Incomplete radiofrequency ablation following transarterial chemoembolization accelerates the progression of large hepatocellular carcinoma.

Purpose: To examine post-operative progression and risk impact of insufficient radiofrequency ablation (RFA) following transarterial chemoembolization (TACE) for the prognosis of large hepatocellular carcinoma (HCC). Materials and Methods: From January 2014 to January 2021 were analyzed. A total of 343 patients with large HCC (diameter >5 cm) who received TACE combined with RFA were enrolled and were divided into two groups: complete ablation (CA, n = 172) and insufficient ablation (IA, n = 171). Overall survival (OS) and progression-free survival (PFS) were determined by the Kaplan-Meier curve and compared with the log-rank test. To find parameters influencing OS and PFS, clinicopathological variables underwent univariate and multivariate analysis. Results: The cumulative 1-, 3-, and 5-year OS and PFS rates of the CA group were significantly higher than that of the IA group (P < 0.001). 25 (41%) patients in local tumor progression (LTP), 36 (59%) in intrahepatic distant recurrence (IDR), and 0 (0%) in extrahepatic distant recurrence (EDR) in the CA group. 51 (32.1%) patients in LTP, 96 (60.4%) patients in IDR, and 12 (7.5%) cases in EDR in the IA group. The recurrence patterns of the two groups were statistically significant difference (P = 0.039). In multivariate analysis, inadequate ablation and conjunction with TKIs were both significant risk factors for OS and PFS. Apart from these, older age and >7 cm of tumor size were indicators of poor OS and multiple tumors were indicators of poor PFS. Conclusion: Insufficient ablation causes a poor survival outcome of TACE combined with RFA for large HCC, particularly, which can promote IDR.

Anal fistulotomy with one-stage shaped skin grafting for intersphincter anal fistulas: study protocol on a multicentre randomised controlled trial.

BACKGROUND: Anal fistulas are mainly treated via surgery. They can be difficult to treat without surgical intervention. Numerous procedures, such as fistulectomy and fistulotomy, are performed to treat anal fistulas and achieve good effects. However, the wounds created through fistulectomy and fistulotomy take a long time to heal. Therefore, a multicentre randomised controlled trial (RCT) is proposed to study the efficacy of one-stage shaped skin grafting at the surgical wound to heal low simple intersphincter anal fistulas. METHODS: This study is a multicentre, hospital-based RCT. It will be performed at three hospitals. A total of 104 patients with low simple intersphincter anal fistulas who meet the inclusion criteria will be included in this trial and will be allocated randomly to two groups (test and control groups). The patients in the test group will receive one-stage anal fistulotomy surgery combined with shaped skin grafting, and those in the control group will undergo anal fistulotomy only. All the operations will be performed by attending colorectal surgeons or surgeons of a higher level. Effectiveness and safety indicators will be observed, recorded and analysed. DISCUSSION: Anal fistulotomy can heal low simple intersphincter anal fistulas effectively and safely with a low recurrence rate. Skin grafts promote wound epithelisation significantly. We believe that skin grafting can treat low simple intersphincter fistulas with a short healing time. TRIAL REGISTRATION: Chinese Clinical Trial Register, ChiCTR2000039174. Registered on 28 October 2020.

Promotional effects of Nb5+ and Fe3+ co-doping on catalytic performance and SO2 resistance of MnOx-CeO2 low-temperature denitration catalyst.

As we know, SO2 can cause MnOx-CeO2 (MnCeOx) catalyst poisoning, which seriously shortens the service life of the catalyst. Therefore, to enhance the catalytic activity and SO2 tolerance of MnCeOx catalyst, we modified it by Nb5+ and Fe3+ co-doping. And the physical and chemical properties were characterized. These results illustrate that the Nb5+ and Fe3+ co-doping can optimally improve the denitration activity and N2 selectivity of MnCeOx catalyst at low temperature by improving its surface acidity, surface adsorbed oxygen as well as electronic interaction. What's more, NbOx-FeOx-MnOx-CeO2 (NbFeMnCeOx) catalyst possesses an excellent SO2 resistance due to less SO2 being adsorbed and the ammonium bisulfate (ABS) formed on its surface tends to decompose, as well as fewer sulfate species formed on its surface. Finally, the possible mechanism that Nb5+ and Fe3+ co-doping enhances the SO2 poisoning resistance of MnCeOx catalyst is proposed.

Overexpression of the Capebp2 Gene Encoding the PEBP-like Protein Promotes the Cap Redifferentiation in Cyclocybe aegerita.

Phosphatidylethanolamine-binding protein (PEBP) is widely involved in various physiological behaviors, such as the transition from vegetative growth to reproductive growth in plants, tumorigenesis in the human, etc. However, few functional studies have examined pebp genes affecting the development of fungi. In this study, Capebp2 was cloned from Cyclocybe aegerita AC0007 strains based on the genome sequence and gene prediction, and the sequence alignment of CaPEBP2 with other PEBP proteins from other biological sources including plant, animal, fungi, and bacteria indicated that PEBP had low sequence similarity in fungi, whereas all protein sequences had some conserved motifs such as DPDAP and HRY. Expression analysis showed the transcription level of Capebp2 increased approximately 20-fold in fruiting bodies compared with mycelia. To uncover the function of Capebp2 in C. aegetita development, Capebp2 was cloned into a pATH vector driven by the actin promoter for obtaining overexpression transformant lines. Fruiting experiments showed the transformed strains overexpressing Capebp2 exhibited redifferentiation of the cap on their surface, including intact fruiting bodies or partial lamella during fruiting development stage, and the longitudinal section indicated that all regenerated bodies or lamella sprouted from the flesh and shared the epidermis with the mother fruiting bodies. In summary, the sequence characterization of Capebp2, expression level during different development stages, and function on fruiting body development were documented in this study, and these findings provided a reference to study the role of pebp in the development process of basidiomycetes. Importantly, gene mining of pebp, function characterization, and the regulating pathways involved need to be uncovered in further studies.

HMGB1 mediates synaptic loss and cognitive impairment in an animal model of sepsis-associated encephalopathy.

BACKGROUND: Microglial activation-mediated neuroinflammation is one of the essential pathogenic mechanisms of sepsis-associated encephalopathy (SAE). Mounting evidence suggests that high mobility group box-1 protein (HMGB1) plays a pivotal role in neuroinflammation and SAE, yet the mechanism by which HMGB1 induces cognitive impairment in SAE remains unclear. Therefore, this study aimed to investigate the mechanism of HMGB1 underlying cognitive impairment in SAE. METHODS: An SAE model was established by cecal ligation and puncture (CLP); animals in the sham group underwent cecum exposure alone without ligation and perforation. Mice in the inflachromene (ICM) group were continuously injected with ICM intraperitoneally at a daily dose of 10 mg/kg for 9 days starting 1 h before the CLP operation. The open field, novel object recognition, and Y maze tests were performed on days 14-18 after surgery to assess locomotor activity and cognitive function. HMGB1 secretion, the state of microglia, and neuronal activity were measured by immunofluorescence. Golgi staining was performed to detect changes in neuronal morphology and dendritic spine density. In vitro electrophysiology was performed to detect changes in long-term potentiation (LTP) in the CA1 of the hippocampus. In vivo electrophysiology was performed to detect the changes in neural oscillation of the hippocampus. RESULTS: CLP-induced cognitive impairment was accompanied by increased HMGB1 secretion and microglial activation. The phagocytic capacity of microglia was enhanced, resulting in aberrant pruning of excitatory synapses in the hippocampus. The loss of excitatory synapses reduced neuronal activity, impaired LTP, and decreased theta oscillation in the hippocampus. Inhibiting HMGB1 secretion by ICM treatment reversed these changes. CONCLUSIONS: HMGB1 induces microglial activation, aberrant synaptic pruning, and neuron dysfunction in an animal model of SAE, leading to cognitive impairment. These results suggest that HMGB1 might be a target for SAE treatment.

Organization and Unconventional Integration of the Mating-Type Loci in Morchella Species.

True morels (Morchella spp.) are a group of delicious fungi in high demand worldwide, and some species of morels have been successfully cultivated in recent years. To better understand the sexual reproductive mechanisms of these fungi, we characterized the structure of the mating-type loci from ten morel species, and seven of them were obtained using long-range PCR amplification. Among the studied species, eight were heterothallic, two were homothallic, and four types of composition were observed in the MAT loci. In three of the five black morel species, the MAT1-1-1, MAT1-1-10, and MAT1-1-11 genes were in the MAT1-1 idiomorph, and only the MAT1-2-1 gene was in the MAT1-2 idiomorph, while an integration event occurred in the other two species and resulted in the importation of the MAT1-1-11 gene into the MAT1-2 idiomorph and survival as a truncated fragment in the MAT1-1 idiomorph. However, the MAT1-1-11 gene was not available in the four yellow morels and one blushing morel species. M. rufobrunnea, a representative species of the earliest diverging branch of true morels, along with another yellow morel Mes-15, were confirmed to be homothallic, and the MAT1-1-1, MAT1-1-10, and MAT1-2-1 genes were arranged in a tandem array. Therefore, we hypothesized that homothallism should be the ancestral reproductive state in Morchella. RT-PCR analyses revealed that four mating genes could be constitutively expressed, while the MAT1-1-10 gene underwent alternative splicing to produce different splice variants.

A Putative Guanosine Triphosphate Cyclohydrolase I Named CaGCH1 Is Involved in Hyphal Branching and Fruiting Development in Cyclocybe aegerita.

Guanosine triphosphate (GTP) cyclohydrolase I (GCH1) is the limiting enzyme of the tetrahydrobiopterin (BH4) synthesis pathway. The disruption of gch1 gene may cause conditional lethality due to folic acid auxotrophy in microorganisms, although the function of gch1 in basidiomycetes has not been deciphered so far. In the present study, gch1 expression in Cyclocybe aegerita (cagch1) was downregulated using the RNAi method, which resulted in growth retardation in both solid and liquid medium, with the hyphal tips exhibiting increased branching compared to that in the wild strain. The development of fruiting bodies in the mutant strains was significantly blocked, and there were short and bottle-shaped stipes. The transcriptional profile revealed that the genes of the MAPK pathway may be involved in the regulation of these effects caused by cagch1 knockdown, which provided an opportunity to study the role of gch1 in the development process of basidiomycetes.

Characterization of Vegetative Incompatibility in Morchella importuna and Location of the Related-Genes by Bulk Segregant Analysis.

Vegetative incompatibility (VI) is a widespread phenomenon developed in Morchella importuna, a species of ascomycete fungus that is cultivated on a rapidly expanding scale in China. Understanding the genetic bases of this nonself-recognition phenomenon is beneficial for resolving some problems that are associated with the production of this highly prized edible fungus, such as crossbreeding, strain classification, and pathogen transmission. VI is genetically controlled by het genes, organized in two different systems, namely allelic and nonallelic. These het genes have been well characterized in Podospora anserina and Neurospora crassa. In this work, putative het-homologs were identified in the genome of M. importuna, but their low allelic polymorphism in different vegetative compatibility groups (VCGs) suggested that VI in this fungus might not be regulated by these het genes. The progeny derived from vegetative compatible parents became a VCG, while the single-ascospore strains from vegetative incompatible parents were divided into four VCGs, and the interaction between the inter-group strains led to the formation of two types of barrages, viz., thin dark line and raised aggregate of hyphae. The Bulk Segregant Analysis confirmed that the genes mimpvic32 and mimpvic33 were linked to VI reactions in M. importuna; nevertheless, the formation of barrages also occurred between the pairs carrying the same allele of these two genes. In sum, the VI control system in M. importuna was complicated, and there were more other allelic or non-allelic VI-related genes.

Assessment of the targeted effect of Sijunzi decoction on the colorectal cancer microenvironment via the ESTIMATE algorithm.

OBJECTIVE: Sijunzi decoction (SJZD) was used to treat patients with colorectal cancer (CRC) as an adjuvant method. The aim of the study was to investigate the therapeutic targets and pathways of SJZD towards the tumor microenvironment of CRC via network pharmacology and the ESTIMATE algorithm. METHODS: The ESTIMATE algorithm was used to calculate immune and stromal scores to predict the level of infiltrating immune and stromal cells. The active targets of SJZD were searched in the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and UniProt database. The core targets were obtained by matching the differentially expressed genes in CRC tissues and the targets of SJZD. Then, GO, KEGG and validation in TCGA were carried out. RESULTS: According to the ESTIMATE algorithm and survival analysis, the median survival time of the low stromal score group was significantly higher than that of the high stromal score group (P = 0.018), while the patients showed no significant difference of OS between different immune groups (P = 0.19). A total of 929 genes were upregulated and 115 genes were downregulated between the stromal score groups (|logFC| > 2, adjusted P < 0.05); 357 genes were upregulated and 472 genes were downregulated between the immune score groups. The component-target network included 139 active components and 52 related targets. The core targets were HSPB1, SPP1, IGFBP3, and TGFB1, which were significantly associated with poor prognosis in TCGA validation. GO terms included the response to hypoxia, the extracellular space, protein binding and the TNF signaling pathway. Immunoreaction was the main enriched pathway identified by KEGG analysis. CONCLUSION: The core genes (HSPB1, SPP1, IGFBP3 and TGFB1) affected CRC development and prognosis by regulating hypoxia, protein binding and epithelial-mesenchymal transition in the extracellular matrix.

Purification and Characterization of a Thermo- and pH-Stable Laccase From the Litter-Decomposing Fungus Gymnopus luxurians and Laccase Mediator Systems for Dye Decolorization.

An extracellular laccase (GLL) was purified from fermentation broth of the litter-decomposing fungus Gymnopus luxurians by four chromatography steps, which resulted in a high specific activity of 118.82 U/mg, purification fold of 41.22, and recovery rate of 42.05%. It is a monomeric protein with a molecular weight of 64 kDa and N-terminal amino acid sequence of AIGPV TDLHI, suggesting that GLL is a typical fungal laccase. GLL demonstrated an optimum temperature range of 55°C-65°C and an optimum pH 2.2 toward 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). It displayed considerably high thermostability and pH stability with about 63% activity retained after 24 h at 50°C, and 86% activity retained after 24 h at pH 2.2, respectively. GLL was significantly enhanced in the presence of K+, Na+, and Mg2+ ions. It demonstrated K m of 539 µM and k cat /K m of 140 mM-1⋅s-1 toward ABTS at pH 2.2 and 37°C. Acetosyringone (AS) and syringaldehyde (SA) were the optimal mediators of GLL (0.4 U/ml) for dye decolorization with decolorization rates of about 60%-90% toward 11 of the 14 synthetic dyes. The optimum reaction conditions were determined to be mediator concentration of 0.1 mM, temperature range of 25°C -60°C, and pH 4.0. The purified laccase was the first laccase isolated from genus Gymnopus with high thermostability, pH stability, and effective decolorization toward dyes, suggesting that it has potentials for textile and environmental applications.

Two Cu(II) coordination polymers: Heterogeneous catalytic Knoevenagel condensation reaction and treatment activity on atherosclerosis via regulating the expression of the COX-2 in vascular endothelial cells.

By altering auxiliary nitrogen-donor ligands, two novel coordination polymers (CPs) containing Cu(II) formulated as [Cu2.5(L)(trz)2(H2O)2]·2H2O (1) (Htrz = 1,2,4-triazole and H3L = 5-(4-carboxybenzyloxy)isophthalic acid) and [Cu(HL)(Hbiz)] (2, Hbiz = benzimidazole) have been produced under the hydrothermal conditions. The complex 2 with both acidic and basic sites was investigated as heterogeneous catalyst, which reveals highly efficient catalytic property of the Knoevenagel condensation reaction. Dynamic changes of coagulation parameters during atherosclerosis was also explored via detecting activated partial thromboplastin time (APTT) and prothrombin time (PT), and the results showed that compared with CP 2, CP 1 has a stronger improvement on the coagulation parameters during atherosclerosis. Then, the high-sensitivity C-reactive protein and matrix metalloproteinase-1 released by the atherosclerotic segment was detected with enzyme linked immunosorbent assay (ELISA) detection, which also revealed that CP 1 could obviously decrease the inflammatory mediator released by the atherosclerotic segment, but not CP 2. And, the cyclooxygenase-2 (COX-2) relative expression level in vascular endothelial cells was detected via the real time RT-PCR. The results of the real time reverse transcription-polymerase chain reaction (RT-PCR) indicated that CP 1 has stronger activity on inhibiting the Notch signaling pathway than CP 2. Finally, we got this information, CP 1 has excellent application values on the coagulation parameters during atherosclerosis via regulating the expression of the COX-2 in vascular endothelial cells.

The Genus Acervus from Southwestern China and Northern Thailand.

Acervus (Pyronemataceae, Pezizales) is a saprobic genus in Pezizomycetes, characterized by colored apothecia, subcylindrical to cylindrical asci and guttulate ascospores. We collected four Acervus samples from China and Thailand. Descriptions and illustrations are introduced for all fresh samples. One new record of A. globulosus from Thailand, one new species, A. rufus, two known species, A. epispartius and A. stipitatus from China are reported. Phylogenetic analysis based on five genes, the large subunit rRNA (LSU), the translation elongation factor-1 alpha (tef1-α), the second largest subunit of RNA polymerase II (rpb2), the largest subunit of RNA polymerase II (rpb1), and the small subunit rRNA (SSU), revealed the distinct position of the new species. The new species is set apart by its red apothecia. A key to Acervus species is also given.

Structural variation and phylogenetic analysis of the mating-type locus in the genus Morchella.

True morels (Morchella spp.) are a group of edible fungi that are in high demand worldwide. However, this group of fungi remains poorly understood in terms of its genetic structure, life cycle, and mating system. In the present study, we cloned the MAT locus in Morchella sp. Mes-20 using long-range polymerase chain reaction (PCR) amplification. Our results showed that the MAT1-2 idiomorph in the single-ascospore isolate YAASMCB-3 was 7.5 kb in length, harboring a single MAT1-2-1 gene, whereas the MAT1-1 idiomorph in the isolate YAASMCB-15 was 7.8 kb, carrying both MAT1-1-1 and MAT1-1-10 genes. Mating-type diagnostic assays of single-ascospore populations confirmed that Mes-20 is heterothallic. In addition, 42 collections belonging to 17 morel species (7 black morel species [Elata clade], 9 yellow morel species [Esculenta clade], and 1 species from the Rufobrunnea clade) were used to clone and characterize the MAT1-1-1 and MAT1-2-1 genes for development of the mating gene genealogies. In concordance with the multilocus phylogenetic trees, collections for the respective species were readily distinguished by well-supported lineages in mating gene genealogies. The topologies were consistent within the Elata clade, whereas the branching order and sister-group relationships slightly varied within the Esculenta clade. Our results show that species in the genus Morchella could be delimited by mating gene genealogies.

Characterization of an acidic α-galactosidase from hemp (Cannabis sativa L.) seeds and its application in removal of raffinose family oligosaccharides (RFOs).

An acidic α-galactosidase designated as hemp seed α-galactosidase (HSG) was purified from hemp (Cannabis sativa L.) seeds. By means of chromatographic procedures which involved chromatography on the cation-exchangers CM-cellulose and SP-Sepharose, chromatography on the anion-exchangers DEAE-cellulose and Q-Sepharose, and gel filtration on Superdex 75 using fast protein liquid chromatography, HSG was purified to electrophoretic homogeneity. Results of SDS-PAGE and gel filtration on FPLC Superdex 75 revealed that the enzyme was a monomeric protein with a molecular weight of 38 kDa. Sequences of the inner peptides of the α-galactosidase obtained by MALDI-TOF-MS showed that HSG was a novel α-galactosidase since there was a little similarity to the majority of α-galactosidases recorded in the literature. A pH of 3.0 and a temperature of 50°C were optimal for the activity of the enzyme. The activity of HSG was inhibited by the chemical modification with N-bromosuccinimide (NBS) reagent. HSG contained 16 tryptophan residues and two tryptophan residues on the surface, which were crucial to the α-galactosidase activity. The heavy metal ions Cd2+, Cu2+, Hg2+ and Zn2+ inhibited its activity. The Km and Vmax for the hydrolysis of pNPGal (4-nitrophenyl α-D-galactopyranoside) were respectively 0.008 mM and 68 µM min-1 mg-1. HSG also catalyzed the hydrolysis of raffinose and other natural substrates. Hence the α-galactosidase possesses a tremendous potential for food and feed industries in the elimination of indigestible oligosaccharides from leguminous products.

Purification and characterization of a novel metalloprotease from fruiting bodies of Oudemansiella radicata.

In this study, a 39-kDa metalloprotease was purified from a rare edible mushroom with health-promoting activities, Oudemansiella radicata, using a purification protocol which entailed anion exchange chromatography on DEAE-cellulose and Q-Sepharose columns and gel filtration by fast protein liquid chromatography on a Superdex 75 column. Some peptide sequences were obtained by LC-MS/MS analysis and one of the sequences, DAWIQADVNR, manifested 90% identity to Coprinopsis cinerea metalloprotease. The optimal reaction pH and temperature for Oudemansiella radicata protease were pH 7.0 and 50°C, respectively. The protease was purified 79-fold and demonstrated a specific protease activity of 2.42 U/mg. The Km of the purified protease for the casein substrate was 0.65 mg/mL at pH 7.0 and 50°C. The activity of the protease was inhibited by Cd2+, Hg2+, Cu2+, Pb2+ and Fe3+ ions, but was enhanced by K+, Mn2+ and Fe2+ ions. The marked suppression of the protease activity by EDTA indicates that the protease is a metalloprotease.

Characterization of Non-coding Regions in B Mating Loci of Agrocybe salicacola Groups: Target Sites for B Mating Type Identification.

Agrocybe salicacola is a delicious and cultivable mushroom. It is important to understand this species' inherent characteristics, especially to elucidate the constitution and segregation of mating genes. In this study, two compatible B mating loci in strain YAASM0711 of A. salicacola were cloned from the monokaryons, and sequence and phylogeny analyses showed two conserved genes encoding pheromone receptors maybe lost mating activity, which determined by comparing with those of other mushrooms. In the conserved regions of mating loci, partial insertion/deletion fragments made non-coding regions posses polymorphisms, and monokaryotic strains of different mating types were distinguished from each other according to the amplification profile of variable regions, which suggested mating loci were integrally assigned to offspring strains during mitosis in A. salicacola. As our known, it is the first to develop molecular markers for B mating-type identification using variable non-coding fragments of mating loci in basidiomycetes.

Biochemical characteristics of a novel protease from the basidiomycete Amanita virgineoides.

The characterization of a novel protease from Amanita virgineoides is described. The A. virgineoides protease was purified to homogeneity using Q-Sepharose, carboxymethyl-cellulose, diethylaminoethyl-cellulose, and a gel filtration step on Superdex 75. The molecular mass of the purified protease was estimated to be 16.6 kDa. The protease was purified 32.1-fold, and its specific activity was 301.4 U/mg. The optimum pH was 4.0, and the optimum temperature was 50 °C. Kinetic constants (Km , Vmax ) were determined under the optimum reaction conditions, with Km and Vmax , being 3.74 mg/mL and 9.98 µg mL-1 Min-1 , respectively. The activity of the protease was curtailed by Cu2+ , Pb2+ , Fe3+ , Cd2+ , and Hg2+ ions but enhanced by Mg2+ , Ca2+ , and K+ ions at low concentrations. The protease activity was adversely affected by ethylene diamine tetraacetic acid, suggesting that it is a metalloprotease. Four peptide sequences were obtained from liquid chromatography-tandem mass spectrometry, including KQALSGIR, TIAMDGTEGLVR, VALTGLTVAEYFR, and AGAGSATLSMAYAGAR, which showed 86%, 64%, 60%, and 75% identity with peptides of Hypsizygus marmoreus, Dacryopinax sp. DJM-731 SS1, Trametes versicolor FP-101664 SS1, and Paxillus involutus ATCC 200175, respectively. The newly isolated protease showed good hydrolytic activity and biochemical characteristics, which expanded the knowledge of biologically active proteins and provided further insight on this poisonous fungus.

Degradation of dyes using crude extract and a thermostable and pH-stable laccase isolated from Pleurotus nebrodensis.

Three laccase isoenzymes (Lac1, Lac2 and Lac3) have been purified to homogeneity from Pleurotus nebrodensis in our previous study. Lac2 was shown to be the dominant isoform, capable of oxidizing the majority of laccase substrates and manifesting good thermostability and pH stability. Hence, Lac2 was selected to decolourize structurally different dyes and the colour removal efficiencies of Lac2 and the crude extract of P. nebrodensis were compared. By monitoring the λmax of the reaction system during the course of biotransformation, clear hypsochromic shifts were observed for most of the dyes examined, illustrating that at least one peak disappeared as a result of laccase treatment. In general, Lac2 was more efficient within a short time (1 h) and the crude extract, in general, could achieve similar or even higher efficiency when the duration of treatment was extended to 24 h. Malachite green (MG) was chosen to study the detoxifying potential of Lac2, because of the relatively simple structure and high toxicity of the dye towards microorganisms. The toxicity of MG towards both bacteria (Bacillus subtilis, Bacillus licheniformis, Pseudomonas fluorescens and Escherichia coli) and fungi (Fusarium graminearum and Trichoderma harzianum) was dramatically decreased and the potential mechanism was estimated by GC-MS as to remove four methyl groups firstly and the two newly formed amine groups would be degraded or polymerized further. The present study facilitates an understanding of the application of P. nebrodensis laccases and furnishes evidence for the safety of their utilization in the treatment of wastewater emanating from textile industries.

Species clarification of the culinary Bachu mushroom in western China.

The Bachu mushroom, previously identified as Helvella leucopus, is characterized by a saddle-shaped, to irregularly lobed pileus, with a gray, brown to blackish hymenium and a whitish to pale receptacle surface and white, terete stipe with enlarged basal grooves. It has high economic value, mostly as a dietary supplement in western China, and its medicinal functions have raised broad interest. In the present paper species of the Bachu mushroom in Xinjiang Autonomous Region, western China were investigated with morphology and DNA sequence data. Phylogenetic analyses inferred from ITS, 28S and TEF1 sequence data strongly supported lineages corresponding to morphological features. The Bachu mushroom, which differs from the European Helvella leucopus, comprises two distinct new species, namely Helvella bachu and Helvella subspadicea. In this paper we introduce the new species with descriptions and figures and compare them with similar taxa. The European Helvella spadicea is also re-examined, described and illustrated.