Gestational exposure to perfluoroalkyl substances is associated with placental DNA methylation and birth size.

DNA methylation is one potential mechanism for the effects of gestational exposure to perfluoroalkyl substances (PFASs) on fetal growth. We investigated 180 pregnant women who participated in a cohort study conducted in Tangshan City, Northern China, and determined the concentrations of 11 PFASs and the methylation of two genes related to fetal growth [insulin-like growth factor 2 (IGF2) and nuclear receptor subfamily 3 group C member 1 (NR3C1)] and one surrogate marker for global methylation [long interspersed nuclear element-1 (LINE-1)] in placenta tissue. Multiple linear regression analysis was performed to examine the associations of log transformed PFASs with the DNA methylation and birth size. Weighted quantile sum regression was used to determine the mixture effect of PFASs. After adjusting for potential confounders, perfluorooctane sulfonate (PFOS) was negatively associated with the overall methylation of LINE-1. PFASs mixture was negatively associated with the methylation of all CpG loci of LINE-1 and overall methylation of NR3C1. Perfluorootanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), and the PFASs mixture showed negative associations with head circumference. After stratified by newborns' sex, PFOA, PFNA and the PFASs mixture was negatively associated with overall methylation of LINE-1 only in the male subgroup and the methylation of all CpG loci of LINE-1 was negatively associated with ponderal index only in the female subgroup. The interaction of newborns' sex with PFOS and PFOA on overall methylation of IGF2 was statistically significant and so was the interaction of sex with PFOS on overall methylation of LINE-1. These findings suggested that intrauterine exposure to PFASs affected placental DNA methylation and reduced fetal growth, which might be modified by sex.

Exposure to perfluoroalkyl substances was associated with estrogen homeostasis in pregnant women.

Previous studies have suggested that perfluoroalkyl substances (PFASs) can act as endocrine disruptors, but few studies have investigated the effects of serum PFASs on estrogen homeostasis during pregnancy. The present study included 557 pregnant women in Tangshan City, North China, and determined 11 serum PFASs in the early term of pregnancy and three typical estrogens (estrone (E1), estradiol (E2) and estriol (E3)) in the early (n = 557), middle (n = 339), and late (n = 286) terms of pregnancy. Sociodemographic factors and diet information were obtained by structured questionnaires. After adjusting for potential confounders, multiple linear regression model demonstrated negative associations of natural logarithmic transformed serum perfluoroundecanoic acid (Ln PFUdA) with Ln E1and Ln E3 in the early term of pregnancy with ß coefficients of -0.060 (95% confidence interval (CI): -0.101 to -0.019) and -0.041 (95% CI: -0.070 to -0.011), respectively. Ln perfluorodecanoic acid (PFDA) was negatively associated with averaged E1 in the early and middle (EM) terms of pregnancy with a ß coefficient of -0.205 (95% CI: -0.357 to -0.053). Ln perfluorononanoic acid (PFNA) tended to be negatively associated with E2 in the late term of pregnancy with a ß coefficient of -0.134 (95% CI: -0.253 to -0.016) although p-value was slightly greater than 0.05 after false discovery rate (FDR) correction. Mixed effect model found that serum PFDA was negatively associated with E1 (ß = -0.123, 95% CI: -0.235 to -0.012) during the entire pregnancy. These findings suggested that exposure to PFASs disturbed estrogen homeostasis in pregnant women and the effects varied with the terms of pregnancy.

Exploring potential mechanisms of Suhexiang Pill against COVID-19 based on network pharmacology and molecular docking.

BACKGROUND: The traditional Chinese medicine prescription Suhexiang Pill (SHXP), a classic prescription for the treatment of plague, has been recommended in the 2019 Guideline for coronavirus disease 2019 (COVID-19) diagnosis and treatment of a severe type of COVID-19. However, the bioactive compounds and underlying mechanisms of SHXP for COVID-19 prevention and treatment have not yet been elucidated. This study investigates the mechanisms of SHXP in the treatment of COVID-19 based on network pharmacology and molecular docking. METHODS: First, the bioactive ingredients and corresponding target genes of the SHXP were screened from the traditional Chinese medicine systems pharmacology database and analysis platform database. Then, we compiled COVID-19 disease targets from the GeneCards gene database and literature search. Subsequently, we constructed the core compound-target network, the protein-protein interaction network of the intersection of compound targets and disease targets, the drug-core compound-hub gene-pathway network, module analysis, and hub gene search by the Cytoscape software. The Metascape database and R language software were applied to analyze gene ontology biological processes and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Finally, AutoDock software was used for molecular docking of hub genes and core compounds. RESULTS: A total of 326 compounds, 2450 target genes of SHXP, and 251 genes related to COVID-19 were collected, among which there were 6 hub genes of SHXP associated with the treatment of COVID-19, namely interleukin 6, interleukin 10, vascular endothelial growth factor A, signal transducer and activator of transcription 3 (STAT3), tumor necrosis factor (TNF), and epidermal growth factor. Functional enrichment analysis suggested that the effect of SHXP against COVID-19 is mediated by synergistic regulation of several biological signaling pathways, including Janus kinase/ STAT3, phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt), T cell receptor, TNF, Nuclear factor kappa-B, Toll-like receptor, interleukin 17, Chemokine, and hypoxia-inducible factor 1 signaling pathways. SHXP may play a vital role in the treatment of COVID-19 by suppressing the inflammatory storm, regulating immune function, and resisting viral invasion. Furthermore, the molecular docking results showed an excellent binding affinity between the core compounds and the hub genes. CONCLUSION: This study preliminarily predicted the potential therapeutic targets, signaling pathways, and molecular mechanisms of SHXP in the treatment of severe COVID-19, which include the moderate immune system, relieves the "cytokine storm," and anti-viral entry into cells.

Effects of histone H4 hyperacetylation on inhibiting MMP2 and MMP9 in human amniotic epithelial cells and in premature rupture of fetal membranes.

Histone modification is closely associated with several diseases. The aim of the current study was to investigate the associations among histone acetylation, matrix metalloproteinases (MMPs) and premature rupture of membranes (PROM) during pregnancy. A total of 180 puerperants were divided into three groups: i) Preterm-PROM (PPROM), ii) term-PROM (TPROM) and iii) full-term labor (FTL). Enzyme-linked immunosorbent assay (ELISA) kits and western blotting were used to determine the protein concentrations of MMP2, MMP9, histone deacetylase (HDAC)1, HDAC2 and HDAC6, and the protein levels of histone H4 lysine (H4K)5 and H4K8 acetylation, respectively, in three types of fetal membranes. Additionally, human amniotic epithelial cells were used to determine the effects of the HDAC inhibitors droxinostat and chidamide on cell viability, histone acetylation and the levels of MMP2, MMP9, HDAC1, HDAC2 and HDAC6 in vitro, using the Cell Counting Kit-8 assay, western blotting and ELISA, respectively. Furthermore, the effects of droxinostat and chidamide on the invasion and migration abilities of human amniotic epithelial cells were investigated using transwell assays. In fetal membranes, the activities of MMP2 and MMP9 increased in PPROM, but decreased in TPROM. Further, the expression of HDAC1 was decreased and histone hyperacetylation was increased in both PPROM and TRPOM. In vitro experiments revealed that 5 µM droxinostat and 0.5 µM chidamide selectively decreased the level of HDAC and induced acetylation of H4K5 and H4K8. Additionally, the aforementioned HDAC inhibitors reduced human amniotic epithelial cell viability, invasion and migration, and decreased the expression levels of MMP2 and MMP9. The current study revealed a high expression level of MMP2 and MMP9 in PPROM compared with TPROM and FL tissue, which was in accordance with previously published studies. Furthermore, the in vitro tests performed in the current study revealed the effect of histone H4 hyperacetylation on inhibiting MMP2 and MMP9 levels in vitro was similar to that observed in TPROM. The results obtained in the current study may be used as a theoretical guide for clinical treatment of premature rupture of membranes.

Serum perfluoroalkyl substances in relation to lipid metabolism in Chinese pregnant women.

Laboratory and epidemiologic studies suggested that exposure to perfluoroalkyl substances (PFASs) could affect lipid metabolisms, but data remain limited for pregnant women. A total of 436 pregnant women were selected in Tangshan City, North China. Serum levels of 11 PFASs were determined in the early term of pregnancy. Four lipids (total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL)) were measured in the late term of pregnancy. Of 11 PFASs, seven had a detection rate of greater than 70%. After adjusting for potential confounders, natural log-transformed perfluororohexanesulfonic acid (ln PFHxS) was positively associated with TC (ß: 0.184, 95% CI: 0.045-0.321), HDL (ß: 0.040, 95% CI: 0.001-0.083), and LDL (ß: 0.091, 95% CI: 0.001-0.185). Ln perfluoroundecanoic acid (PFUdA) was positively associated with HDL (ß: 0.021, 95% CI: 0.001-0.044), while Ln perfluorodecanoic acid (PFDA) was negatively associated with LDL (ß: -0.053, 95% CI: -0.098∼-0.009) and ln perfluorootanoic acid (PFOA) was negatively associated with LDL/HDL (ß: -0.042, 95% CI: -0.075∼-0.009). In principal component analysis, the component with a large loading of 31.3% for PFOA, perfluorononanoic acid (PFNA), PFDA and PFUdA showed a negative association with LDL/HDL. After serum concentrations of PFASs were categorized into quartiles, a higher level of TC was seen in the second quartile of PFOA or PFNA than the first quartile, but a lower LDL/HDL ratio was seen in the fourth quartile of PFOA, PFUdA or PFDA. These results suggested that exposure to PFASs has a potential to influence lipid metabolisms in pregnant women.

Serum Bisphenol A, glucose homeostasis, and gestational diabetes mellitus in Chinese pregnant women: a prospective study.

Lab studies have suggested that exposure to Bisphenol A (BPA) could disturb glucose homeostasis, but epidemiologic studies are limited and show inconsistent results for pregnant women. For this, 535 pregnant women were selected from a pregnant women cohort established in Tangshan City in North China between 2013 and 2014. Serum concentrations of BPA were measured in the early term of pregnancy, and fasting glucose and insulin levels were repeatedly measured in each of three terms of pregnancy (early, middle, and late). Gestational diabetes mellitus (GDM) were examined by Oral Glucose Tolerance Test (OGTT) in the middle and late terms of pregnancy. BPA was detected in 97.5% of pregnant women with a median of 6.50 ng/ml. Natural log-transformed BPA (Ln BPA) was positively associated with fasting glucose level (ß (95% CI): 0.038 (0.015~0.061)), fasting insulin level (0.195 (0.069~0.321)), and homeostasis model insulin resistance index (HOMA-IR) (0.226 (0.087~0.364)) in the middle term of pregnancy by multiple linear regression model after adjusting for potential confounders. After serum BPA levels were divided into three groups (low, middle, and high), BPA showed a positive dose-response relationship with blood glucose, insulin, and HOMA-IR in the middle term of pregnancy. Increased BPA concentration tended to increase the RR of GDM although not statistically significant (risk ratio: 2.51 (95% CI: 0.68~9.30) for high vs low tertile of BPA concentrations). These findings suggested that exposure to BPA might affect glucose homeostasis and the middle term of pregnancy was a potentially sensitive period.

Serum phytosterols associate with T helper 1 cytokine concentration in pregnant women.

The dietary phytosterols have been demonstrated to modulate CD4+ T-cell polarization in cells, animals, and humans. However, T helper (Th)1/Th2 dichotomy has rarely been correlated with phytosterols during pregnancy. The present study investigated associations between the serum cytokines and serum phytosterols in 100 pregnant women at 34- to 37-week gestation and their offspring. The results showed that serum concentrations of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and total Th1 cytokines were positively associated with serum ß-sitosterol level, adjusting for age, BMI, and serum cholesterol. Serum IFN-γ and total Th1 cytokine concentrations positively correlated with total phytosterol concentration, controlling age, BMI, and serum cholesterol. Moreover, none of the cytokines measured correlated with phytosterol concentration in the newborns. Our results show that serum Th1 cytokine concentrations, but not Th2 levels, are positively associated with serum phytosterols in pregnant women. These findings implicate that phytosterols modulate Th1/Th2 balance by inducing Th1 secretions in pregnant women.

PFOS, PFOA, estrogen homeostasis, and birth size in Chinese infants.

Laboratory studies have suggested that perfluoroalkyl substances (PFASs) could affect fetal growth by disrupting estrogen homeostasis, but there are limited data for human. For this, 424 mother-infant pairs were selected from a cohort established in Hebei Province of North China in 2013. Two typical PFASs, perfluorooctyl sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA), and three typical estrogens, estrone (E1), ß-estradiol (E2), and estriol (E3), were measured in cord serum. After adjusted for important covariates, serum PFOS was positively related to E1 and E3, but negatively related to E2. Serum PFOA was positively related to serum E1 and negatively related to head circumference at birth. Serum E2 was negatively related to head circumference, body weight, and body length at birth and serum E3 was positively related to body weight. Serum E3 mediated the relationship between serum PFOS and body weight. There were sex-specific differences for the associations between PFOS/PFOA and estrogens/birth size. These findings suggested that exposure to PFASs could affect estrogen homeostasis and fetal growth during pregnancy and that estrogens might mediate the association between exposure to PFASs and fetal growth.

Factors associated with exposure of pregnant women to perfluoroalkyl acids in North China and health risk assessment.

Perfluoroalkyl acids (PFAAs) have been frequently found in blood of pregnant women, but the predictors and potential health risk have not been well studied in China. We recruited 534 pregnant women in Tangshan City of Hebei Province in North China between 2013 and 2014 and measured five PFAAs in serum during their early term of pregnancy, including perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid (PFOS), perfluorohexane sulfonic acid (PFHxS), perfluoro­n­undecanoic acid (PFUdA), and perfluorononanoic acid (PFNA). We explored the factors associated with the levels of serum PFAAs and assessed associated health risks. Food consumption information was obtained by food frequency questionnaire covering 100 items. Multiple linear regression model was used to determine the associations of sociodemographic, anthropometric, and food factors with the concentrations of serum PFAAs. Some PFAAs in serum were positively associated with age and body mass index (BMI). Consumption of beans, aquatic products, and eggs was positively associated with the concentrations of several PFAAs after adjusting for important covariates. Pregnant women who ate more cereal, vegetables, mushrooms and alga tended to have lower levels of serum PFOA, PFOS and PFNA. The Hazard index (HI) for reproductive toxicity and developmental toxicity was below 0.8, and the HI for hepatotoxicity beyond 1 was found in 0.37% of pregnant women. These results suggested that age, BMI, and some food consumption were predictors for the exposure to PFAAs in Chinese pregnant women. More attention should be paid to the hepatotoxicity for these exposures.

Perfluoroalkyl substances, glucose homeostasis, and gestational diabetes mellitus in Chinese pregnant women: A repeat measurement-based prospective study.

BACKGROUND: Exposure to perfluoroalkyl substances (PFASs) can affect glucose homeostasis and has been suggested as a potential risk of diabetes mellitus, but data are limited for pregnant women. OBJECTIVES: We aimed to explore the associations of exposure to PFASs with glucose homeostasis and gestational diabetes mellitus (GDM) in Chinese pregnant women. METHODS: The current study was conducted in Hebei Province of Northern China between 2013 and 2014 and 560 pregnant women were recruited in their early term of pregnancy and two representative serum PFASs, perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS), were measured. In 385 pregnant women who completed oral glucose tolerance test (OGTT), the associations of serum PFOA and PFOS concentrations with fasting blood glucose (FBG), fasting insulin (FIns), and homeostasis model assessment of insulin resistance (HOMA-IR) in the early, middle, and late terms of pregnancy and occurrence of GDM were examined using linear and Cox proportional hazard regression models. The reproducibility of serum PFASs during pregnancy was assessed in 230 pregnant women. RESULTS: The intraclass correlation coefficients of serum PFASs, covariates, and outcomes based on averaged repeat measurement (0.35-0.96) were higher than those based on single measurement (0.16-0.92). Serum PFOA was positively associated with averaged FIns and HOMA-IR in the early, middle, and late terms of pregnancy and averaged blood glucose level at 1 h and 2 h of OGTT, but serum PFOS tended to be negatively associated with averaged FBG and OGTT blood glucose. The adjusted hazard ratios of GDM associated with serum PFOA and PFOS were 1.98 (95% confidence interval: 0.70-5.57; p-value: 0.197) and 0.71 (0.29-1.75; 0.453), respectively. CONCLUSIONS: Our data raised a possibility that exposure to PFASs might have different influences on glucose homeostasis and GDM in Chinese pregnant women. More lab and human studies are needed to further test the hypothesis and investigate potential mechanisms.

Molecular identification of Neofabraea species associated with bull's-eye rot on apple using rolling-circle amplification of partial EF-1α sequence.

A rolling-circle amplification (RCA) method with padlock probes targeted on EF-1α regions was developed for rapid detection of apple bull's-eye rot pathogens, including Neofabraea malicorticis, N. perennans, N. kienholzii, and N. vagabunda (synonym: N. alba). Four padlock probes (PLP-Nm, PLP-Np, PLP-Nk, and PLP-Nv) were designed and tested against 28 samples, including 22 BER pathogen cultures, 4 closely related species, and 2 unrelated species that may cause serious apple decays. The assay successfully identified all the bull's-eye rot pathogenic fungi at the level of species, while no cross-reaction was observed in all target species and no false-positive reaction was observed with all strains used for reference. This study showed that the use of padlock probes and the combination of probe signal amplification by RCA provided an effective and sensitive method for the rapid identification of Neofabraea spp. The method could therefore be a useful tool for monitoring bull's-eye rot pathogens in port quarantine and orchard epidemiological studies.

Noninvasive prenatal testing in routine clinical practice for a high-risk population: Experience from a center.

This study aimed to summarize the effects of noninvasive prenatal testing (NIPT) on aneuploidy among high-risk participants in Tangshan Maternal and Children Health Hospital.NIPT or invasive prenatal diagnosis was recommended to patients with a high risk of fetal aneuploidy from February 2013 to February 2014. Patients who exhibited eligibility and applied for NIPT from January 2012 to January 2013 were included in a comparison group. The rates of patients who underwent invasive testing, declined to undergo further testing, and manifested trisomies 21, 18, and 13 were compared between two groups. Follow-up data were obtained from the participants who underwent NIPT from 2013 to 2014.A total of 7223 patients (3018 and 4205 individuals before and after NIPT) were eligible for analysis. After NIPT was introduced in 2013 to 2014, 727 patients (17.3%) underwent invasive testing, 2828 preferred NIPT (67.3%), and 650 declined to undergo further testing (15.5%). A total of 34 cases of trisomies 21, 18, and 13 (0.8%) were found. In 2012 to 2013, 565 patients (18.7%) underwent invasive testing and 2453 declined to undergo further testing (81.3%). A total of 7 cases of trisomies 21, 18, and 13 were documented (0.2%). Of these cases, 24 were found from NIPT and 10 cases were found from invasive testing. The number of participants who declined to undergo further testing significantly decreased after NIPT was introduced (81.3% vs. 15.5%, P < 0.001). The sensitivity and specificity of NIPT for trisomies 21, 18, and 13 were 100% and 99.9%, respectively. The detection rates of NIPT for trisomies 21, 18, and 13 also significantly increased (0.2% vs. 0.8%, P < 0.001). By contrast, the overall rates of invasive testing remained unchanged (18.7% vs. 17.3%, P = 0.12). The positive predictive values of NIPT for trisomies 21, 18, and 13 were 100%, 83.3%, and 50.0%, respectively. The false positive rates of NIPT were 0% and 0.04%.With NIPT implementation in clinical practice, the rate of declining a follow-up test among high-risk women was decreased and the detection rate of prenatal chromosomal aneuploidy for trisomies 21, 18, and 13 was increased without requiring numerous invasive procedures.

Association Between Gene Polymorphisms on Chromosome 1 and Susceptibility to Pre-Eclampsia: An Updated Meta-Analysis.

BACKGROUND This meta-analysis enabled us to obtain a precise estimation of the association between gene polymorphisms on chromosome 1 (MTHFR, AGT, F5, IL-10, LEPR) and the susceptibility to pre-eclampsia (PE) in order to reach a uniform conclusion. MATERIAL AND METHODS Web of Science, PubMed, EMBASE, Cochran Library (CENTRAL), and Chinese databases (Chinese National Knowledge Infrastructure-CNKI and Wan Fang) were electronically searched to select relevant studies for this meta-analysis. We selected 95 case-control studies investigating 5 genes (MTHFR, AGT, F5, IL-10, and LEPR) with 8 SNPs. Odds ratios (OR) with their 95% confidence intervals (CI) were used for estimating the association. RESULTS A total of 16 646 PE patients and 28 901 normal-pregnancy patients were included in this meta-analysis. The overall results suggested that rs1801133 of MTHFR (OR=1.17, 95% CI: 1.05-1.13) and rs6025 of F5 (OR=1.53, 95%CI: 1.07-2.20) are significantly associated with PE, whereas rs1801131 of MTHFR, rs699 and rs4762 of AGT, rs1800896 and rs1800871 of IL-10, and rs1137101 of LEPR have no significant association with PE. Subgroup analysis by ethnicity revealed that, except for MTHFR rs1801133 and F5 rs6025 in Caucasians, which were significantly associated with an increased risk of PE, none of these SNPs were significantly associated with PE. As suggested by a symmetric funnel plot in conjunction with the Egger's test, there was no significant publication bias in MTHFR rs1801133 (P=0.318) and rs1801131 (P=0.204), F5 rs6025 (P=0.511), LEPR rs1137101 (P=0.511), AGT rs4762 (P=0.215) and rs699 (P=0.482), IL-10 rs1800871 (P=0.955), and rs1800896 (P=0.144). CONCLUSIONS This meta-analysis provides evidence that MTHFR rs1801133 and F5 rs6025 are associated with an increased risk of PE, especially in Caucasians. However, we do not have sufficient evidence to conclude there is a significant association between other gene polymorphisms and PE.

Non-thermal plasma treatment of Radix aconiti wastewater generated by traditional Chinese medicine processing.

The wastewater effluent from Radix aconiti processing, an important step in the production processes of traditional Chinese medicine (TCM), is a type of toxic wastewater and difficult to treat. Plasma oxidation methods have emerged as feasible techniques for effective decomposition of toxic organic pollutants. This study examined the performance of a plasma reactor operated in a dielectric barrier discharge (DBD) to degrade the effluent from R. aconiti processing. The effects of treatment time, discharge voltage, initial pH value and the feeding gas for the reactor on the degradation of this TCM wastewater were investigated. A bacterium bioluminescence assay was adopted in this study to test the toxicity of the TCM wastewater after non-thermal plasma treatment. The degradation ratio of the main toxic component was 87.77% after 60min treatment with oxygen used as feed gas and it was 99.59% when the initial pH value was 8.0. High discharge voltage and alkaline solution environment were beneficial for improving the degradation ratio. The treatment process was found to be capable of reducing the toxicity of the wastewater to a low level or even render it non-toxic. These experimental results suggested that the DBD plasma method may be a competitive technology for primary decomposition of biologically undegradable toxic organic pollutants in TCM wastewater.

Dickeyafangzhongdai sp. nov., a plant-pathogenic bacterium isolated from pear trees (Pyrus pyrifolia).

Gram-stain-negative, pectinolytic bacteria were repeatedly isolated from pear trees displaying symptoms of bleeding canker in China. Three strains, JS5T, LN1 and QZH3, had identical 16S rRNA gene sequences that shared 99 % similarity to the type strain of Dickeya dadantii. Phylogenetic analysis of strains JS5T, LN1 and QZH3 with isolates representing all species of the genus Dickeya and related Pectobacterium species supported their affiliation to Dickeya. Multi-locus sequence typing employing concatenated sequences encoding recA, fusA, gapA, purA, rplB, dnaX and the intergenic spacer illustrated a phylogeny which placed strains JS5T, LN1 and QZH3 as a distinct clade, separate from all other species of the genus Dickeya. Average nucleotide identity values obtained in comparison with all species of the genus Dickeya supported the distinctiveness of strain JS5T within the genus Dickeya. Additionally, all three strains were phenotypically distinguished from other species of the genus Dickeya by failing to hydrolyse casein, and by producing acids from (-)-d-arabinose, (+)melibiose, (+)raffinose, mannitol and myo-inositol, but not from 5-keto-d-gluconate or ß-gentiobiose. The name Dickeya fangzhongdai sp. nov. is proposed to accommodate these strains; the type strain is JS5T (=CGMCC 1.15464T=DSM 101947T).

Genomic sequence, organization and characteristics of a new nucleopolyhedrovirus isolated from Clanis bilineata larva.

BACKGROUND: Baculoviruses are well known for their potential as biological agents for controlling agricultural and forest pests. They are also widely used as expression vectors in molecular cloning studies. The genome sequences of 48 baculoviruses are currently available in NCBI databases. As the number of sequenced viral genomes increases, it is important for the authors to present sufficiently detailed analyses and annotations to advance understanding of them. In this study, the complete genome of Clanis bilineata nucleopolyhedrovirus (ClbiNPV) has been sequenced and analyzed in order to understand this virus better. RESULTS: The genome of ClbiNPV contains 135,454 base pairs (bp) with a G+C content of 37%, and 139 putative open reading frames (ORFs) of at least 150 nucleotides. One hundred and twenty-six of these ORFs have homologues with other baculovirus genes while the other 13 are unique to ClbiNPV. The 30 baculovirus core genes are all present in ClbiNPV. Phylogenetic analysis based on the combined pif-2 and lef-8 sequences places ClbiNPV in the Group II Alphabaculoviruses. This result is consistent with the absence of gp64 from the ClbiNPV genome and the presence instead of a fusion protein gene, characteristic of Group II. Blast searches revealed that ClbiNPV encodes a photolyase-like gene sequence, which has a 1-bp deletion when compared with photolyases of other baculoviruses. This deletion disrupts the sequence into two small photolyase ORFs, designated Clbiphr-1 and Clbiphr-2, which correspond to the CPD-DNA photolyase and FAD-binding domains of photolyases, respectively. CONCLUSION: ClbiNPV belongs to the Group II Alphabaculoviruses and is most closely related to OrleNPV, LdMNPV, TnSNPV, EcobNPV and ChchNPV. It contains a variant DNA photolyase gene, which only exists in ChchNPV, TnSNPV and SpltGV among the baculoviruses.

Identification of a single-nucleocapsid baculovirus isolated from Clanis bilineata tsingtauica (Lepidoptera: Sphingidae).

A nucleopolyhedrovirus isolated from infected larvae of Clanis bilineata tsingtauica was characterized. Electron microscopical studies on the ultrastructure of C. bilineata nucleopolyhedrovirus (ClbiSNPV) occlusion bodies (OBs) showed several virions (up to 16) with a single nucleocapsid packaged within a single viral envelope. The diameter of the OBs was 0.77-1.7 mum with a mean of 1.13 +/- 0.19 mum. The complete sequence of the ClbiSNPV polyhedrin (polh) gene contained 741 nucleotides, predicting a protein of 246 amino acids. Phylogenetic analyses using the complete sequence of the polh genes indicated that ClbiSNPV clusters with Group II NPVs. This is the first record of a baculovirus from C. bilineata.

[Extraction of mitochondrial DNA of Tilletia indica Mitra and application of the ATP6 gene on fungal phylogenetic analysis].

Tilletia indica Mitra, an important pest around the world which is the causative agent of Karnal Bunt of wheat, was very close to Tilletia walkeri phylogenetically and morphologically. 69.6 ng mitochondrial DNA (mtDNA) of Tilletia indica obtained from 6.4 mg total DNA by the method of CsCl/bisbenzimide density gradient ultracentrifugation can be used for electrophoresis, cloning, enzyme restriction analysis and PCR amplification. The ATP6 gene sequence was cloned from the fragment of mtDNA and sequenced for analysis of phylogenetic tree with the other related sequences in GenBank by the software PAUP to reveal the phylogenetic relationships within the Tilletia species.