Ena Proteins Respond to PacC-Mediated pH Signaling Pathway and Play a Crucial Role in Patulin Biosynthesis.

Penicillium expansum is a main producer of patulin that causes severe postharvest decay and food safety issues in the fruit industry. Development, pathogenicity, and patulin production of P. expansum are strongly influenced by the PacC-pH signaling pathway. Global transcription factor PacC regulates various fungal biological processes through a complicated molecular network. In the present study, three Ena family genes (PeEnas), PeEnaA, PeEnaB, and PeEnaC, as important downstream targets of PePacC, were identified in P. expansum. Deletion of PeEnaA, PeEnaB, and PeEnaC showed little effect on mycelial growth under alkaline or high salinity conditions, but double and triple deletion of these genes impaired the virulence of P. expansum on apple fruit. Notably, patulin biosynthesis of P. expansum was distinctly inhibited in the deletion mutants of PeEnas. PeEnas regulated expressions of the patulin gene cluster, AP1, CreA, Sge1, and Hog1 at the transcriptional level and played roles in maintaining membrane potential. Overexpression of PeEnaC in ΔPePacC restored the patulin production defect of ΔPePacC. Our results indicated that, as downstream targets of PePacC, the PeEna family proteins play a crucial role in patulin biosynthesis in P. expansum.

Immobilized short-chain dehydrogenase/reductase on Fe3O4 particles acts as a magnetically recoverable biocatalyst component in patulin bio-detoxification system.

Patulin is one of the most important mycotoxins that contaminates fruit-derived products and causes acute or chronic toxicity in humans. In the present study, a novel patulin-degrading enzyme preparation was developed by taking a short-chain dehydrogenase/reductase and covalently linking it to dopamine/polyethyleneimine co-deposited magnetic Fe3O4 particles. Optimum immobilization provided 63% immobilization efficiency and 62% activity recovery. Moreover, the immobilization protocol substantially improved thermal and storage stabilities, proteolysis resistance, and reusability. Using reduced nicotinamide adenine dinucleotide phosphate as a cofactor, the immobilized enzyme exhibited a detoxification rate of 100% in phosphate-buffered saline and a detoxification rate of more than 80% in apple juice. The immobilized enzyme did not cause adverse effects on juice quality and could be magnetically separated quickly after detoxification to ensure convenient recycling. Moreover, it did not exhibit cytotoxicity against a human gastric mucosal epithelial cell line at a concentration of 100 mg/L. Consequently, the immobilized enzyme as a biocatalyst had the characteristics of high efficiency, stability, safety, and easy separation, establishing the first step in building a bio-detoxification system to control patulin contamination in juice and beverage products.

Interleukin 37 inhibits the migration and invasion of Glioma cells.

Glioma is a medical term that describes a tumor originating in the brain. Several risk factors could develop glioma such as occupational exposure, gene mutation and ionizing radiation. Therefore, we aim to determine the expression and biological function of interleukin 37 (IL-37) in gliomas with different pathological grades. We used 95 participants with different pathological grades of glioma as our data subjects. We used CCK-8 assay and transwell assay to explore the proliferation of U251 over-expressing IL-37 and migration and invasion of U251. We found that IL-37 expression in tumor tissues was significantly higher than in normal tissue. The reduced IL-37 expression in gliomas was significantly associated with a higher WHO grade and lower Karnofsky Performance Status score. IL-37 expression in glioma tissues showed a decline with the increase of the WHO glioma grade. Patients with low IL-37 expression showed a shorter median survival. Transwell assay indicated that migration and invasion of U251 over-expressing IL-37 was significantly lower than that of the control at 24 h. Our findings showed that low IL-37 expression was negatively correlated with pathological grade and was positively correlated with survival time.

Effects and mechanisms of plant bioactive compounds in preventing fungal spoilage and mycotoxin contamination in postharvest fruits: A review.

Spoilage and mycotoxin contamination of fruits cause significant economic losses and food safety issues. Synthetic chemical fungicide treatment as primary postharvest management has attracted increasing public concern in recent years, because it may cause negative effects on the environment and human health. Numerous bioactive compounds from plants have demonstrated excellent control effects on fruit spoilage and mycotoxin contamination. Plant bioactive compounds have been considered one of the most promising alternatives, because they are generally regarded as safe and environmentally friendly. Here, we reviewed the most recent advances in plant bioactive compounds in the prevention of fungal spoilage and mycotoxin contamination in fruits. The control effects of these compounds and the mechanisms involved were summarized, and current limitations and future perspectives were discussed.

Multi-layer reconstruction of skull base after endoscopic transnasal surgery for invasive pituitary adenomas.

OBJECTIVE: To explore the efficacy of multi-layer skull base reconstruction after endoscopic transnasal surgery for invasive pituitary adenomas (IPAs). CLINICAL RATIONALE FOR THE STUDY: Skull base reconstruction for IPAs. MATERIAL AND METHODS: This retrospective analysis involved 160 patients with IPAs who underwent operations from October 2018 to October 2020. All patients were diagnosed with IPAs by pituitary enhanced magnetic resonance imaging, and all tumours were confirmed to be Knosp grades 3a, 3b, or 4. The experimental group and the control group comprised 80 patients in each, and we used different methods to reconstruct the skull base in each group. The comparison indicators included cerebrospinal fluid leakage, sellar floor bone flap (or middle turbinate) shifting, delayed healing of the skull base reconstructed tissue, nasal discomfort, and epistaxis. We used the chi-square test, and p < 0.05 was considered statistically significant. RESULTS: In the experimental group, cerebrospinal fluid leakage occurred intraoperatively in 73 patients, two of whom had cerebrospinal fluid leakage postoperatively. Brain CT 12 months postoperatively showed no sellar floor bone flap (or middle turbinate) shifting. Endoscopic transnasal checks performed seven days after surgery showed that the skull base reconstructed tissue had healed in 74 patients and had failed to heal in six. However, endoscopic transnasal checks showed that all six of these patients' pedicled nasoseptal flaps had healed well by 14 days after surgery. Other sequelae comprised nasal discomfort in four patients, and epistaxis in four. In the control group, cerebrospinal fluid leakage occurred intraoperatively in 71 patients, 14 of whom had cerebrospinal fluid leakage postoperatively. Brain CT 12 months postoperatively showed floor bone flap (or middle turbinate) shifting in 12 patients. Endoscopic transnasal checks performed seven days after surgery showed that the skull base reconstructed tissue had healed in 65 patients. In 12 patients, pedicled nasoseptal flaps had healed well by 14 days after surgery, while the remaining three patients required reoperation. Other sequelae comprised nasal discomfort in five patients, and epistaxis in six. CONCLUSIONS: This new method of multi-layer skull base reconstruction could play an important role in endoscopic transnasal IPA surgery.

A MADS-box transcription factor, SlMADS1, interacts with SlMACROCALYX to regulate tomato sepal growth.

In flowering plants, sepals play important roles in the development of flowers and fruit, and both processes are regulated by MADS-box (MADS) transcription factors (TFs). SlMADS1 was previously reported to act as a negative regulator of fruit ripening. In this study, expression analysis shown that its transcripts were very highly expressed during the development of sepals. To test the role of SlMADS1, we generated KO-SlMADS1 (knock-out) tomato mutants by CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9) technology and over-expression of SlMADS1 (OE-SlMADS1). The sepals and individual cells of KO-SlMADS1 mutants were significantly elongated, compared with the wild type (WT), whereas the sepals of OE-SlMADS1 tomatoes were significantly shorter and their cells were wider. RNA-seq (RNA-sequencing) of sepal samples showed that ethylene-, gibberellin-, auxin-, cytokinin- and cell wall metabolism-related genes were significantly affected in both KO-SlMADS1 and OE-SlMADS1 plants with altered sepal size. Since SlMACROCALYX (MC) is known to regulate the development of tomato sepals, we also studied the relationship between SlMC and SlMADS1 and the result showed that SlMADS1 interacts directly with SlMC. In addition, we also found that manipulating SlMADS1 expression alters the development of tomato plant leaves, roots and plant height. These results enrich our understanding of sepal development and the function of SlMADS1 throughout the plant.

Highly efficient removal of patulin using immobilized enzymes of Pseudomonas aeruginosa TF-06 entrapped in calcium alginate beads.

Patulin is a toxic secondary metabolite produced by several moulds, which contaminates fruits and their products posing serious threats to human health. Though several microorganisms and enzymes have been reported to effectively degrade patulin, separation of them from fruit juice challenges the commercial applications. Here, a Pseudomonas aeruginosa strain TF-06 was isolated, its patulin degradation mechanism and optimum conditions for enzyme immobilization were investigated. The results indicated that TF-06 could degrade patulin into non-cytotoxic E/Z-ascladiol mainly by the activity of intracellular enzymes. For easy separation of enzymes, calcium alginate was selected for immobilization of intracellular enzymes from TF-06. The immobilized enzyme beads were effective in detoxification of patulin in apple juice. The mitigation rate was reached 95%, while there was no negative effect on juice quality. The study provides a promising way to resolve the issue of enzyme separation during mycotoxin biological detoxification in fruit juice.

Sellar Floor Bone Flap With a Pedicled Nasoseptal Flap in Endoscopic Transnasal Pituitary Adenoma Surgery.

OBJECTIVE: To explore the clinical effect of sellar floor bone flap with a pedicled nasoseptal flap in endoscopic transnasal pituitary adenoma surgery for skull base reconstruction.Method: This was a retrospective clinical analysis of 30 patients with pituitary adenoma operated by the same neurosurgical team from June 2015 to June 2018. All patients were diagnosed with pituitary adenoma by pituitary magnetic resonance imaging, and the authors confirmed that the sellar floor bone was intact using sphenoid sinus computed tomography. All patients underwent an endoscopic transnasal approach, and the authors created a pedicled nasoseptal flap and sellar floor bone flap intraoperatively and reconstructed the skull base at the end of the surgery. Postoperative complications constituted cerebrospinal fluid leakage, brain tissue herniation, nasal discomfort, decreased sense of smell, and epistaxis. RESULTS: Cerebrospinal fluid leakage occurred in 13 patients (43.3%) intraoperatively; small amounts in 6 patients (20.0%), moderate amounts in 3 patients (10.0%), and large amounts in 4 patients (13.3%). Only 1 patient (3.3%) with large-volume cerebrospinal fluid leakage intraoperatively experienced cerebrospinal fluid leakage postoperatively, and this resolved with lumbar catheter drainage and bed rest. The 6-month postoperative follow-up brain computed tomography findings revealed brain tissue herniation in no patients, nasal discomfort in 3 patients (10.0%), decreased sense of smell in 5 patients (16.7%), and epistaxis in 2 patients (6.7%). CONCLUSION: Reconstructing the skull base with a sellar floor bone flap and a pedicled nasoseptal flap played an important role in preventing cerebrospinal fluid leakage and brain tissue herniation in endoscopic transnasal pituitary adenoma surgery and did not increase the incidence of postoperative nasal discomfort, decreased sense of smell, or epistaxis.

Characterization of a short-chain dehydrogenase/reductase and its function in patulin biodegradation in apple juice.

Biodegradation based on microbial enzymes is considered to be one of the promising ways for controlling patulin contamination. However, few patulin degrading enzymes have been isolated and characterized until now. Here, a short-chain dehydrogenase/reductase (SDR) gene, CgSDR, was cloned from a yeast strain Candida guilliermondii, and expressed in Escherichia coli. The expression of CgSDR conferred a strong patulin tolerance and degradation ability to E. coli, and purified CgSDR could transform patulin into E-ascladiol in vitro with NADPH as a coenzyme. Moreover, addition of CgSDR at 150 µg/mL could reduce 80% of patulin in apple juice and the biodegradation process did not affect the quality of the apple juice. A molecular docking analysis and site-directed mutagenesis indicated that CgSDR might interact with patulin via VAL188 as an active binding sites. The findings provide new insights for developing enzymic formulations for mycotoxin detoxification in fruit derived products.

Ribonucleoside Diphosphate Reductase Plays an Important Role in Patulin Degradation by Enterobacter cloacae subsp. dissolvens.

Patulin contamination is a worldwide concern due to its significant impact on human health. Several yeast strains have been screened for patulin biodegradation; however, little information is available on bacterial strains and their mechanism of degradation. In the present study, we isolated a bacterial strain TT-09 and identified it as Enterobacter cloacae subsp. dissolvens based on the BioLog system and 16S rDNA phylogenetic analysis. The strain was demonstrated to be able to transform patulin into E-ascladiol. Isobaric tags for relative and absolute quantitation and reverse transcription quantitative polymerase chain reaction analyses provided evidence that ribonucleoside diphosphate reductase (NrdA), an important enzyme involved in DNA biosynthesis, plays a crucial role in patulin degradation. Deletion of nrdA resulted in a total loss in the ability to degrade patulin in TT-09. These results indicate a new function for NrdA in mycotoxin biodegradation. The present study provides evidence for understanding a new mechanism of patulin degradation and information that can be used to develop new approaches for managing patulin contamination.