Screening, identification, and characterization of molds for brewing rice wine: Scale-up production in a bioreactor.

Rice wine, well known for its unique flavor, rich nutritional value, and health benefits, has potential for extensive market development. Rhizopus and Aspergillus are among several microorganisms used in rice wine brewing and are crucial for determining rice wine quality. The strains were isolated via Rose Bengal and starch as a combined separation medium, followed by oenological property and sensory evaluation screening. The strain exhibiting the best performance can be screened using the traditional rice wine Qu. The strains YM-8, YM-10, and YM-16, which exhibited strong saccharification and fermentation performance along with good flavor and taste, were obtained from traditional rice wine Qu. Based on ITS genetic sequence analysis, the YM-8, YM-10, and YM-16 strains were identified as Rhizopus microsporus, Rhizopus arrhizus, and Aspergillus oryzae. The optimum growth temperature of each of the three strains was 30°C, 32°C, and 30°C, and the optimum initial pH was 6.0, 6.5, and 6.5, respectively. The activities of α-amylase, glucoamylase, and protease of YM-16 were highest at 220.23±1.88, 1,269.04±30.32, and 175.16±1.81 U/g, respectively. The amino acid content of rice wine fermented in a 20-L bioreactor with the three mold strains was higher than that of the control group, except for arginine, which was significantly lower than that of the control group. The total amino acid content and the total content of each type of amino acid were ranked as YM-16 > YM-8 > YM-10 > control group, and the amino acid content varied greatly among the strains. The control group had a higher content, whereas YM-8 and YM-16 had lower contents of volatile aroma components than the control group and had the basic flavor substances needed for rice wine, which is conducive to the formation of rice wine aroma. This selected strain, YM-16, has strong saccharification and fermentation ability, is a rich enzyme system, and improves the flavor of rice wine, thereby demonstrating its suitability as a production strain for brewing.

Effect of modified starches on the quality of skins of glutinous rice dumplings.

This study aimed to investigate the influence of modified starches on the quality of skins of glutinous rice dumplings (SGRDs), including changes in textural properties, pasting parameters, microstructure, color, transparency, and sensory quality. The results showed that the addition of a single acetylated-modified cassava or potato starch or composite modified cassava and potato starch in a ratio of 2:1 can improve the quality of SGRDs. The springiness and lightness of SGRDs increased, and the transparency increased from 3.22 % to 6.18 %. The cooked samples had delicate mouth-feel, uniform color and luster, good transparency, no depression, and low weight loss and did not stick to the teeth. Moreover, the total consumer acceptability score increased from 60.67 to 89.33, indicating that these products were widely accepted by consumers. However, the addition of hydroxypropyl-modified cassava starch or its composite with other two modified starches had no apparent effect on the quality of SGRDs. In conclusion, the quality of SGRDs were significantly improved by the addition of single or composite acetylated-modified starches. This study provides a theoretical basis for improving the quality of SGRDs.

Isolation of Aroma-Producing Wickerhamomyces anomalus Yeast and Analysis of Its Typical Flavoring Metabolites.

In this study, 21 strains of aroma-producing yeast were isolated from Sichuan paocai juice of farmers in western, eastern and southern Sichuan. One strain, Y3, with the best aroma-producing characteristics, was screened using an olfactory method and a total ester titration method, and was identified as Wickerhamomyces anomalus. The total ester content of Y3 fermentation broth was as high as 1.22 g/L, and there was no white colonies or film on the surface. Meanwhile, the Y3 strain could tolerate 14% salt concentration conditions and grow well in a pH range of 3-4. Through sensory analysis, the fermented mustard with a ratio of Lactiplantibacillus plantarum to Y3 of 1:1 showed the highest overall acceptability. Ethyl acetate with its fruit and wine flavor was also detected in the fermented Sichuan paocai juice with a mixed bacteria ratio of 1:1, analyzed with SPME-GC-MS technology, as well as phenylethyl alcohol, isobutyl alcohol, isothiocyanate eaters, myrcene and dimethyl disulfide. These contributed greatly to the unique flavor of Sichuan paocai. In general, Wickerhamomyces anomalus Y3 enhanced the aroma of the fermented Sichuan paocai.

Characterization of indigenous coagulase-negative staphylococci isolated from Chinese spontaneously fermented meat products.

Technological, safety-related and volatile properties were analyzed in coagulase-negative staphylococci (CNS) isolates from Chinese spontaneously fermented meat products. A total of 107 CNS isolates were identified via 16 S rRNA sequencing, and the most recovered species were S. saprophyticus (53.3 %), S. edaphicus (12.1 %), and S. epidermidis (10.3 %). Among them, 58 CNS isolates belonging to 9 species were selected with higher activities of catalase, nitrate reductase, proteolysis, and lipolysis, as well as higher tolerance to stressful environmental conditions. Then, 7 CNS isolates belonging to 4 species were further selected based upon excellent technological characteristics, lack of hemolysis and antibiotic resistance, and a low production of biogenic amines. The volatile profiles of these 7 strains cultivated in pork broth was determined. S. casei No. 1 produced significant amounts of phenethyl alcohol, geraniol, and 3-methyl-butanol. S. xylosus No. 120 produced the highest amount of methyl ketones with the potential to provide dry-cured odor of fermented meats. The volatile profile was highly strain dependent. Several CNS identified in this study have the potential to be used as the starter cultures for fermented meat products.

Associations between ERAP1 polymorphisms and ankylosing spondylitis susceptibility in HLA-B27 positive population: a Meta-analysis and bioinformatics analysis.

Human leukocyte antigen (HLA)-B27 confers a key role in ankylosing spondylitis (AS) susceptibility. Endoplasmic reticulum aminopeptidase 1 (ERAP1) polymorphisms are associated with AS susceptibility in common population. In this study we intended to evaluate the possible association between ERAP1 polymorphisms and AS susceptibility in HLA-27 positive population. Data were collected from Pubmed, Embase, and Cochrane databases. The pooled odds ratios and 95% confidence intervals of the minor allele of each locus were calculated to appraise the associations under ERAP1 polymorphisms and AS in HLA-B27 positive population. Bioinformatics analysis was performed to explore the underlying mechanism. Four studies were included in this meta-analysis. There was a significant association between the minor allele of rs2287987 and reducing the risk of developing AS in HLA-B27 positive population. But there was no significant association between the minor allele of rs30187, rs27044, rs10050860 and rs17482078 and AS susceptibility. According to HaploReg, 5 motifs changed for rs2287987 were found. The eQTL analysis demonstrated that rs2287987 may influence ERAP1 expression. Rs2287987 in ERAP1 may have small influence on AS susceptibility in HLA-B27 positive population. Bioinformatics analysis indicated that the altered motifs and the change of EARP1 expression may influence the AS susceptibility.

Identification of Two Myrosinases from a Leclercia adecarboxylata Strain and Investigation of Its Tolerance Mechanism to Glucosinolate Hydrolysate.

Glucosinolates (GSLs), secondary metabolites synthesized by cruciferous plants, can be hydrolyzed by myrosinase into compounds, such as isothiocyanates (ITCs), with various bioactivities. Thus, myrosinase plays an important role in the utilization of GSLs. We isolated a bacterial strain, which was identified as Leclercia adecarboxylata, from the rhizosphere soil of rape seedlings and identified two myrosinase genes and an ITC hydrolase gene. Both myrosinases are intracellular and have 658 amino acid residues. Via molecular docking and chemical modification assays investigating the active sites of the myrosinases, arginine was found to be essential for their catalytic activity. Transcriptomic analysis of the response to sinigrin revealed significant up-regulation of some genes involved in allyl-ITC detoxification, with metallo-ß-lactamase 3836 having the highest fold change. Thus, we discovered two myrosinases from L. adecarboxylata and demonstrated that the mechanism of tolerance of the bacterium to allyl-ITC likely involved metallo-ß-lactamase activity.

LncRNA NORAD/miR-202-5p regulates the drug resistance of A549/DDP to cisplatin by targeting P-gp.

In this study, we established the DDP-resistant NSCLC cell line A549/DDP to detect the effect of NORAD on cisplatin resistance of A549/DDP cells. NORAD was highly expressed in A549/DDP cells compared with A549 cells. The MTT data showed that knockdown of NORAD enhanced the inhibition rate of cisplatin on the A549/DDP cells and decreased IC50 value. The colony formation and MTT assay suggested that cisplatin inhibited cell proliferation, and knockdown of NORAD enhanced the inhibitory effect of cisplatin on A549/DDP cells. Besides, we found that NORAD silence reduced the P-gp expression but not BCRP, LRP and MRP. Moreover, NORAD could directly bind to miR-202-5p, and ABCB1 was a target of miR-202-5p. The MTT assay found that miR-202-5p inhibitor reversed the effects of NORAD silence on cisplatin resistance of A549/DDP cells. Then, the Western blot data showed that knockdown of NORAD reduced P-gp expression, and miR-202-5p inhibitor enhanced P-gp expression. ABCB1 overexpression reversed the inhibitory effect of NORAD knockdown on A549/DDP cells. Moreover, NORAD could directly bind to miR-202-5p, and ABCB1 was a target of miR-202-5p. Inhibition of miR-202-5p and overexpression of ABCB1 eliminated the effects of NORAD silence on cisplatin resistance of A549/DDP cells. Overexpression of miR-202-5p suppressed P-gp expression in A549/DDP cells. Collectively, our data showed that NORAD could enhance the DDP resistance of A549/DDP cells and potentially increased P-gp expression by sponging the miR-202-5p.

Correction to: identification of multiple cancer-associated myositis-specific autoantibodies in idiopathic inflammatory myopathies: a large longitudinal cohort study.

After publication of the article [1], it has been brought to our attention that the labels in Fig. 2b have been switched and are as a result incorrect. The label for the red line should have the label "non-CAM" and the yellow line "CAM".

Increased Levels of Soluble Programmed Death Ligand 1 Associate with Malignancy in Patients with Dermatomyositis.

OBJECTIVE: To investigate the levels of soluble programmed death ligand 1 (sPD-L1) and evaluate its association with malignancy in patients with dermatomyositis (DM). METHODS: Levels of sPD-L1 were measured in serum from 88 DM patients without malignancies (sDM), 40 with cancer-related DM (CRDM), and 30 healthy controls (HC) using ELISA. The CRDM subjects were divided into new-onset cancers (nCRDM) and stable cancers (sCRDM). Receiver-operating characteristic (ROC) curve analysis was performed to determine the cutoff sPD-L1 value that distinguished patients with nCRDM from those who were sDM. Serum antitranscriptional intermediary factor 1-γ (TIF1-γ) antibodies were detected using immunoblot, and the diagnostic values for malignancy were compared with sPD-L1 levels in patients with DM. RESULTS: Serum sPD-L1 levels were significantly higher in sDM [median 12.3 ng/ml, interquartile range (IQR) 8.4-16.2] than in HC (median 1.3 ng/ml, IQR 0.4-2.2, p = 0.0001). Extremely high sPD-L1 levels were seen in nCRDM (median 18.5 ng/ml, IQR 13.8-22.4), much higher than those in sCRDM (median 8.5 ng/ml, IQR 6.8-11.8, p = 0.0001). The sPD-L1 levels in 4 patients with nCRDM decreased after curative cancer treatment (p = 0.013). ROC curve analysis revealed that the sPD-L1 value distinguishing nCRDM from sDM was 16.1 ng/ml, with an area under the curve value of 0.72 ± 0.04 (p = 0.0001). The combination of sPD-L1 and anti-TIF1-γ antibodies yielded greater specificity and positive predictive value in diagnosing cancer, reaching values of 95% and 70%, respectively. CONCLUSION: Serum sPD-L1 levels increased significantly in sDM, and markedly high sPD-L1 levels could be a diagnostic indicator for malignancies in patients with DM, especially in those with anti-TIF1-γ antibodies.

Identification of multiple cancer-associated myositis-specific autoantibodies in idiopathic inflammatory myopathies: a large longitudinal cohort study.

BACKGROUND: Cancer is a significant complication contributing to increased mortality in idiopathic inflammatory myopathies (IIMs), and the association between IIMs and cancer has been extensively reported. Myositis-specific autoantibodies (MSAs) can help to stratify patients into more homogeneous groups and may be used as a biomarker for cancer-associated myositis. In this study, we aimed to systematically define the cancer-associated MSAs in IIMs. METHODS: Serum from 627 patients with IIMs was tested for MSAs. The cancer risk with different MSAs was estimated by standardized incidence ratio (SIR). Paraneoplastic manifestation, such as the close temporal relationship between myositis onset and cancer diagnoses in patients with different MSAs, was also evaluated. RESULTS: Compared with the general Chinese population, patients with IIMs and anti-transcriptional intermediary factor (TIF1)-γ antibodies (SIR = 17.28, 95% CI 11.94 to 24.14), anti-nuclear matrix protein (NXP2) antibodies (SIR = 8.14, 95% CI 1.63 to 23.86), or anti-SAE1 antibodies (SIR = 12.92, 95% CI 3.23 to 32.94), or who were MSAs-negative (SIR = 3.99, 95% CI 1.96 to 7.14) faced increased risk of cancer. There was no association between specific MSAs subtypes and certain types of cancer. Paraneoplastic manifestations were observed in the patients carrying anti-TIF1-γ, as well as other MSAs. There were no prognostic differences among the patients with cancer-associated myositis (CAM) from different MSAs subgroups. However, in comparison to those with cancer unrelated to myositis, CAM had a worse prognosis, with an age-adjusted and sex-adjusted Cox hazard ratio (HR) of 10.8 (95% CI 1.38-84.5, p = 0.02) for all-cause mortality. CONCLUSIONS: Our study demonstrates in what is, to our knowledge, the largest population examined to date, that anti-SAE1, and previously reported anti-TIF1-γ and anti-NXP2 antibodies, are all associated with an increased risk of cancer in patients with IIMs. Moreover, our data suggest that in some cases, anti-HMGCR, anti-Jo-1 and anti-PL-12 antibody production might also be driven by malignancy. This can aid in the etiologic research of paraneoplastic myositis and clinical management.

The clinical utility of serum IL-35 in patients with polymyositis and dermatomyositis.

The objectives of this study are to assess the levels of serum Interleukin-35 (IL-35) in patients with idiopathic inflammatory myopathies (IIMs) and to evaluate the association between IL-35 levels and IIM-related features. Serum IL-35 was detected in 76 patients with dermatomyositis (DM), 28 patients with polymyositis (PM), 98 disease controls (40 rheumatoid arthritis (RA), 34 systemic lupus erythematosus (SLE), 12 systemic sclerosis (SSc), and 12 sjogren syndrome (SS)), and 43 healthy controls by ELISA. Follow-up was conducted on 34 patients. Serum IL-35 was higher in myositis (PM/DM) patients than in healthy controls (median 76.6 pg/ml [interquartile range (IQR) 57.9-136.2] vs. 29.9 pg/ml (IQR 21.9-65.5), P < 0.001) and disease controls. Serum IL-35 in IIM patients negatively correlated with disease duration moderately (r = -0.35, P < 0.01). Patients with dysphagia had higher IL-35 than those without (median149.35 pg/ml (IQR 87.97-267.32) vs. 70.72 pg/ml (IQR 54.49-123.42), P = 0.001). Cross-sectional correlation analysis showed a weak positive correlation between serum IL-35 and CK (r = 0.293, P = 0.003), moderate positive correlation with erythrocyte sedimentation rate (ESR) (r = 0.304, P = 0.002), serum ferritin (SF) (r = 0.467, P = 0.001) and LDH levels (r = 0.401, P < 0.001). Additionally, serum IL-35 was higher in patients who were positive for anti-HMGCR (median 292.04 pg/ml (IQR 67.9-442.86) vs. 74.66 pg/ml (IQR 57.24-131.32), P = 0.038) and anti-SRP antibody (median 130.33 pg/ml (IQR 88.04-481.28) vs. 73.06 pg/ml (IQR 56.78-134.28), P = 0.009) than in negative patients, respectively. Follow-up study showed that changes in IL-35 levels after treatment correlated with changes in MYOACT scores moderately (r = 0.375, P = 0.029). These data indicate that increased serum IL-35 could act as a disease activity marker and as a risk factor for esophageal involvement in IIM. IL-35 may participate in the pathophysiological processes of IIM, but it still needs further study to confirm.

Diagnostic and prognostic value of serum thioredoxin and DJ-1 in non-small cell lung carcinoma patients.

In response to reactive oxygen species (ROS), thioredoxin and DJ-1 are upregulated to counteract the detrimental effect of ROS under normal condition. However, cancer cells can take advantage of thioredoxin and DJ-1 against ROS-induced cell damage. In several human cancer types, thioredoxin and DJ-1 were found to be overexpressed. The present study aimed to explore the serum levels of thioredoxin and DJ-1 in non-small cell lung cancer (NSCLC) patients and its relationship to the diagnosis and prognosis of this particular malignancy. Sera from 134 NSCLC patients and 168 healthy controls were obtained. Using the enzyme-linked immunosorbent assay (ELISA) method, the levels of serum thioredoxin and DJ-1 were measured and correlated to the clinicopathological characteristics of NSCLC patients. The diagnostic and prognostic significance of the biomarkers were evaluated by using receiver operating curve (ROC), Kaplan-Meier curve, and log-rank analyses and the Cox proportional hazard model, respectively. Serum thioredoxin and DJ-1 levels were significantly higher in the NSCLC patients than that in the controls (23.5 ± 6.57 vs. 13.8 ± 2.49 and 7.11 ± 2.02 vs. 5.18 ± 1.26, respectively). NSCLC patients at later stage cancer showed significantly higher levels of serum thioredoxin and DJ-1 than those at the early stages (P < 0.01 and P < 0.05, respectively). Multivariate logistic regression analysis showed that high serum thioredoxin level was an independent risk factor for lymph nodal metastases and distant metastases (OR = 2.18, 95 % CI 1.26-3.41 and OR = 3.68, 95 % CI 2.16-5.33, respectively). In addition, an increase in the serum DJ-1 level was also identified as an independent risk factor for nodal metastases (OR = 1.37, 95 % CI 1.11-3.04). For predicting the development of NSCLC, ROC/area under the curve (AUC) analysis for thioredoxin indicated an AUC of 0.80 (sensitivity 0.62, specificity 0.92), and ROC/AUC analysis for DJ-1 showed an AUC of 0.78 (sensitivity 0.66, specificity 0.89). NSCLC patients with high serum thioredoxin and DJ-1 levels had lower survival rates than those with low levels, and multivariate analyses for overall survival revealed that high serum thioredoxin levels served as an independent prognostic factor for NSCLC (HR = 2.07, 95 % CI 1.19-3.48). Serum levels of thioredoxin and DJ-1 were significantly higher in NSCLC patients; therefore, these may be utilized as novel diagnostic and prognostic biomarkers for NSCLC.

Modified method combining in situ detoxification with simultaneous saccharification and cofermentation (SSCF) as a single step for converting exploded rice straw into ethanol.

This paper reports an improved fermentation process that includes simultaneous saccharification, detoxification, and cofermentation as steps for producing bioethanol. Rice straw was first steam exploded (SE) or butanone solution exploded (BSE) and then cofermented with Saccharomyces cerevisiae and Candida shehatae. To overcome the inhibitors, the exploded rice straw was continuously and slowly introduced into a 10 L ventilated fermenter. When the fermentation conditions were set to 1.0% initial dry matter, 10% total dry matter, addition rate of 120 mg/min, total fermentation time of 234 h, and dose of 0.1% (NH4)2SO4, yields of 25.8 g/100 g of dry matter ethanol and 88% total sugar use were obtained for BSE rice straw. The ethanol yields were not significantly different between detoxified materials and non-detoxified materials. Most of the furfural, hydroxymethylfurfural (5-HMF), acetic acid, and butanone were removed during the fermentation of non-detoxified materials, and the sugar concentrations were very low. The in situ detoxification and fermentation was effective and inexpensive when the pre-detoxification of exploded materials and the pre-adaptation of strains steps were omitted.

Surgical treatment and prognosis of octogenarians with non-small cell lung cancer.

OBJECTIVE: To explore the clinical characteristics, surgical treatment and prognosis of non-small cell lung cancer (NSCLC) among elderly patients over 80 years. METHODS: The clinical data, surgical methods, perioperative management, postoperative complications and prognosis of 52 NSCLC patients aged over 80 years were retrospectively analyzed. RESULTS: Out of 52 cases, 27 had a long-term smoking history (51.9%) and 44 were with other diseases (84.6%). Lobectomy was done in 32 cases (65.4%), sub-lobectomy in 20 cases (38.5%), including pulmonary wedge resection in 16 cases (30.8%) and lung segment resection in 4 cases (7.7%). The postoperative complication rate was 44.2% (23/52); the complication rate after lobectomy was 62.5% (20/32) and that after sub-lobectomy was 25% (5/20), with significant difference between lobectomy and sub-lobectomy (P<0.05). Postoperative mortality was 3.8% (2/52). Pathological TNM staging: I a 27 cases (51.9%), I b 12 cases (23.1%), II a 8 cases (15.4%), II b 3 cases (5.8%) and III a 2 cases (3.8%). The 1-, 3- and 5-year survival rate after operation was 87.1%, 59.8%, 19.1%. The 1-, 3- and 5-year survival rate was 86.0%, 61.8%, 21.5% in the patients after lobectomy; that was 89.0%, 58.3%, 18.7% in the patients after sub-lobectomy, with no significant difference between two surgical methods (P>0.05). CONCLUSIONS: Octogenarians with NSCLC are often afflicted with comorbidity, so perioperative management is more complex. Strictly adhering to indications, surgery is still an important treatment of NSCLC patients over 80.

[Technical improvement of mechanical suture of bronchial stumps in pneumonectomy].

OBJECTIVE: To prevent or reduce such postoperative complications of pneumonectomy as bronchopleural fistula and promote the curative effect of the surgery by improving the technique for handling the bronchial stumps. METHODS: In 107 patients, bronchial stapler was adopted for closure of the bronchial stump and pneumonectomy was subsequently performed on the side of the staples. A 1-0 polyester suture was used to for suture in the shape of "U" deep in the mechanical suture on the bronchial stump. The postoperative complications were observed. RESULTS: The operations were performed easily and the bronchial stumps were short and sutured firmly. No fluid retention or granuloma occurred in the stumps after the operation, nor were the complications such as intractable cough and infection of thoracic cavity observed. No bronchopleural fistula was found in these patients. CONCLUSION: This new method is safe and effective for treatment of the bronchial stumps.