Climacocystaceae fam. nov. and Gloeoporellaceae fam. nov., two new families of Polyporales (Basidiomycota).

Polyporales is a diverse group of Agaricomycetes including more than 2,500 species belonging to 255 genera and 18 families. Recently, many studies focused on the classification of Polyporales, but the familial placements of some taxa remain uncertain. In this study, two new families, Climacocystaceae and Gloeoporellaceae of Polyporales, are proposed based on morphological characters and molecular data. Phylogenetic analyses of the two new families are inferred from the DNA sequences of the internal transcribed spacer regions (ITS), the large subunit of nuclear ribosomal RNA gene (nLSU), the largest subunit of RNA polymerase II gene (RPB1), the second largest subunit of RNA polymerase II gene (RPB2), and the translation elongation factor 1-α gene (TEF1). Furthermore, the divergence time of Polyporales was estimated as an additional taxonomic criterion based on the conserved regions of five DNA fragments (5.8S, nLSU, RPB1, RPB2, and TEF1). Bayesian evolutionary analysis revealed that the ancestor of Polyporales splits with a mean stem age of 136.53 Mya with a 95% highest posterior density (HPD) of 118.08-158.06 Mya. The mean stem ages of the families within Polyporales originated between 66.02 and 119.22 Mya, of which Climacocystaceae occurred in a mean stem age of 77.49 Mya with a 95% HPD of 61.45-93.16 Mya, and Gloeoporellaceae occurred in a mean stem age of 88.06 Mya with a 95% HPD of 67.15-107.76 Mya.

Four new species of Agaricus subgenus Spissicaules from China.

Agaricus subgenus Spissicaules is widely distributed in the world. In this study, 114 specimens were included in multigene phylogenetic analyses that allowed a better circumscription of the four sections in A. subg. Spissicaules. Three new species from China, A. brunneovariabilis, A. beijingensis, and A. planiceps, are described and placed in different sections. The invalidly described A. catenariocystidiosus is validated here as an additional new species. Comparisons between the sequences of the three closely related species A. thiersii, A. parasubrutilescens, and A. linzhiensis support the distinction between these three species despite the paraphyly of A. linzhiensis, which results from its high intraspecific variability with numerous heteromorphisms.

Time spent outside of the hospital, CKD progression, and mortality: a prospective cohort study.

PURPOSE: Home time-being out of any healthcare facility-has been proposed as a patient-centered outcome. This novel measure has not been investigated in patients with chronic kidney disease (CKD). The aim of this study is to determine whether there was an association between home time and occurrence of end-stage renal disease (ESRD) or all-cause mortality during 1 year of follow-up. METHODS: We assembled a prospective cohort of patients with CKD not requiring dialysis at the Nephrology Center of First Affiliated Hospital of Jiaxing University between May 2014 and April 2017 and followed up for 1 year. Home time was calculated as the number of days spent out of a hospital, rehabilitation facility, or skilled nursing facility. Outcomes included progression to ESRD and all-cause mortality. RESULTS: Among 943 patients, 882 (93.5%) had complete follow-up through 1 year. Mean home time was 246.9 ± 126.7 days. In regression analysis, several patient characteristics were associated with significantly reduced home time, including diabetes mellitus, cardiovascular disease, and albuminuria. Home time was strongly correlated with time-to-event endpoints of ESRD (τ=0.324) and all-cause mortality (τ=0.785). CONCLUSIONS: Home time is significantly reduced for patients with CKD not requiring dialysis and is highly correlated with traditional time-to-event endpoints. Home time serves as a novel, easily calculated, patient-centered outcome that may reflect effect of interventions on future CKD research.

Patterns of PCR Amplification Artifacts of the Fungal Barcode Marker in a Hybrid Mushroom.

The polymerase chain reaction (PCR) is widely used in modern biology and medicine. However, PCR artifacts can complicate the interpretation of PCR-based results. The internal transcribed spacer (ITS) region of the ribosomal RNA gene cluster is the consensus fungal barcode marker and suspected PCR artifacts have been reported in many studies, especially for the analyses of environmental fungal samples. At present, the patterns of PCR artifacts in the whole fungal ITS region (ITS1+5.8S+ITS2) are not known. In this study, we analyzed the error rates of PCR at three template complexity levels using the divergent copies of ITS from the mushroom Agaricus subrufescens. Our results showed that PCR using the Phusion® High-Fidelity DNA Polymerase has a per nucleotide error rate of about 4 × 10-6 per replication. Among the detected mutations, transitions were much more frequent than transversions, insertions, and deletions. When divergent alleles were mixed as templates in the same reaction, a significant proportion (∼30%) of recombinant molecules were detected. The in vitro mixed-template results were comparable to those obtained from using the genomic DNA of the original mushroom specimen as template. Our results indicate that caution should be in place when interpreting ITS sequences from individual fungal specimens, especially those containing divergent ITS copies. Similar results could also happen to PCR-based analyses of other multicopy DNA fragments as well as single-copy DNA sequences with divergent alleles in diploid organisms.

Species Diversity, Phylogeny, Divergence Time, and Biogeography of the Genus Sanghuangporus (Basidiomycota).

"Sanghuang" is a popular fungus used as a Chinese traditional medicine. In fact, it represents a group of fungi belonging to the genus Sanghuangporus, but little is known about its origin and biogeography. The aim of this study was to characterize the molecular relationships, origin and biogeographical distribution of Sanghuangporus. The multi-locus phylogenetic analyses were used to infer the phylogenetic relationships. In addition, based on Bayesian evolutionary analysis using sequences from the internal transcribed spacer (ITS), nuclear large subunit rDNA (nLSU), translation elongation factor 1-α (EF1-α), and the largest and second largest subunits of RNA polymerase II (RPB1 and RPB2), we used a fungus fossil-based approach to gain insight into the divergence time of species in Sanghuangporus. The molecular phylogeny strongly supports the monophyly of Sanghuangporus (MP = 100%, ML = 100%, and BPP = 1.00), and 13 species are recognized in this genus. The Bayesian uncorrelated lognormal relaxed molecular clock using BEAST and reconstructed ancestral areas indicate that the maximum crown age of Sanghuangporus is approximately 30.85 million years. East Asia is the likely ancestral area (38%). Dispersal and differentiation to other continents then occurred during the late Middle Miocene and Pliocene. The ancestor of Sanghuangporus probably originated in palaeotropical Northeast Asia and covered Northeast Asia and East Africa during the Oligocene-Miocene, hosted by plants that expanded via the "Gomphotherium Landbridge." Six kinds of dispersal routes are proposed, including intercontinental dispersal events of three clades between Northeast Asia and East Africa, between East Asia and North America, and between Northeast Asia and Europe.

Bioaccumulation, tissue distribution and joint toxicity of erythromycin and cadmium in Chinese mitten crab (Eriocheir sinensis).

The bioaccumulation of erythromycin (ETM) and cadmium (Cd) in Chinese mitten crab (Eriocheir sinensis) and subsequent toxicity on pathological changes and enzymatic activities were investigated during 21-day exposure to ETM, Cd, and Cd + ETM mixture. The bioaccumulation of Cd and ETM residues in crab tissues decreased as gill > hepatopancreas > muscle > ovary, with higher Cd bioaccumulation than ETM. The highest Cd bioaccumulation in crab reached 1.15 mg/g dry weight in gill and 461.29 µg/g in hepatopancreas, on the 14th day of Cd treatment. Cd exposure promoted the bioaccumulation of ETM in four tissues. ETM exposure caused tubular vacuolization in epithelial and edema and degeneration of hepatic ducts in hepatopancreas, and disconnected gill epithelial layer and indistinctly cellular structure in gill. During Cd exposure, mitochondria acted as a main biomarker to identify the damage, including reduced and swollen mitochondria, and broken mitochondrial structure. Moreover, Chinese mitten crab showed defence capability against ETM and Cd exposure by physiological adjustment of metabolic enzymes glutathione S-transferase activity.

Low serum levels of vitamin D are associated with anxiety in children and adolescents with dialysis.

Anxiety is a frequent and serious complication of children and adolescents receiving dialysis. Low serum vitamin D levels have been associated with anxiety in non-pediatric patients. This study sought to examine the possible association between serum vitamin D levels and the presence of anxiety in children and adolescents with dialysis in China. A total of 156 pediatric patients who were on hemodialysis or peritoneal dialysis and 100 healthy controls were included in the current study. Serum 25-hydroxyvitamin D [25(OH)D] levels were measured by using a competitive protein-binding assay. Anxiety was assessed by using the Chinese version of the Screen for Child Anxiety Related Emotional Disorders (SCARED, = 25 as cutoff). Among 156 patients, 110 had a current anxiety (70.5%) and 46 did not (29.5%). Serum levels of 25(OH)D were significantly lower in patients with anxiety than in normal controls (19.4 ± 10.3 vs. 38.6 ± 15.5 ng/ml, P < 0.001). Serum 25(OH)D levels (≤15.0 ng/ml) were independently associated with the existent of anxiety in children and adolescents receiving dialysis (OR 4.650, 95% CI 1.663-13.001, P = 0.003). Our research demonstrates that low serum levels of vitamin D are independently associated with anxiety among children and adolescents on dialysis, which needs to be confirmed in future experimental and clinical studies.

Tropic origins, a dispersal model for saprotrophic mushrooms in Agaricus section Minores with descriptions of sixteen new species.

Agaricus section Minores contains the richest species diversity within the genus. Its Phylogeny is firstly presented by a Maximum Likelihood tree generated through DNA sequences from four gene regions of 91 species. Furthermore, a molecular dating analysis is conducted used those sequences, and it provided the divergence times of the clades within section Minores. Study showed section Minores has a tropical origin. Four main dispersal routes are proposed: (1) species from South Asia migrated through the Tibetan Plateau and reached Europe ca. 9-13 Ma; (2) species from out of South Asia dispersed to Europe in the earlier time of ca. 22 Ma; (3) species from South Asia dispersed through North Asia to Alaska, and reached West America around ca. 9 Ma; and (4) species from South Asia dispersed south and reached Oceania by at least three invading events about ca. 9, 12 and 16-18 Ma respectively. Those routes excepting the second route coincide with those of ectomycorrhizal mushrooms. To know whether the second route existed in the saprotrophic mushrooms requires further studies, and the fourth route may explain why the secotioid species occurring in Australia are morphologically similar but cluster in different phylogenetic clades. This study also demonstrates a great biodiversity of A. section Minores in China. Sixteen new species and three new records are introduced from China with morphological descriptions, illustrations, color photographs and phylogenetic analyses.

Bioaccumulation of sulfadiazine and subsequent enzymatic activities in Chinese mitten crab (Eriocheir sinensis).

The bioaccumulation of sulfadiazine and subsequent enzymatic activities in Chinese mitten crab (Eriocheir sinensis) were studied in microcosms, by exposing to 50, 100, 500 and 1000ng/L of sulfadiazine for 44days. An effective method for extracting sulfadiazine in crab tissues was established by modifying the cleanup method after ultrasound extraction, with improved recoveries of 61.8%, 93.7% and 100.5% in gill, muscle and liver samples. The results showed that sulfadiazine residues were all <3ng/g dry weight in different tissues, and that sulfadiazine bioaccumulation in crab was not dose-dependent. A significantly negative correlation was observed between acetylcholinesterase activity and the residue concentration of sulfadiazine during exposure to 50ng/L and 1000ng/L, and between alkaline phosphatase and sulfadiazine residues in the 100ng/L exposure group in the gill, suggesting that the two enzymes played an important role in the metabolism of sulfadiazine in crab.

Effects of ibuprofen, diclofenac and paracetamol on hatch and motor behavior in developing zebrafish (Danio rerio).

Non-steroidal anti-inflammatory drugs (NSAIDs) which are widely used as pain relief medicines are causing increasing environmental concern due to their incomplete removal in wastewater treatment plant and potential toxicity on endocrine, kidney and reproduction in teleost fish. This study focused on the effects of widely used ibuprofen, diclofenac and paracetamol on the hatch and motor ability of early-stage zebrafish, by exposing embryos to the target chemicals at 5, 50 and 500 µg/L starting from 6 h postfertilization (hpf). A significant reduction in hatch rate at 55 hpf was caused by both ibuprofen (-63%) and diclofenac (-58%) at 500 µg/L. Exposure to high concentration of ibuprofen significantly decreased the spontaneous movement by 25%, and reduced the free swimming distance, duration and speed under dark condition by 41%, 29% and 30%, respectively. High concentration of diclofenac also caused 23% decrease in spontaneous movement, and reduced the swimming distance as well as active duration by 17% and 13% under light stimulation. In comparison, the exposure to paracetamol did not cause any notable effect. Among neuron related genes tested, the expression of neurog1 was down-regulated from ibuprofen and diclofenac exposure by 19% and 26%, while the expression of neurod1 was up-regulated only by ibuprofen (31%). These findings indicated that ibuprofen and diclofenac significantly affected embryo locomotivity and were potentially neurotoxic, thus posing threats to zebrafish development.

[Fluorescence Characterization of Fractionated Colloids in Wastewaters Received by Huangpu River].

The spectral characteristics of fractionated colloids in wastewaters from Huangpu River were isolated and investigated via cross flow ultrafiltration systems with different molecular mass of membranes and three-dimensional excitation-emission matrix fluorescence spectroscopy. The results showed that the colloidal concentration (CC) and colloidal organic carbon content (COC) of all size fractions in all samples were in the ranges 9.60-32.50 mg·L-1 and 0.03-6.25 mg·L-1, respectively. Moreover, both CC and COC values increased as the colloidal size increased. The protein- (i.e., peaks D and T) and humic-like (peaks C and A) materials were present in all the size fractions (except for the Mr 300×103-1 µm size fraction of the influent sample), of which protein- and humic-like materials were mainly in the Mr 1×103-5×103 and Mr 10×103-300×103 size fractions, respectively. In addition, fluorescence indices (including the fluorescence index (FI), humification index (HIX), and the index of recent autochthonous contribution (BIX)) were introduced to determine the source and spectral characteristic of the selected colloids. These indices indicated that the relatively small colloids with high aromaticity and hydrophobicity mainly originated from terrestrial sources, whereas the relatively large colloids were predominantly from the recent autochthonous organic matter and the protein-like components.

Phylogeny and taxonomy of Favolus (Basidiomycota).

Favolus is a natural genus that is closely related to Polyporus. A taxonomic study of Favolus was carried out based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2 = ITS) and 18S and 28S genes of the nuc rDNA, translation elongation factor 1-α gene (TEF1-α), the mt 16S rDNA gene (mt rrnS), ß-tubulin gene (TBB1), and the RNA polymerase II largest subunit (RPB1) and second largest subunit (RPB2) genes. Fourteen specimens collected from subtropical to tropical regions in China were shown to be different from existing Favolus species. They are described and illustrated here as four new species, namely, Favolus niveus, F. septatus, F. pseudoemerici, and F. subtropicus.

Taxonomy and Phylogeny of Polyporus Group Melanopus (Polyporales, Basidiomycota) from China.

Melanopus is a morphological group of Polyporus which contains species with a black cuticle on the stipe. In this article, taxonomic and phylogenetic studies on Melanopus group were carried out on the basis of morphological characters and phylogenetic evidence of DNA sequences of multiple loci including the internal transcribed spacer (ITS) regions, the large subunit nuclear ribosomal RNA gene (nLSU), the small subunit nuclear ribosomal RNA gene (nSSU), the small subunit mitochondrial rRNA gene sequences (mtSSU), the translation elongation factor 1-α gene (EF1-α), the largest subunit of RNA polymerase II (RPB1), the second largest subunit of RNA polymerase II (RPB2), and ß-tubulin gene sequences (ß-tubulin). The phylogenetic result confirmed that the previously so-called Melanopus group is not a monophyletic assemblage, and species in this group distribute into two distinct clades: the Picipes clade and the Squamosus clade. Four new species of Picipes are described, and nine new combinations are proposed. A key to species of Picipes is provided.

Transcriptional and morphological effects of tamoxifen on the early development of zebrafish (Danio rerio).

Tamoxifen is a widely used anticancer drug with both an estrogen agonist and antagonist effect. This study focused on its endocrine disrupting effect, and overall environmental significance. Zebrafish embryos were exposed to different concentrations (0.5, 5, 50 and 500 µg l(-1) ) of tamoxifen for 96 h. The results showed a complex effect of tamoxifen on zebrafish embryo development. For the 500 µg l(-1) exposure group, the heart rate was decreased by 20% and mild defects in caudal fin and skin were observed. Expressions of a series of genes related to endocrine and morphological changes were subsequently tested through quantitative real-time polymerase chain reaction. Bisphenol A as a known estrogen was also tested as an endocrine-related comparison. Among the expression of endocrine-related genes, esr1, ar, cyp19a1b, hsd3b1 and ugt1a1 were all increased by tamoxifen exposure, similar to bisphenol A. The cyp19a1b is a key gene that controls estrogen synthesis. Exposure to 0.5, 5, 50 and 500 µg l(-1) of tamoxifen caused upregulation of cyp19a1b expression to 152%, 568%, 953% and 2024% compared to controls, higher than the effects from the same concentrations of bisphenol A treatment, yet vtg1 was suppressed by 24% from exposure to 500 µg l(-1) tamoxifen. The expression of metabolic-related genes such as cyp1a, cyp1c2, cyp3a65, gpx1a, gstp1, gsr and genes related to observed morphological changes such as krt17 were also found to be upregulated by high concentrations of tamoxifen. These findings indicated the potential environmental effect of tamoxifen on teleost early development. Copyright © 2015 John Wiley & Sons, Ltd.

New species of Cystolepiota from China.

In this paper, a new species, Cystolepiota pseudofumosifolia, is introduced. C. pseudofumosifolia is characterized by granulose or powdery pileus with an anatomic structure that is loosely globose, as well as ellipsoid cells in chains in the pileus covering the cheilocystidia. This new species is compared to the related and similar Cystolepiota species in morphology and molecular phylogeny based on Internal transcribed spacer sequences. Both types of data support our specimens as a new species in the genus Cystolepiota.

Tidal impact on the dynamic behavior of dissolved pharmaceuticals in the Yangtze Estuary, China.

The dynamic behavior of 24 dissolved pharmaceuticals over tidal cycles in the Yangtze Estuary, China was studied to assess the tidal impact on the fate of pharmaceutical residues. The results show that most pharmaceuticals were frequently detected with concentrations from below detection to 27.2 ng/L, with sulfamethoxazole, sulfamethazine, erythromycin, thiamphenicol and florfenicol dominating. During tidal cycles, pharmaceutical concentrations decreased during tidal rise, then increasing during tidal receding for all locations, except at site S2 which showed an opposite trend due to unique water movement there. It was observed that most compounds showed a non-conservative behavior, while diazepam and sulfamethoxazole displayed a conservative behavior. The pharmaceutical concentrations were found to increase with dissolved organic carbon (DOC) concentration, suggesting DOC as a carrier of pharmaceuticals. In addition, many compounds showed a significant negative relationship with suspended particulate matter (SPM) concentration, indicating SPM-water interactions as a control of pharmaceutical behavior in estuarine environment.

[Concentration and distribution characteristics of estrogen in aquatic organism from Chongming Island].

Five typical estrogens, including estrone (E1), 17ß-estradiol (E2), estriol (E3), 17α-ethynyl estradiol (EE2) and bisphenol A (BPA), were simultaneously determined in typical aquatic organisms from Chongming Island by ultrahigh-performance liquid chromatography equipped with tandem mass spectrometry (UHPLC-MS2). The distribution pattern of estrogens was studied and preliminary heath risk assessment was discussed. The results showed that the total dry weight concentrations of estrogens were from 1.1 ng x g(-1) to 7.38 ng x g(-1) with a mean of 4.25 ng x g(-1). The lipid content-standardized estrogens concentrations were in the range of 5.01- 83.41 ng x g(-1) with a mean of 40.75 ng x g(-1), showing a descending order of fish > shrimp > crab. Fish and crab, fish and shrimp showed a significant difference in lipid content-standardized estrogen concentrations, whereas no significant difference was found between crab and shrimp. Compared to BPA, the dry weight concentrations of steroids (E1, E2, E3 and EE2) were found at lower levels (0.17-0.69 ng x g(-1)) with lower detection frequency (< 66.67%) in aquatic organisms from Chongming Island. In contrast, the detection rate of BPA reached 100%, and the average dry weight concentration detected was 2.60 ng x g(-1). The results of health risk assessment showed the concentration of estrogens in aquatic products from Chongming Island could not cause any adverse population-level effects.

Simultaneous determination of steroidal and phenolic endocrine disrupting chemicals in fish by ultra-high-performance liquid chromatography-mass spectrometry/mass spectrometry.

A sensitive and reliable analytical method based on pressurized liquid extraction (PLE) and ultra-high-performance liquid chromatography equipped with tandem mass spectrometry (UHPLC-MS/MS) has been developed for simultaneously determining the steroidal and phenolic endocrine disrupting chemicals (EDCs) in fish. The most effective extraction of the target EDCs is achieved by using PLE with on-line purification and the parameters have been optimized as follows: extraction solvent - methanol-acetonitrile (1:1, v/v), on-line purification material - 5 g alumina (5% water), extraction - 3 cycles, static extraction time - 5 min and extraction temperature - 60°C. Compared to the Oasis hydrophilic-lipophilic balance (HLB) solid phase extraction (SPE), freezing-lipid filtration combined with n-hexane defatting clean-up obtains much better recoveries of the target compounds and provide cleaner extracts. The matrix effect (ME) is generally eliminated by using an internal standard method. At spiking levels of 5, 50, and 100 ng/g, the mean recoveries vary from 71.2% to 108% for the target EDCs with a relative standard deviation (RSD) less than 16%. The method limit of detection (LOD) and limit of quantitation (LOQ) are 0.04-0.08 ng/gdw and 0.07-0.27 ng/gdw, respectively. The established method has been successfully applied to fish samples from the local market to determine the target EDCs.

Matrix effect in high-performance liquid chromatography-tandem mass spectrometry analysis of antibiotics in environmental water samples.

This paper describes the matrix effect during the analysis of ten antibiotic compounds in water by SPE followed by HPLC-ESI-MS/MS. The target analytes were tetracycline, oxytetracycline (tetracyclines), sulfathiazole, sulfamethazine, sulfadiazine (sulfonamides), erythromycin-H(2)O, roxithromycine, spiramycin (macrolides), ofloxacin, and norfloxacin (quinolones). The matrix effect was examined for internal standards and the target analytes in five different water matrixes, with signal suppression being increased in the order: ultrapure water, tap water, river water, sewage effluent, and sewage influent. A combined application of the internal standards and matrix-matched extract calibration was shown to be successful in compensating the matrix effect for the analytes. The procedural recovery of the target compounds in sewage effluents and influents was higher than in river water samples, which was further enhanced by sample acidification to pH 2. The validity of the internal standard based matrix-matched calibration approach was verified by the standard addition method.

Spry2 expression correlates with BRAF mutation in thyroid cancer.

BACKGROUND: BRAF mutations activate the mitogen-activated protein kinase pathway and often confer an aggressive thyroid cancer (TC) phenotype. Spry2 is an inducible negative feedback regulator of the mitogen-activated protein kinase (MAPK) pathway. The aim of this study was to investigate the role of Spry2 in TC. METHODS: TC cell lines were analyzed for Spry2 expression and MAPK pathway activation. Cells were treated with MEK inhibitor and Spry2 small hairpin RNA. Cells were analyzed for Spry2 expression and MEK/ERK phosphorylation (pMEK, pERK). Thirty human papillary TCs were analyzed for mitogen-activated protein kinase pathway activating mutations and Spry2 expression. RESULTS: Increased baseline pMEK levels and Spry2 expression was found in BRAF V600E mutant (BRAF+) cells. MEK inhibition in BRAF+ cells showed decreased Spry2 expression and decreased pMEK/pERK levels. From our tissue samples, 10 papillary TCs had BRAF mutation, and increased Spry2 expression was found only in BRAF+ tumors. CONCLUSION: Spry2 expression correlates with BRAF status in vitro and in human tissue. Spry2 may serve as a negative feedback regulator of the mitogen-activated protein kinase pathway in BRAF+ TC. Increased Spry2 expression may serve as a surrogate marker of mitogen-activated protein kinase pathway activation with prognostic and therapeutic implications.