2-Furoic acid associated with the infection of nematodes by Dactylellina haptotyla and its biocontrol potential on plant root-knot nematodes.

Dactylellina haptotyla is a typical nematode-trapping fungus that has garnered the attention of many scholars for its highly effective lethal potential for nematodes. Secondary metabolites play an important role in D. haptotyla-nematode interactions, but which metabolites perform which function remains unclear. We report the metabolic functions based on high-quality, chromosome-level genome assembly of wild D. haptotyla YMF1.03409. The results indicate that a large variety of secondary metabolites and their biosynthetic genes were significantly upregulated during the nematode-trapping stage. In parallel, we identified that 2-furoic acid was specifically produced during nematode trapping by D. haptotyla YMF1.03409 and isolated it from fermentation production. 2-Furoic acid demonstrated strong nematicidal activity with an LD50 value of 55.05 µg/mL against Meloidogyne incognita at 48 h. Furthermore, the pot experiment showed that the number of galls of tomato root was significantly reduced in the experimental group treated with 2-furoic acid. The considerable increase in the 2-furoic acid content during the infection process and its virulent nematicidal activity revealed an essential synergistic effect during the process of nematode-trapping fungal infection. IMPORTANCE Dactylellina haptotyla have significant application potential in nematode biocontrol. In this study, we determined the chromosome-level genome sequence of D. haptotyla YMF1.03409 by long-read sequencing technology. Comparative genomic analysis identified a series of pathogenesis-related genes and revealed significant gene family contraction events during the evolution of D. haptotyla YMF1.03409. Combining transcriptomic and metabolomic data as well as in vitro activity test results, a compound with important application potential in nematode biocontrol, 2-furoic acid, was identified. Our result expanded the genetic resource of D. haptotyla and identified a previously unreported nematicidal small molecule, which provides new options for the development of plant biocontrol agents.

Fecal calprotectin as an indicator in risk stratification of pouchitis following ileal pouch-anal anastomosis for ulcerative colitis.

BACKGROUND: Pouchitis is the most common complication following restorative proctocolectomy and ileal pouch-anal anastomosis (IPAA) for ulcerative colitis (UC). Fecal calprotectin (FC) is a noninvasive indicator of the intestinal inflammatory status. This study was conducted to evaluate the clinical value of the FC concentration for the diagnosis and risk assessment of pouchitis. PATIENTS AND METHODS: This retrospective study involved patients who underwent IPAA for UC at Tianjin Medical University General Hospital from January 2015 to January 2019. The patients were categorized into pouchitis and non-pouchitis groups based on their Pouchitis Disease Activity Index (PDAI) score. Laboratory indicators, including the FC concentration, were collected from both groups. RESULTS: Sixty-six patients with UC after IPAA were included in the study and divided into the non-pouchitis group (n = 40) and pouchitis group (n = 26). The correlation coefficient between the FC concentration and the PDAI score was 0.651 (p < 0.001). Receiver operating characteristic analysis showed that the FC cut-off value for predicting pouchitis was 579.60 µg/g (area under the curve, 0.938). The patients were then divided into three subgroups according to their PDAI score (0-2, 3-6, and ≥7), and significant differences in the FC concentration were found among the three subgroups. The best FC cut-off value for predicting a high risk of pouchitis (PDAI score of 3-6) was 143.25 µg/g (area under the curve, 0.876). CONCLUSIONS: FC is a useful biomarker in patients with pouchitis. Patients are advised to regularly undergo FC measurement to monitor for pouchitis. An FC concentration in the range of 143.25-579.60 µg/g is predictive of a high risk for pouchitis, and further examination and preventive treatment are necessary in such patients.KEY MESSAGESFecal calprotectin can be used to quantify pouch inflammation.Fecal calprotectin can be used to predict a high risk of pouchitis.

Identification and Mechanism of Action of the Global Secondary Metabolism Regulator SaraC in Stereum hirsutum.

DNA methylation is an important factor in the regulation of gene expression. In analyzing genomic data of Stereum hirsutum FP-91666, we found a hypothetical bifunctional transcription regulator/O6Meguanine-DNA methyltransferase (named SaraC), which is widely present in both bacteria and fungi, and confirmed that its function in bacteria is mainly for DNA reparation. In this paper, we confirmed that SaraC has the function of DNA binding and demethylation through surface plasma resonance and reaction experiments in vitro. Then, we achieved the overexpression of SaraC (OES) in S. hirsutum, sequenced the methylation and transcription levels of the whole-genome, and further conducted untargeted metabolomics analyses of the OES transformants and the wild type (WT). The results confirmed that the overall-methylation levels of the transformants were significantly downregulated, and various genes related to secondary metabolism were upregulated. Through comparative untargeted metabolomic analyses, it showed that OES SA6 transformant produced a greater number of hybrid polyketides, and we identified 2 novel hybrid polyketides from the fermentation products of SA6. Our results show that overexpression SaraC can effectively stimulate the expression of secondary-metabolism-related genes, which could be a broad-spectrum tool for discovery of metabolites due to its cross-species conservation. IMPORTANCE Fungi are one of the important sources of active compounds. However, in fungi, most of the secondary metabolic biosynthetic gene clusters are weakly expressed or silenced under conventional culture conditions. How to efficiently excavate potential new compounds contained in fungi is becoming a research hot spot in the world. In this study, we found a DNA demethylation protein (SaraC) and confirmed that it is a global secondary metabolism regulator in Stereum hirsutum FP-91666. In the past, SaraC-like proteins were mainly regarded as DNA repair proteins, but our findings proved that it will be a powerful tool for mining secondary metabolites for overexpression of SaraC, which can effectively stimulate the expression of genes related to secondary metabolism.

Case Report: An Unusual Case of Pulmonary Metastatic Adenocarcinoma From Low-Grade Appendiceal Mucinous Neoplasms.

Background: Low-grade appendiceal mucinous neoplasms (LAMNs) are indolent tumors with low-grade cytology. Although peritoneal dissemination is common due to tumor rupture and mucinous deposits on the visceral peritoneal surface, distant involvement, such as lung, is rarely seen due to lack of invasiveness. Case Presentation: A 70-year-old woman presented to the hospital due to continuously elevated carcinoembryonic antigen (CEA) levels for 10 months without any symptoms. PET/CT revealed two lesions located in the left lung and appendix. The postoperative pathology results revealed pulmonary mucinous adenocarcinoma and LAMN. Then we performed next-generation sequencing (NGS) to clarify the relationship between the two tumors. The sequencing result showed that both tumors harbored the common tumor mutations, KRAS (p.G12D), GNAS (p.R201H), and BRAF (p.R735Q), which indicated that the pulmonary tumor was a metastasis of LAMN. Conclusion: This case is unusual in that the primary LAMN and the pulmonary metastasis are present at the time of diagnosis. This study reported the first pulmonary metastasis from LAMN verified by NGS.

Identification of Nematicidal Metabolites from Purpureocillium lavendulum.

Purpureocillium lavendulum is a fungus with promising biocontrol applications. Here, transcriptome data acquired during the infection of Caenorhabditis elegans by Purpureocillium lavendulum showed that the transcription of metabolite synthesis genes was significantly up-regulated after 24 and 48 h of the fungus-nematode interaction. Then, the up-regulated transcription level of lipoxygenase was confirmed by RT-qPCR. The ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) analysis of differential metabolites revealed that this interaction resulted in the emergence of new metabolites or enhanced the production of metabolites. The results of the UPLC-MS analysis and the nematicidal assay were used to establish optimal culturing conditions under which 12 metabolites, including 3 hydroxylated C18 fatty acids and 9 steroids, were isolated and identified. Among them, hydroxylated fatty acids showed pronounced nematicidal activity against Meloidogyne incognita, and two degradative sterols showed chemotaxis activity to M. incognita. This study lays a foundation for the function of lipoxygenase and its products during the infection of Purpureocillium lavendulum.

Pathogenicity and Metabolites of Purpureocillium lavendulum YMF1.00683 against Meloidogyne incognita.

Purpureocillium lavendulum is a biological control agent with several registered products that can parasitize the eggs and larvae of various pathogenic nematodes. In this study, the pathogenicity and secondary metabolites of the fungus P. lavendulum YMF1.00683 were investigated. The strain YMF1.00683 had infection efficiency against the plant root-knot nematode Meloidogyne incognita. The strain's process of infecting nematodes was observed under a microscope. Moreover, seven metabolites, including a new sterol (1), were isolated and identified from cultures of YMF1.0068 in Sabouraud's dextrose agar. A bioassay showed that 5-methoxymethyl-1H-pyrrole-2-carboxaldehyde (7) is toxic to M. incognita and affects the egg hatching. It caused 98.23% mortality in M. incognita and could inhibit 80.78% of the hatching eggs at 400 µg/mL over a period of 96 h. Furthermore, 5-methoxymethyl-1H-pyrrole-2-carboxaldehyde (7) showed a strong avoidance effect at 40 ppm, and its chemotactic index value was -0.37. The results indicate that P. lavendulum could produce active metabolites against M. incognita.

Solitary Cecal Ulcer: A Case Report.

Background: Solitary cecal ulcer is a rare disease. Its etiology is unknown and there are no pathognomonic symptoms. There are rare reports mimicking carcinoma as seen in this case. Case Presentation: A 64 year-old woman presented with a history of intermittent right lower abdominal pain for 20 years and worsening for 1 year. Colonoscopy revealed an enormous cecal ulcer. The PET-CT showed increased metabolism of the lesion. She underwent a right hemicolectomy. Histopathological examination revealed chronic non-specific inflammation. A rare diagnosis of the solitary cecal ulcer was ultimately made. Conclusion: Solitary cecal ulcer is a rare, idiopathic entity. It mimics inflammatory bowel disease, malignancy, infection, etc. The comprehensive images of this case describe the characteristics of the disease.

Recent Advances in the Synthetic Biology of Natural Drugs.

Natural drugs have been transformed and optimized during the long process of evolution. These compounds play a very important role in the protection of human health and treatment of human diseases. Sustainable approaches to the generation of raw materials for pharmaceutical products have been extensively investigated in drug research and development because chemical synthesis is costly and generates pollution. The present review provides an overview of the recent advances in the synthetic biology of natural drugs. Particular attention is paid to the investigations of drugs that may be mass-produced by the pharmaceutical industry after optimization of the corresponding synthetic systems. The present review describes the reconstruction and optimization of biosynthetic pathways for nine drugs, including seven drugs from plant sources and two drugs from microbial sources, suggesting a new strategy for the large-scale preparation of some rare natural plant metabolites and highly bioactive microbial compounds. Some of the suggested synthetic methods remain in a preliminary exploration stage; however, a number of these methods demonstrated considerable application potential. The authors also discuss the advantages and disadvantages of the application of synthetic biology and various expression systems for heterologous expression of natural drugs. Thus, the present review provides a useful perspective for researchers attempting to use synthetic biology to produce natural drugs.

Visceral-to-Subcutaneous Fat Ratio Is a Potential Predictor of Postoperative Complications in Colorectal Cancer.

BACKGROUND Colorectal cancer (CRC) is one of the most common malignant tumors. Surgery is the primary treatment for CRC. Recent studies have shown that visceral-to-subcutaneous fat ratio (VSR) may be a potential indicator of risk. The aim of the present study was to determine whether VSR is a suitable predictor of the impact of postoperative complications in CRC. MATERIAL AND METHODS Clinical data from 129 patients were analyzed retrospectively. All patients underwent laparoscopic surgery for CRC. Preoperative imaging was used to quantify VSR. The primary outcome was 30-day postoperative complications. Multivariate logistic regression was used to determine the association between obesity indexes and postoperative complications. RESULTS There were 129 patients who underwent primary resections of colorectal cancer. At 30 days after surgery, postoperative complications had occurred in 33 (25.6%) patients. VSR was significantly associated with postoperative complications in multivariate analysis (P=0.032, OR 6.103, 95% CI 1.173-31.748). In ROC analysis, VSR was a potential predictor of complications (AUC 0.650). A cutoff value of VSR ≥0.707 was associated with 60% sensitivity and 29% specificity for postoperative complications. Patients with VSR ≥0.707 had 41.7% risk of morbidity, whereas those with <0.707 had 16.0% risk (P=0.001). CONCLUSIONS Our study shows that VSR is superior to VFA for prediction of the risk of complications following CRC laparoscopic surgery. We have also identified the optimal cutoff values for the use of VSR for this purpose. Measurement of VSR and identifying patients with increased risk of postoperative complications facilitate making perioperative decisions.

Intestinal obstruction in pregnancy with reverse rotation of the midgut: A case report.

BACKGROUND: Reverse rotation of the midgut is a rare type of intestinal malrotation. Volvulus of the right colon or entire midgut, stenosis of the transverse colon and obstruction of the duodenojejunal junction are common complications of reverse rotation. In this study, we report the first case of intestinal obstruction associated with reverse rotation in pregnancy. CASE SUMMARY: A 31-year-old woman at 36+2 wk gestation presented to the emergency department with progressive abdominal cramping, nausea and bilious vomiting. Abdominal ultrasound scanning showed dilatation of the bowel. Computed tomography scanning revealed features of reverse rotation of the midgut with intestinal volvulus. After consultation with the obstetrician, the pregnancy was terminated and exploratory abdominal surgery was performed. Intra-operatively, it was found that the mesentery of the colon and small intestine was insufficiently attached. The right colon and the small intestinal mesentery was twisted, and intestinal necrosis was observed. The duodenum and duodenojejunal junction were curved in front of the transverse colon, and the transverse colon passed through the tunnel behind the mesenteric root. Intestinal reverse rotation with volvulus was confirmed. The |necrotic intestine was resected and small intestine mesenteric reconstruction was performed. The patient recovered after surgery. After leaving the hospital, the patient and her daughter remained well during an 8-month follow-up period. CONCLUSION: We report the diagnosis, treatment and etiology of a pregnant patient with intestinal obstruction due to reverse rotation of the midgut. For similar cases, appropriate diagnosis and treatment should be carried out according to the condition of the fetus and pregnant woman.

Gastroduodenitis associated with ulcerative colitis: A case report.

BACKGROUND: Ulcerative colitis (UC) is defined as a chronic inflammatory bowel disease that can occur in any part of the large bowel. In addition, UC affects only the large bowel except for backwash ileitis and pouchitis, whereas Crohn's disease (CD) affects the entire digestive tract. Inflammatory bowel disease (IBD) patients tend to be diagnosed with CD or indeterminate colitis when combined with gastric lesion. However, in recent years, some UC patients are reported to have various degrees of lesions in gastroduodenum. Here, we report a case of gastroduodenitis associated with UC (GDUC). CASE SUMMARY: A 25-year-old man with a history of Klippel-Trenaunay syndrome presented to the hospital with mucopurulent bloody stool and epigastric persistent colic pain for 2 wk. Continuous superficial ulcers and spontaneous bleeding were observed under colonoscopy. Subsequent gastroscopy revealed mucosa with diffuse edema, ulcers, errhysis, and granular and friable changes in the stomach and duodenal bulb, which were similar to the appearance of the rectum. After ruling out other possibilities according to a series of examinations, a diagnosis of GDUC was considered. The patient hesitated about intravenous corticosteroids, so he received a standardized treatment with pentasa of 3.2 g/d. After 0.5 mo of treatment, the patient's symptoms achieved complete remission. Follow-up endoscopy and imaging findings showed no evidence of recurrence for 26 mo. CONCLUSION: The occurrence of gastrointestinal involvement in UC is rare, which may open a new window for studying the etiology and pathogenesis of UC. Physicians should consider broad differential diagnosis by endoscopic biopsy and laboratory examinations.