of widely used insecticides. Our recent epidemiological study of Chinese women reported that pyrethroid exposure was positively associated with the risk of primary ovarian insufficiency (POI). In this study, we utilized cypermethrin (CP), the most frequently detected pyrethroid in the environment, to recognize how lifelong and low-dose exposure to pyrethroids affects ovarian functions and the underlying mechanism(s). Female mice were exposed to CP at doses of human dietary intake of 6.7 µg/kg/day, an acceptable daily intake (ADI) of 20 µg/kg/day, or the chronic reference dose (RfD) of 60 µg/kg/day, starting from gestational day 0.5 until 44-week-old. We assessed effects on fertility, serum hormone levels, ovarian follicular development and ovarian transcriptomic profiles. Chronic exposure to CP at doses of ADI and RfD caused a significant reduction in the size of the primordial follicle pool on postnatal day (PND) 5 and the number of all types of follicles in 44-week-old mice, lower estrogen and higher gonadotropin levels, as well as decreased fertility. Significant increase in apoptosis and decrease in cell proliferation were observed in CP-exposed ovarian follicles from PND 5 and 44-week-old mice. Ovarian transcriptomic data showed that the pro-apoptotic protein BMF and the cell cycle inhibitor p27 were significantly up-regulated in CP-exposed ovaries. Cyp17a1, Cyp19a1 and Hsd17b1 genes involved in the key steps of steroidogenesis were down-regulated in the ovaries of female mice exposed to CP. This study first reported that lifelong exposure to CP at doses of ADI or RfD caused an ovarian phenotype similar to human POI in female mice and provided a mechanistic explanation. Our findings suggest that lifelong exposure to pyrethroids of low doses, which are recommended as 'safe' dosages, may have a significant impact on the ovarian health of female mammals and humans.

Expressional and functional comparisons of five clustered odorant binding proteins in the brown marmorated stink bug Halyomorpha halys.

Odorant-binding proteins (OBPs) play essential roles in the functioning of insect peripheral olfactory systems. To fully understand the olfactory roles of OBPs in Halyomorpha halys, an important invasive pest found worldwide, we studied the expression and functional characterization of five OBP-associated genes from H. halys that are clustered in the genome. The tissue distribution of the OBP gene cluster suggests that these genes were enriched in nymph and adult antennae, indicating their possible involvement in the chemosensory process. The different expression levels of the five OBPs in nymph and adult antennae suggest that this gene cluster is regulated independently. Ligand-binding experiments have shown similar specificities of these five OBPs towards several organic compounds, including the alarm pheromone of H. halys (trans-2-decenal), the aggregation pheromone of Plautia stali (methyl (2E, 4E, 6Z)-decatrienoate), and plant volatile compounds (e.g., cis-3-hexenyl benzoate and ß-ionone). In particular, trans-2-dodecenal, an alarm pheromone analog, exhibited high affinity to the five OBP proteins and alarm pheromone activity towards H. halys. Thus, this OBP cluster may mediate the response of stink bugs to the both the alarm pheromone and host-related volatiles and could be an interesting target to design novel olfactory regulators for the management of H. halys infestations.