Enantioselective fate of famoxadone during processing of apple cider and grape wine.

Famoxadone enantiomers were separated on Lux Amylose-1 chiral column and determined by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). The half-lives of R-(-)-famoxadone and S-(+)-famoxadone were 69.3 and 86.6 h in apple cider, 231.0 and 346.5 h in apple pomace, 69.3 and 77.0 h in grape wine, and 231.0 and 346.5 h in grape pomace, respectively. The enantiomeric fraction (EF) values decreased gradually from 0.498, 0.499, and 0.500 (0 h) to 0.404, 0.374, and 0.427 (144 h) and then increased gradually to 0.474, 0.427, and 0.422 (312 h) in apple cider, grape wine, and grape pomace. The EF value in apple pomace decreased gradually from 0.499 (0 h) to 0.450 (168 h) and then increased gradually to 0.482 (312 h). The processing factors (PFs) for famoxadone ranged from 0.014 to 0.024 in the overall process. The residue of famoxadone reduced 94.7-97.4% after the fermentation process.

[Clinical characterization and genetic analysis of a newborn with chromosome 8q21.11 deletion syndrome].

OBJECTIVE: To explore the genetic etiology for a newborn with corneal opacity. METHODS: The neonate and her parents were subjected to routine G-banding chromosomal karyotyping analysis. Copy number variation (CNV) was analyzed with low-coverage whole-genome sequencing (WGS) and single nucleotide polymorphism microarray (SNP array). RESULTS: No karyotypic abnormality was found in the newborn and her parents. Low-coverage WGS has identified a de novo 5.5 Mb microdeletion at chromosome 8q21.11-q21.13 in the neonate, which encompassed the ZFHX4 and PEX2 genes. The result was confirmed by SNP array-based CNV analysis. CONCLUSION: The newborn was diagnosed with chromosome 8q21.11 deletion syndrome. ZFHX4 may be one of the key genes underlying this syndrome.

Optimization of kinetics and operating parameters for the bioleaching of heavy metals from sewage sludge, using co-inoculation of two Acidithiobacillus species.

This study explores the potential for synchronous extraction of Cu, Cr, Ni and Zn during sewage sludge bioleaching processes, using three types of bacterial cultures: a pure culture of Acidithiobacillus ferrooxidans (A. ferrooxidans); a pure culture of Acidithiobacillus thiooxidans (A. thiooxidans); and a mixed culture of A. ferrooxidans and A. thiooxidans. Variable operating parameters included initial pH, solids concentration, sulfur concentration and ferrous iron concentration, with optimization via Box-Behnken design of response surface methodology. Results indicate that the mixed culture of A. ferrooxidans and A. thiooxidans, was the most effective at bioleaching heavy metals from sewage sludge. The optimal operating conditions were as follows: an initial pH of 2.0, with concentrations of 3% solids, 6.14 g L-1 sulfur and 4.55 g L-1 ferrous iron. Maximum extraction efficiencies obtained after 14 days of bioleaching under optimal conditions, were 98.54% Cu, 57.99% Cr, 60.06% Ni and 95.60% Zn. Bioleaching kinetics were effectively simulated using a shrinking core model to explain the leaching reaction, with modelling results suggesting that the rate was determined by the diffusion step.

Determination of mancozeb residue in fruit by derivatization and a modified QuEChERS method using ultraperformance liquid chromatography-tandem mass spectrometry.

A liquid chromatography-tandem mass spectrometry method with derivatization and a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) sample preparation was developed for the determination of mancozeb in fruits. The target compound was determined in less than 4.0 min with use of an electrospray ionization source in positive mode. The limits of detection and the limits of quantification ranged from 1.0 to 3.2 µg kg-1 and from 10 to 15 µg kg-1 in fruit respectively. The linearity was excellent for mancozeb (R 2 ≥ 0.9920). Recoveries in five matrices were obtained at three spiking levels (0.02, 0.1, and 1 mg kg-1). For all concentrations, the mean recoveries ranged from 84.0% to 95.9%, with repeatability relative standard deviation (n = 5) of 0.6 - 7.0%. The interday reproducibility relative standard deviation (n = 3) ranged from 1.4% to 5.5%. This method could be used for the routine detection of mancozeb residues in fruit. Graphical abstract The derivation of mancozeb and the detection process of derivative product by UPLC-MS/MS.

A molecularly imprinted polymer synthesized using ß-cyclodextrin as the monomer for the efficient recognition of forchlorfenuron in fruits.

In this study, we have successfully synthesized a forchlorfenuron (CPPU)-imprinted polymer in dimethyl sulfoxide by molecular imprinting technique. In the molecular imprinting, ß-cyclodextrin was used as the monomer with 1,6-hexamethylene diisocyanate (HMDI) as the cross-linking agent that formed a complex with forchlorfenuron by inclusion interactions. The adsorption equilibrium was attained in approximately 30 min and the maximum binding capacity was 26.79 mg/g, which indicated that the adsorption kinetics was relatively fast. The results of adsorption and selectivity experiments have shown that the imprinted polymer was able to bind forchlorfenuron specifically and reversibly. The specific recognition of the imprinted polymer for forchlorfenuron resulted from the cooperative effects of inclusion interaction and hydrophobic interaction. This imprinted polymer was also used to accurately determine forchlorfenuron residue in real fruit samples. In the standard spiked strawberry sample, the adsorption recoveries for forchlorfenuron were as high as 90.3, 84.5, and 90.8%, respectively. The binding specificity of the imprinted polymer for forchlorfenuron resulted from the imprinting effect. Therefore, as a reusable material possessing high affinity and selectivity, the imprinted polymer is promising in application to detect pesticide residues in fruit samples. In addition, because ß-cyclodextrin is water soluble and can include neutral compounds, this molecularly imprinted polymer can be used to recognize pesticides in aqueous solutions. Graphical abstract Schematic demonstration of molecular imprinting and re-binding of forchlorfenuron.

Clinical value of circulating tumour cells in evaluating the efficacy of continuous hepatic arterial infusion among colorectal cancer patients.

Few studies have been conducted to evaluate the efficacy of HAIC using circulating tumour cells (CTCs). In this study, a total of 100 patients who received HAIC treatment and CTC detection were selected. The results showed that after HAIC treatment, the levels of CTC, carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA) decreased. Postoperative progression-free survival (PFS) rates between patients with positive and negative preoperative CTC results, and for CA19-9, CEA were significantly different. The positive rate of CTCs was 61% before chemotherapy and 23% after chemotherapy, and the correlation coefficient between the two was 0.385. Those whose CTC values increased after chemotherapy had shorter PFS rates. CTCs are an independent predictor of recurrence. Patients with CTC-positive results are more susceptible to recurrence. The CTC count in peripheral blood has a close bearing on the postoperative chemotherapy efficacy of patients with CRC and affects patients' PFS.

The mechanisms of tumor necrosis factor α in regulating Krüpple-like factor 4 expression in SK-BR-3 breast cancer cells.

OBJECTIVE: To explore the expression and functional role of Krüpple-like factor 4 (KLF4) protein stimulated by tumor necrosis factor α (TNF-α) in SK-BR-3 breast cancer cells. METHODS: SK-BR-3 cells were stimulated with various concentrations of TNF-α at 0, 1, 5, 10, and 20 ng/mL. Expression levels of KLF4 protein were detected by Western blotting. In the detection of apoptosis, flow cytometry, and DAPI staining were used for detecting the level of apoptosis. RESULTS: KLF4 expression was markedly elevated following stimulation of SK-BR-3 with TNF-α. At the same time, the expression of KLF4 protein increased gradually with the increase of TNF-α stimulation concentration. TNF-α stimulation of SK-BR-3 cells increased apoptosis as measured by apoptosis levels. By overexpressing KLF4 protein in SK-BR-3 cells, it similarly increased apoptosis and promoted cell death of SK-BR-3 cells. CONCLUSION: TNF-α promotes KLF4 expression, while TNF-α promotes apoptosis in SK-BR-3 cells, a process that may be due to elevated KLF4 protein expression.

Optical image encryption via ptychography.

Ptychography is combined with optical image encryption for the first time. Due to the nature of ptychography, not only is the interferometric optical setup that is usually adopted not required any more, but also the encryption for a complex-valued image is achievable. Considering that the probes overlapping with each other is the crucial factor in ptychography, their complex-amplitude functions can serve as a kind of secret keys that lead to the enlarged key space and the enhanced system security. Further, since only introducing the probes into the input of common system is required, it is convenient to combine ptychography with many existing optical image encryption systems for varied security applications.

Static magnetic field improvement of the quality of rice dumpling subjected to freeze-thaw cycles: Roles of phase transition of water and changes in structural and physicochemical properties of glutinous rice flour.

This study aimed to investigate the effect of static magnetic field (SMF, 0-10 mT) on the quality of rice dumplings subjected to 7, 14, 21, and 28 freeze-thaw cycles. The underlying mechanism was explored by monitoring changes in water phase transition, water distribution, and structural and physicochemical properties of rice flour. Results suggested that SMF enables the formation of small ice crystals by accelerating freezing rate, shortening phase transition time, and increasing bound water content, which attributes to reducing the mechanical damage on starch granules and thus improves the quality of frozen rice dumpling. After 7-28 freeze-thaw cycles, SMF treatment increased the whiteness by 0.08-1.58, reduced the cracking ratio by 1.67 %-8.34 %, decreased the water loss ratio by 0-0.75 %, and significantly improved the texture of cooked rice dumplings. This study confirmed the feasibility of SMF in improving the quality of rice dumpling, which contributes to expanding the applications of magnetic freezer in the preservation of starch-based foods.

Metabolomic and transcriptomic analyses of the flavonoid biosynthetic pathway in blueberry (Vaccinium spp.).

As a highly economic small fruit crop, blueberry is enjoyed by most people in terms of color, taste, and rich nutrition. To better understand its coloring mechanism on the process of ripening, an integrative analysis of the metabolome and transcriptome profiles was performed in three blueberry varieties at three developmental stages. In this study, 41 flavonoid metabolites closely related to the coloring in blueberry samples were analyzed. It turned out that the most differential metabolites in the ripening processes were delphinidin-3-O-arabinoside (dpara), peonidin-3-O-glucoside (pnglu), and delphinidin-3-O-galactoside (dpgal), while the most differential metabolites among different varieties were flavonols. Furthermore, to obtain more accurate and comprehensive transcripts of blueberry during the developmental stages, PacBio and Illumina sequencing technology were combined to obtain the transcriptome of the blueberry variety Misty, for the very first time. Finally, by applying the gene coexpression network analysis, the darkviolet and bisque4 modules related to flavonoid synthesis were determined, and the key genes related to two flavonoid 3', 5'-hydroxylase (F3'5'H) genes in the darkviolet module and one bHLH transcription factor in the bisque4 module were predicted. It is believed that our findings could provide valuable information for the future study on the molecular mechanism of flavonoid metabolites and flavonoid synthesis pathways in blueberries.

Identification of pathogens and detection of antibiotic susceptibility at single-cell resolution by Raman spectroscopy combined with machine learning.

Rapid, accurate, and label-free detection of pathogenic bacteria and antibiotic resistance at single-cell resolution is a technological challenge for clinical diagnosis. Overcoming the cumbersome culture process of pathogenic bacteria and time-consuming antibiotic susceptibility assays will significantly benefit early diagnosis and optimize the use of antibiotics in clinics. Raman spectroscopy can collect molecular fingerprints of pathogenic bacteria in a label-free and culture-independent manner, which is suitable for pathogen diagnosis at single-cell resolution. Here, we report a method based on Raman spectroscopy combined with machine learning to rapidly and accurately identify pathogenic bacteria and detect antibiotic resistance at single-cell resolution. Our results show that the average accuracy of identification of 12 species of common pathogenic bacteria by the machine learning method is 90.73 ± 9.72%. Antibiotic-sensitive and antibiotic-resistant strains of Acinetobacter baumannii isolated from hospital patients were distinguished with 99.92 ± 0.06% accuracy using the machine learning model. Meanwhile, we found that sensitive strains had a higher nucleic acid/protein ratio and antibiotic-resistant strains possessed abundant amide II structures in proteins. This study suggests that Raman spectroscopy is a promising method for rapidly identifying pathogens and detecting their antibiotic susceptibility.

Monitoring Volatile Organic Compounds in Different Pear Cultivars during Storage Using HS-SPME with GC-MS.

Aroma, which plays an essential role in food perception and acceptability, depends on various mixture of volatile organic compounds (VOCs). Meanwhile, as a field of metabolomics, VOC analysis is highly important for aroma improvement and discrimination purposes. In this work, VOCs in pear fruits were determined via headspace solid-phase micro-extraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS) to study variations among different cultivars and storage stages. In 12 cultivars of pear fruits, a total of 121 VOCs were quantified, including 40 esters, 32 alcohols, 16 aldehydes, 13 alkenes, 11 ketones, 4 acids, and 5 other compounds. The types and amounts of VOCs in different cultivars varied dramatically, which were in the range of 13-71 and 3.63-55.65 mg/kg FW (fresh weight), respectively. The Dr. Guyot cultivar showed the highest level of VOCs, both in type and amount. After 21 days storage at 4 °C, total concentration of VOCs increased from initial levels of 50.76 to 101.33 mg/kg FW. Storage at 20 °C made a larger contribution to production for VOCs than that at 4 °C, resulting in the maximum content of VOCs (117.96 mg/kg FW) in fruit after 14 days storage at 4 °C plus 7 days at 20 °C. During storage, the content of esters showed a gradual increase, while the content of alcohols and aldehydes decreased. Based on the results presented, related alcohols were recognized as the intermediates of conversion from aldehydes to esters.

Risk Assessment of Pesticide Residues in Chinese Litchis.

ABSTRACT: The presence of pesticide residues in fruit has been of extensive concern worldwide. Pesticide residues in 150 litchi samples collected in the People's Republic of China were measured, and the dietary exposure risks to consumers were evaluated. The litchi samples were screened by gas chromatography and ultraperformance liquid chromatography with tandem mass spectrometry for the possible presence of 57 pesticides. Seventeen pesticides were detected, and 70.7% of samples contained residues of one or more pesticides. The most frequently detected pesticide (36% of samples) was diflubenzuron. Carbofuran in one sample exceeded its maximum residue limit by 125%. In dietary exposure assessments, all concentrations below the limit of detection (LOD) were calculated as 0, 0.5 × the LOD, or at the LOD separately, and these assessments indicated that the chronic health risks from these dietary exposures were extremely low. For acute exposures, carbofuran was at 11.08% of the acute reference dose, and all other pesticides were <2% of the acute reference dose. The findings indicated that the presence of pesticide residues in litchis should not be considered a public health problem.

[Analysis of GJB2 gene coding sequence in patients with nonsyndromic hearing loss].

OBJECTIVE: To analyze the coding sequence of GJB2 gene in six pedigrees with nonsyndromic hearing loss in order to find deafness-causing mutations in the GJB2 gene, and to explore the inherent pattern of deafness-causing mutations in the GJB2 gene. METHODS: Genomic DNA was extracted from peripheral blood for the probands and their family members. Coding sequence of the GJB2 gene was amplified by polymerase chain reaction, sequence variations were determined by DNA sequencing. Amplified fragments with overlapping peaks on sequencing chromatogram were sequenced by TA cloning in order to determine whether the mutations originated from the same allele. RESULTS: Mutations in the GJB2 gene were found in 4 out of the 6 pedigrees with nonsyndromic hearing loss. Four types of mutations were detected in the GJB2 gene, which were 235delC, 299-300delAT, 79G>A+341A>G, and 109G>A. Compound heterozygous polymorphisms 79G>A and 341A>G, and mutations 109G>A and 235delC had deafness-causing effects. CONCLUSION: Heterogeneous mutations of the GJB2 gene are frequently seen in patients with nonsyndromic hearing loss. Sometimes, polymorphisms may cause deafness when they are combined. Environmental factors and other genes may contribute to hearing loss caused by the GJB2 gene mutations.

Fuzzy C-Means Clustering Algorithm-Based Magnetic Resonance Imaging Image Segmentation for Analyzing the Effect of Edaravone on the Vascular Endothelial Function in Patients with Acute Cerebral Infarction.

This paper aimed to discuss the denoising ability of magnetic resonance imaging (MRI) images based on fuzzy C-means clustering (FCM) algorithm and the influence of Butylphthalide combined with Edaravone treatment on nerve function and vascular endothelial function in patients with acute cerebral infarction (ACI). Based on FCM algorithm, Markov Random Field (MRF) model algorithm was introduced to obtain a novel algorithm (NFCM), which was compared with FCM and MRF algorithm in terms of misclassification rate (MCR) and difference of Kappa index (KI). 90 patients with ACI diagnosed in hospital from December 2018 to December 2019 were selected as subjects, who were divided into combined treatment group (conventional treatment + Edaravone + Butylphthalide) and Edaravone group (conventional treatment + Edaravone) randomly, each consisting of 45 cases. The National Institutes of Health Stroke Scale (NIHSS) score and endothelial function index level such as plasma nitric oxide (NO), human endothelin-1 (ET-1), and vascular endothelial cell growth factor (VEGF) were compared before and after treatment between the two groups. The results showed that the MCR of NFCM was evidently inferior to FCM and MRF, and the KI was notably higher relative to the other two algorithms. After treatment, the NIHSS score of the combined treatment group was (9.09 ± 1.86) points and that of Edaravone group was (14.97 ± 3.44) points, with evident difference between the two groups (P < 0.05). After treatment, the NO of the combined treatment was (54.63 ± 4.85), and that of Edaravone group was (41.54 ± 5.27), which was considerably different (P < 0.01), and the VEGF and ET-1 of combined treatment group were greatly inferior to Edaravone group (P < 0.01). It was revealed that the novel algorithm based on FCM can obtain more favorable quality and segmentation accuracy of MRI images. Moreover, Butylphthalide combined with Edaravone treatment can effectively improve nerve function, vascular endothelial function, and short-term prognosis in ACI, which was safe and worthy of clinical adoption.

DNA sequencing, genomes and genetic markers of microbes on fruits and vegetables.

The development of DNA sequencing technology has provided an effective method for studying foodborne and phytopathogenic microorganisms on fruits and vegetables (F & V). DNA sequencing has successfully proceeded through three generations, including the tens of operating platforms. These advances have significantly promoted microbial whole-genome sequencing (WGS) and DNA polymorphism research. Based on genomic and regional polymorphisms, genetic markers have been widely obtained. These molecular markers are used as targets for PCR or chip analyses to detect microbes at the genetic level. Furthermore, metagenomic analyses conducted by sequencing the hypervariable regions of ribosomal DNA (rDNA) have revealed comprehensive microbial communities in various studies on F & V. This review highlights the basic principles of three generations of DNA sequencing, and summarizes the WGS studies of and available DNA markers for major bacterial foodborne pathogens and phytopathogenic fungi found on F & V. In addition, rDNA sequencing-based bacterial and fungal metagenomics are summarized under three topics. These findings deepen the understanding of DNA sequencing and its application in studies of foodborne and phytopathogenic microbes and shed light on strategies for the monitoring of F & V microbes and quality control.

Ursolic acid ameliorates adipose tissue insulin resistance in aged rats via activating the Akt-glucose transporter 4 signaling pathway and inhibiting inflammation.

Ageing often results in insulin resistance (IR) and chronic inflammation, and adipose is one of the tissues in which inflammation and IR occur earliest during this process. The present study investigated the effect and underlying mechanisms of ursolic acid (UA) on adipose IR and inflammation in ageing rats. Specific pathogen-free male Sprague-Dawley rats were randomly divided into 4 groups: i) Young normal (young); ii) untreated ageing (aged); and groups supplemented with UA either iii) low-UA 10 mg/kg (UA-L) or iv) high-50 mg/kg (UA-H). Animals in the UA-treated groups received 10 or 50 mg/kg UA (suspended in 5% Gum Arabic solution). The rats in the corresponding aged group and young groups received vehicle (5% Gum Arabic) alone. All rats were intragastrically treated once daily by oral gavage for 7 weeks. The day before the experiment terminated, overnight fasting blood (~700 µl) was collected and plasma was prepared to measure biochemical indicators; western blotting was performed to analyze the expression of insulin signaling proteins [(insulin receptor substrate 1 (IRS-1), phosphorylated (p)-IRS-1, PI3K, glucose transporter 4 (GLUT4), Akt and p-Akt)] and inflammatory factors (NF-κB, IL-6 and IL-1ß) in the epididymis white adipose tissue (eWAT). The results revealed that treatment with UA-H decreased eWAT weight, the ratio of eWAT weight/body weight, fasted insulin and triglyceride levels, the homeostasis model assessment of insulin resistance and adipose tissue insulin resistance index in ageing rats, indicating the amelioration of systemic and adipose tissue IR, compared with the aged group. Mechanistically, UA-H administration upregulated p-protein kinase B, the ratio of p-Akt to protein kinase B and total and cellular membrane GLUT4 protein levels in eWAT of ageing rats. Conversely, UA inhibited the increase in NF-κB expression and proinflammatory cytokines IL-6 and IL-1ß. However, these alterations were not observed in the rats of the aged group. Taken together, the findings of the present study indicated that UA may ameliorate adipose IR, which is associated with activation of the Akt-GLUT4 signaling pathway and inhibition of inflammation in ageing rats. These data provide a basis for the development of effective and safe drugs or functional substances, such as UA, for the prevention and treatment of metabolic diseases.

Apple pomace and rosemary extract ameliorates hepatic steatosis in fructose-fed rats: Association with enhancing fatty acid oxidation and suppressing inflammation.

Apple pomace and rosemary (AR) have been reported to contain rich bioactive molecules, which have numerous metabolic effects. Our preliminary work revealed that AR ameliorated fructose-induced insulin resistance in rats by modulating sarcolemmal CD36 and glucose transporter-4. The present study aimed to further examine how AR improves metabolic disorders by investigating the effect of AR on hepatic steatosis induced by fructose overconsumption. The results demonstrated that AR (100 mg/kg daily by gavage for 5 weeks) attenuated chronic liquid fructose consumption-induced increases in liver triglyceride content in rats. Mechanistically, reverse transcription-quantitative PCR and western blot analysis results indicated that AR reversed fructose-induced suppression of hepatic peroxisome proliferator-activated receptor α, carnitine palmitoyl-transferase 1α, sirtuin 1 and peroxisome proliferator-activated receptor-γ coactivator 1α, which were associated with the fatty acid oxidative (FAO) pathway. In addition, AR treatment decreased the expression levels of the pro-inflammatory proteins NF-κB and tumor necrosis factor-α. However, AR had no effect on the genes related to lipogenesis and the very low-density lipoprotein-export pathway in rat liver. Thus, the present results suggested that AR treatment diminished long-term fructose overconsumption-induced fatty liver, which was associated with enhanced FAO and suppressed inflammation.

Construction and Characterization of KRAS Immune Lipid Magnetic Balls for Colorectal Cancer Circulating Tumor Cells.

OBJECTIVE: The purpose of this study was to prepare and characterize a lipid magnetic ball modified with KRAS antibodies on the surface and to isolate circulating tumor cells of colorectal cancer with KRAS mutations. METHODS: The microemulsion method was used to form lipid bilayers to encapsulate Fe3O4 nanoparticles with superparamagnetism to form lipid magnetic balls, and KRAS antibodies were formed on the surface to form KRAS immune lipid magnetic balls. RESULTS: Compared with traditional EpCAM antibody-modified lipid magnetic balls, it can effectively improve the capture ability of colorectal cancer circulating tumor cells with KRAS mutation, the capture rate reaches 92.9%, and the capture results are consistent with clinical diagnosis and pathology. CONCLUSION: Our results showed that KRAS antibody-modified lipid magnetic balls can be used in the diagnosis and treatment of KRAS colorectal cancer.