Quantitative Textural and Rheological Data on Different Levels of Texture-Modified Food and Thickened Liquids Classified Using the International Dysphagia Diet Standardisation Initiative (IDDSI) Guideline.

Diet modification is a common compensation strategy to promote swallowing safety in patients with swallowing difficulties. The International Dysphagia Diet Standardisation Initiative (IDDSI) guideline provides qualitative descriptions on texture-modified food and thickened liquid. This study aimed to establish quantitative textural and rheological data on different IDDSI levels based on common Chinese ingredients and dishes. Textural and rheological properties of 226 samples of various food textures and 93 samples of various liquid consistencies were obtained using a texture profile analysis (TPA) and viscometer, respectively. The establishment of such quantitative data can be used for future texture-modified food product development and research purposes.

Combination of untargeted and targeted proteomics for secretome analysis of L-WRN cells.

Organoid culture is a promising biomedical technology that requires specialized growth factors. Recently, a recombinant L-WRN cell line has been extensively used to generate conditioned medium (L-CM) for organoid culture. Nevertheless, methods for evaluating the stability of the L-WRN cells have been limited. In this study, a novel proteomics-based approach was developed to analyze the secretome of the cells. Serum-free L-CM was lyophilized, precipitated by trichloroacetic acid, and desalted prior to analysis by liquid chromatography-tandem mass spectrometry. Data-dependent acquisition (DDA) was conducted for the untargeted secretome profiling of the cells, and parallel reaction monitoring (PRM) was applied for the targeted quantification of the Wnt3A, R-spondin3, and noggin proteins (WRNs). This study also compared the performance of two types of PRM methods, namely MS1-independent PRM and MS1-dependent PRM, that can be executed on an Orbitrap instrument. The results showed that the growth of mouse intestinal organoids was closely related to the use of L-CM. The composition of L-CM could be markedly affected by the medium collection scheme. A total of 1725, 2302, and 2681 proteins were identified from the L-CM collected on day 5, day 9, and day 13, respectively. The MS1-independent PRM outperformed the MS1-dependent PRM and effectively quantified the WRNs with high repeatability and specificity. In conclusion, by integrating untargeted and targeted proteomics, this study develops a mass spectrometry-based method for the secretome analysis and quality control of the L-WRN cells. The methodology and findings of the present work will benefit future studies on organoids and secretomes.

Pseudo-Taste Cells Derived from Rat Taste and Non-Taste Tissues: Implications for Cultured Taste Cell-Based Biosensors.

Although the technique for taste cell culture has been reported, cultured taste cells have remained poorly validated. This study systematically compared the cultured cells derived from both taste and non-taste tissues. Fourteen cell lines established from rat circumvallate papillae (RCVs* or RCVs), non-taste lingual epithelia (RVEs), and tail skins (RTLs) were analyzed by PCR, immunocytochemistry, proteomics, and calcium imaging. The cell lines were morphologically indistinguishable, and all expressed some taste-related molecules. Of the tested RCVs*, RCVs, RVEs, and RTLs (%), 84.7 ± 7.8, 63.9 ± 22.8, 46.8 ± 0.3, and 40.8 ± 15.1 of them were responsive to at least one tastant or ATP, respectively. However, the calcium signaling pathways in the responding cells differed from the canonical taste transduction pathways in the taste cells in vivo, suggesting that they were not genuine taste cells. In addition, the growth medium intended for taste cell culture did not prevent the proliferation of non-gustatory epithelial cells regardless of supplementation of Y-27632 and EGF. In conclusion, the current method for taste cell culture is susceptible to pseudo-taste cells that may lead to overinterpretation. Thus, biosensors that rely on calcium responses of cultured taste cells should be applied with extreme caution.

Dynamic correlations between major enzymatic activities, physicochemical properties and targeted volatile compounds in naturally fermented plain sufu during production.

Dynamic changes and correlations between physicochemical properties, 14 targeted volatile compounds (TVCs) and six groups of enzyme activities during eight production stages of naturally fermented plain sufu were explored. Multiple factor analysis was used to discriminate between and group the samples into three clusters: cluster I comprised tofu and pehtze; cluster II comprised dried pehtze and salted pehtze; cluster III involved the aging stages. Clusters I and II were characterised by higher enzyme activities, while cluster III was characterised by the presence of diverse TVCs. Protease and esterase were strongly correlated with most of the TVCs. Esterase, in particular, contributed to the formation of three high molecular weight esters, namely, ethyl dodecanoate, ethyl (Z)-9-octadecenoate and ethyl (Z, Z)-9,12-octadecadienoate. The enzymes found contributed to the texture and flavour of naturally fermented sufu and will provide a good guide and control for using the enzymes directly to ripen sufu products.

Saltiness-Enhancing Peptides Isolated from the Chinese Commercial Fermented Soybean Curds with Potential Applications in Salt Reduction.

Salt is very important for human health and food seasoning. Recently, several peptides isolated from natural food products have been reported exhibiting a salty taste or a saltiness-enhancing function. In this investigation, taste-active peptides occurring in commercial Chinese fermented soybean curd were isolated and identified using ultrafiltration, gel permeation chromatography, ion-exchange chromatography, and nano-LC/Q-TOF MS/MS. The salty taste-enhancing function of the target fractions was confirmed by both a rat taste cell model and/or human sensory evaluation. Four decapeptides were found as taste-active compounds. Among them, peptide E (EDEGEQPRPF) was the most potent saltiness-enhancing peptide: 0.4 mg/mL in 50 mmol/L NaCl solution could increase its salty perception equivalent to the salt level of 63 mmol/L NaCl reference solution. The sequence of the peptide has been found in the α'-subunit of ß-conglycinin [Glycine max]. The remaining peptides V (VGPDDDEKSW), DD (DEDEQPRPIP), and DG (DEGEQPRPFP) showed umami and kokumi tastes as well as a weak saltiness-enhancing sensation. These findings suggest that the decapeptide EDEGEQPRPF could be a possible alternative to partially reduce the amount of sodium intake without compromising for saltiness.

Immortalization and Characterization of Rat Lingual Keratinocytes in a High-Calcium and Feeder-Free Culture System Using ROCK Inhibitor Y-27632.

Cultured keratinocytes are desirable models for biological and medical studies. However, primary keratinocytes are difficult to maintain, and there has been little research on lingual keratinocyte culture. Here, we investigated the effect of Y-27632, a Rho kinase (ROCK) inhibitor, on the immortalization and characterization of cultured rat lingual keratinocyte (RLKs). Three Y-27632-supplemented media were screened for the cultivation of RLKs isolated from Sprague-Dawley rats. Phalloidin staining and TUNEL assay were applied to visualize cytoskeleton dynamics and cell apoptosis following Y-27632 removal. Label-free proteomics, RT-PCR, calcium imaging, and cytogenetic studies were conducted to characterize the cultured cells. Results showed that RLKs could be conditionally immortalized in a high-calcium medium in the absence of feeder cells, although they did not exhibit normal karyotypes. The removal of Y-27632 from the culture medium led to reversible cytoskeletal reorganization and nuclear enlargement without triggering apoptosis, and a total of 239 differentially expressed proteins were identified by proteomic analysis. Notably, RLKs derived from the non-taste epithelium expressed some molecular markers characteristic of taste bud cells, yet calcium imaging revealed that they rarely responded to tastants. Collectively, we established a high-calcium and feeder-free culture method for the long-term maintenance of RLKs. Our results shed some new light on the immortalization and differentiation of lingual keratinocytes.

Cyperenoic acid, a sesquiterpene derivative from Croton crassifolius, inhibits tumor growth through anti-angiogenesis by attenuating VEGFR2 signal pathway in breast cancer.

BACKGROUND: Cyperenoic acid, one of the main chemical constituents of the root of Croton crassifolius, exhibited potent anti-angiogenic property on the zebrafish embryo model with little cytotoxicity. Nevertheless, its anti-angiogenic mechanism and anti-tumor effect have not been investigated. PURPOSE: To investigate the anti-angiogenic mechanisms of cyperenoic acid and evaluate it whether could exert anti-tumor effect by inhibiting angiogenesis. STUDY DESIGN: Targeting vascular endothelial growth factor receptor-2 (VEGFR2) pathway to inhibit tumor angiogenesis is a significant strategy for cancer treatment. Initially, the anti-angiogenic effect of cyperenoic acid as well as the mechanisms of the action was studied using both in-vitro and in-vivo methodologies. Then, its anti-tumor effect through anti-angiogenesis by attenuating VEGFR2 signaling pathway was evaluated. METHODS: The in-vitro inhibitory effect of cyperenoic acid on the vascular endothelial growth factor (VEGF)-induced angiogenesis was evaluated using human umbilical vein endothelial cells (HUVECs) model. Moreover, its ex-vivo and in-vivo effects were evaluated using the aortic ring assay and the matrigel plug assay. The influence of the cyperenoic acid on tyrosine phosphorylation of VEGFR2 was studied by western blotting assay and the influence on downstream signaling pathway of VEGFR2 also be detected. Computer-docking simulations were carried out to study the interaction between cyperenoic acid and VEGFR2. Finally, its inhibitory effect on tumor growth was studied using breast cancer xenograft model. RESULTS: Cyperenoic acid possessed little toxicity to HUVECs, but it significantly inhibited VEGF-induced proliferation, invasion, migration and tube formation of HUVECs. Moreover, it inhibited VEGF-induced sprout formation ex vivo and vessel formation in vivo. Further mechanistic study showed that cyperenoic acid could suppress VEGFR2 tyrosine kinase activity and alter its downstream signaling pathways in VEGF-induced HUVECs. In addition, it could form two hydrogen bonds with the ATP binding pocket of the VEGFR2 kinase domain by docking. For breast cancer xenograft model, cyperenoic acid suppressed tumor growth, but no obvious toxic pathologic changes were observed in mice. Besides, it suppressed the phosphorylation of VEGFR2 in tumor, demonstrating its anti-angiogenic ability in vivo partly targeting the VEGFR2. CONLUSION: Cyperenoic acid could exert anti-tumor effect in breast cancer by inhibiting angiogenesis via VEGFR2 signaling pathway.

Exploring core functional microbiota related with flavor compounds involved in the fermentation of a natural fermented plain sufu (Chinese fermented soybean curd).

This study aimed to explore the core functional microbiotas related to flavor compounds involving in a naturally fermented soybean curd (plain sufu). Properties such as physicochemical parameters, flavor compounds (17 free amino acids, 21 fatty acids, and 14 aroma volatiles) and microbiota profiles were investigated, and their correlations were explored at 8 stages during production. Results from principal component analysis, multiple factor analysis, and partial least squares-discrimination analysis showed that these properties varied significantly in the eight stages. Furthermore, based on Pearson correlation coefficients and Variable importance for predictive components values between the microbiota profiles and flavor compounds, nine bacterial (Bacillus, Enterobacter, Lactobacillus, Sphingobacterium, Stenotrophomonas, Tetragenococcus, Trabulsiella, Unclassified, and Weissella) and six fungal (Alternaria, Sterigmatomyces, Actinomucor, Fusarium, Debaryomyces, Candida) genera were identified as core functional microbiotas significantly affecting the production of flavor compounds during the natural production. Overall, this study provided a comprehensive description of the dynamic changes of physicochemical parameters, flavor compounds, and microbiota profiles throughout the natural production of plain sufu. The similarities and variations among different stages, as well as correlation between flavor compounds and microbiotas would help to understand the mechanism of plain sufu production, and further to enhance the quality control of plain sufu.

The cholesterol-lowering activity of miracle fruit (Synsepalum dulcificum).

Miracle fruit (Synsepalum dulcificum) is famous for its uniqueness of modifying sour taste to sweetness. However, its cholesterol-lowering activity has not been reported. This study investigated the effect of S. dulcificum on the compositional changes of plasma lipids in hamsters fed a high-cholesterol control diet. Six groups of hamsters were fed either a control diet or one of the five experimental diets containing 2% ethanol extract of leaves, 2% water extract of leaves, 2% ethanolic extract of seeds (ES), 2% water extract of seeds, or 2% dry pulp. Results showed that ES decreased the plasma total cholesterol (TC). Two triterpenoids (lupeol acetate and ß-amyrin acetate) were isolated from the ES and they added to a diet could decrease TC by 15%-20% in hamsters. It was concluded that ES showed potent TC-lowering activity and triterpenoid was one of the active components of ES. PRACTICAL APPLICATIONS: In recent years, people are more interested in phytochemicals from functional foods treated for hyperlipidemia because they possessed fewer side effects than the synthetic drugs. The triterpenoids isolated from the miracle fruit may be promising candidates for the development of cholesterol-lowering agent, especially for patients whose blood cholesterol level and body weight are high. Meanwhile, the miracle fruit have a good potential as cholesterol-lowering functional food or a natural source of cholesterol-lowering agent.

Anti-Tumor Activity of Atractylenolide I in Human Colon Adenocarcinoma In Vitro.

Atractylodes macrocephala is known to exhibit multi-arrays of biologic activity in vitro. However, detail of its anti-tumor activity is lacking. In this study, the effects of atractylenolide I (AT-I), a bio-active compound present in Atractylodes macrocephala rhizome was studied in the human colorectal adenocarcinoma cell line HT-29. The results showed that AT-I induced apoptosis of human colon cancer cells through activation of the mitochondria-dependent pathway. The IC50 of AT-I was 277.6 µM, 95.7 µM and 57.4 µM, after 24, 48 and 72 h of incubation with HT-29, respectively. TUNEL and Annexin V-FITC/PI double stain assays showed HT-29 DNA fragmentation after cell treatment with various AT-I concentrations. Western blotting analysis revealed activation of both initiator and executioner caspases, including caspase 3, caspase 7, and caspase 9, as well as PARP, after HT-29 treatment with AT-I via downregulation of pro-survival Bcl-2, and upregulation of anti-survival Bcl-2 family proteins, including Bax, Bak, Bad, Bim, Bid and Puma. The studies show for the first time that AT-I is an effective drug candidate towards the HT-29 cell.

Multivariate relationships among sensory, physicochemical parameters, and targeted volatile compounds in commercial red sufus (Chinese fermented soybean curd): Comparison of QDA® and Flash Profile methods.

Selected physico- (texture and color) chemical (salt, protein, and moisture content) properties and 20 targeted volatile compounds (TVCs) from 12 commercial red sufus were investigated to identify correlations with sensory attributes obtained by either QDA or Flash Profile (FP). The intensities of 15 attributes from QDA and the ranking values of 19 attributes from FP of red sufus were evaluated, and a higher variance was generally found in the results from FP than those from QDA among the 12 samples. Principal component analysis (PCA) of TVCs showed that both the concentration and the ratio of volatile compounds influenced the flavor quality and the discrimination among the samples. Multiple factor analysis (MFA) was used to associate the sensory data from either FP or QDA with the physicochemical properties and TVCs of red sufus. The results showed that both FP and QDA data associated well with the physicochemical and TVC data, and the intensity of the sensory attributes could be predicted from the properties of red sufus. Pearson correlation coefficients between the sensory attributes (aroma/flavor) and TVCs pinpointed that a sulfur-like aroma was a key attribute in red sufus, and it might be a result of the comprehensive combination of different TVCs. The information reported here could be important for the quality control of traditional and new variants of red sufu products by providing an approach to substitute the sensory measurements with the instrumental measurements, and to strengthen the interpretation of sensory data by showing how they are affected by the physicochemical properties.

Quantitative Structure-Activity Relationship Study of Bitter Di-, Tri- and Tetrapeptides Using Integrated Descriptors.

New quantitative structure-activity relationship (QSAR) models for bitter peptides were built with integrated amino acid descriptors. Datasets contained 48 dipeptides, 52 tripeptides and 23 tetrapeptides with their reported bitter taste thresholds. Independent variables consisted of 14 amino acid descriptor sets. A bootstrapping soft shrinkage approach was utilized for variable selection. The importance of a variable was evaluated by both variable selecting frequency and standardized regression coefficient. Results indicated model qualities for di-, tri- and tetrapeptides with R2 and Q2 at 0.950 ± 0.002, 0.941 ± 0.001; 0.770 ± 0.006, 0.742 ± 0.004; and 0.972 ± 0.002, 0.956 ± 0.002, respectively. The hydrophobic C-terminal amino acid was the key determinant for bitterness in dipeptides, followed by the contribution of bulky hydrophobic N-terminal amino acids. For tripeptides, hydrophobicity of C-terminal amino acids and the electronic properties of the amino acids at the second position were important. For tetrapeptides, bulky hydrophobic amino acids at N-terminus, hydrophobicity and partial specific volume of amino acids at the second position, and the electronic properties of amino acids of the remaining two positions were critical. In summary, this study not only constructs reliable models for predicting the bitterness in different groups of peptides, but also facilitates better understanding of their structure-bitterness relationships and provides insights for their future studies.

Optimization of a headspace solid-phase micro-extraction method to quantify volatile compounds in plain sufu, and application of the method in sample discrimination.

A rapid headspace solid-phase microextraction method was optimized to detect volatile components in three commercial plain sufu samples. The method was then used to qualify 14 aroma impact components that contribute significantly to the volatile profile in 12 commercial samples produced at three locations and laboratory-scale fermented samples aged for 3, 15, 30, and 90 days. Principal component analysis (PCA) was subsequently used to group the samples. The optimized method identified 148 volatile compounds in three commercial samples, and the concentrations of 14 aroma impact compounds varied significantly among commercial and laboratory samples. PCA confirmed that these samples could be discriminated according to their production location and aging time, thereby rendering this method a simple strategy for sample discrimination.

Comparison between quantitative descriptive analysis and flash profile in profiling the sensory properties of commercial red sufu (Chinese fermented soybean curd).

BACKGROUND: Red sufu is a type of sufu produced by solid-state fermentation of soybean curd and coloration with red mold rice. The purposes of this study were: (i) to characterize commercial red sufu samples using the quantitative descriptive analysis (QDA) and flash profile (FP) by ten trained and ten untrained panelists, respectively; (ii) to compare the differences in panel performance, descriptive abilities and sensory maps between the two methodologies; and (iii) to compare the efficiency between QDA and FP using red sufu as the matrix. Techniques in multivariate analysis were utilized to explore the data. RESULTS: Results from generalized procrustes analysis (GPA) showed that panel performance by QDA was more repeatable and reached higher homogeneity than that by FP. Despite the confidence ellipse results of the 12 red sufus being better discriminated by QDA, the RV coefficient was high (RV = 0.852) between the configurations of the two-dimensional model (F1 and F2) of the two methodologies, indicating that the two methods are similar and closely related. Overall, QDA provided more accurate and detailed information, while FP provided a similar sensory map on product location and descriptive results. CONCLUSION: The FP technique appeared to be an efficient alternative approach to quickly evaluate sensory properties, including appearance, flavor, aroma and textural properties of an array of red sufu products. © 2018 Society of Chemical Industry.

Application of the ideal profile method to identify an ideal sufu for nonregular consumers.

BACKGROUND: Sufu is a fermented soybean food with unique odor, flavor, and texture, which might not be appreciated by nonregular consumers. To identify the attributes that nonregular consumers appreciated, 12 commercial products bought from a Hong Kong market were tested by 113 consumers according to the ideal profile method (IPM), and rated on 22 attributes. RESULTS: Data provided from nonregular consumers were consistent at both panel and consumer levels. The predicted ideal profile received a higher hedonic score (7.0) than the tested products (6.0). Compared with commercial samples, the ideal sufu has low intensity in the attributes of fermented (aroma and flavor), moldy (aroma), and hard (texture), but high intensity in the soybean-like (aroma and flavor) and sesame oil-like (aroma and flavor) attributes. CONCLUSION: IPM could be used as an alternative approach to collect sensory profiles and preference data directly from consumers for product characterization. It could also be applied to identify the attributes that nonregular consumers appreciated. © 2018 Society of Chemical Industry.

Antioxidant activities of ginger extract and its constituents toward lipids.

Lipid oxidation-a major cause of food product deterioration-necessitates the use of food additives to inhibit food oxidation. Ginger extract (GE) has been reported to possess antioxidant properties. However, components isolated from ginger have been rarely reported to inhibit fat oxidation. Herein, antioxidant properties of GE and four pure components derived from it (6-gingerol, 8-gingerol, 10-gingerol, and 6-shogaol) were examined and their properties were compared to those of butylated hydroxytoluene. GE and the constituent components exhibited antioxidant properties that might be attributed to their hydroxyl groups and suitable solubilizing side chains. 6-Shogaol and 10-gingerol exhibited higher activity at 60°C than 6-gingerol and 8-gingerol. Low antioxidant activity was detected at high temperatures (120/180°C). Overall, GE displayed the strongest dose-dependent antioxidant properties, especially at high temperatures, thereby demonstrating that GE can be employed as a natural antioxidant in lipid-containing processed foods.

Arginyl dipeptides increase the frequency of NaCl-elicited responses via epithelial sodium channel alpha and delta subunits in cultured human fungiform taste papillae cells.

Salty taste is one of the five basic tastes and is often elicited by NaCl. Because excess sodium intake is associated with many health problems, it could be useful to have salt taste enhancers that are not sodium based. In this study, the regulation of NaCl-induced responses was investigated in cultured human fungiform taste papillae (HBO) cells with five arginyl dipeptides: Ala-Arg (AR), Arg-Ala (RA), Arg-Pro (RP), Arg-Glu (RE), and Glu-Arg (ER); and two non-arginyl dipeptides: Asp-Asp (DD) and Glu-Asp (ED). AR, RA, and RP significantly increased the number of cell responses to NaCl, whereas no effect was observed with RE, ER, DD, or ED. We also found no effects with alanine, arginine, or a mixture of both amino acids. Pharmacological studies showed that AR significantly increased responses of amiloride-sensitive but not amiloride-insensitive cells. In studies using small interfering RNAs (siRNAs), responses to AR were significantly decreased in cells transfected with siRNAs against epithelial sodium channel ENaCα or ENaCδ compared to untransfected cells. AR dramatically increased NaCl-elicited responses in cells transfected with NHE1 siRNA but not in those transfected with ENaCα or ENaCδ siRNAs. Altogether, AR increased responses of amiloride-sensitive cells required ENaCα and ENaCδ.

Two natural eudesmane-type sesquiterpenes from Laggera alata inhibit angiogenesis and suppress breast cancer cell migration through VEGF- and Angiopoietin 2-mediated signaling pathways.

Eudesmane-type sesquiterpenes are natural sesquiterpenes with anti-inflammatory properties, but their anti-angiogenic activities are not known. The present study demonstrated that 5α-hydroxycostic acid and hydroxyisocostic acid, two eudesmane-type sesquiterpenes (ETSs), isolated from the herb Laggera alata, possessed anti-angiogenic effects. Under non-toxic dosage, ETSs suppressed VEGF­induced proliferation in human umbilical vein endothelial cells (HUVECs) and vessel formation in zebrafish embryos. Moreover, ETSs inhibited VEGF-stimulated HUVEC migration, stress fibers and tube formation. Results from real­time PCR analysis involving in vivo and in vitro experiments indicated that pro-angiogenic-related mRNA levels were downregulated, including VEGFA, VEGFR2 and Tie2 genes after ETS treatments. Western blot analysis showed that ETSs suppressed VEGF-stimulated VEGFR2 phosphorylation and activation of its downstream molecules, such as Src/AKT/eNOS, FAK, PLCγ/ERK1/2 and p38. Moreover, the VEGF-stimulation of angiopoietin 2 (Ang2) mRNA level increase was significantly downregulated in the presence of ETSs. ETSs inhibited Ang2-induced phosphorylation of the receptor Tie2 in HUVECs, which indicated that ETSs not just suppressed VEGF/VEGFR2 axis, but also the Ang2/Tie2 one. Furthermore, the wound-healing assay revealed that ETSs reduced the migration of Ang2-stimulated human breast cancer (MCF-7) cells. Mechanistically, the anti-migration effect of ETSs correlated with the blockade of Ang2-induced E-cadherin loss and AKT activation. Collectively, the present study suggests that ETSs possess anti-angiogenic ability by interfering the VEGF- and Ang2-related pathways, and they may be good drug candidates.

Use of a least absolute shrinkage and selection operator (LASSO) model to selected ion flow tube mass spectrometry (SIFT-MS) analysis of exhaled breath to predict the efficacy of dialysis: a pilot study.

Selected ion flow tube-mass spectrometry (SIFT-MS) provides rapid, non-invasive measurements of a full-mass scan of volatile compounds in exhaled breath. Although various studies have suggested that breath metabolites may be indicators of human disease status, many of these studies have included few breath samples and large numbers of compounds, limiting their power to detect significant metabolites. This study employed a least absolute shrinkage and selective operator (LASSO) approach to SIFT-MS data of breath samples to preliminarily evaluate the ability of exhaled breath findings to monitor the efficacy of dialysis in hemodialysis patients. A process of model building and validation showed that blood creatinine and urea concentrations could be accurately predicted by LASSO-selected masses. Using various precursors, the LASSO models were able to predict creatinine and urea concentrations with high adjusted R-square (>80%) values. The correlation between actual concentrations and concentrations predicted by the LASSO model (using precursor H3O+) was high (Pearson correlation coefficient = 0.96). Moreover, use of full mass scan data provided a better prediction than compounds from selected ion mode. These findings warrant further investigations in larger patient cohorts. By employing a more powerful statistical approach to predict disease outcomes, breath analysis using SIFT-MS technology could be applicable in future to daily medical diagnoses.

Anti-angiogenic Activity and Mechanism of Sesquiterpene Lactones from Centipeda minima.

Centipeda minima is a Chinese herbal medicine used in the treatment of various diseases including cancer. An ethanol extract of the herb, its four fractions with different polarities, and two volatile oils prepared by steam distillation (SD) and supercritical fluid extraction (SFE) were investigated for their anti-angiogenic activity in a wild-type zebrafish model using a quantitative endogenous alkaline phosphatase (EAP) assay. The SFE oil displayed potent anti-angiogenic activity. Fifteen sesquiterpene lactones (SLs; compounds 1-15) isolated from the SFE oil were evaluated for their anti-angiogenic effect. Results revealed that pseudoguaianolide type SLs (1-8) inhibited vessel formation in the zebrafish embryos while guaianolide type SLs (9-15) showed little effect. Among the active ones, 6-O-angeloylenolin (1), a major component of SFE oil, possessed the strongest effect by reducing vessel formation in zebrafish embryos to 40% of the control value at 29.7 µM. Further study using the Tg (fli1a:EGFP) y1-type zebrafish model revealed that it blocked both intersegmental blood vessels (ISVs) and subintestinal vessels plexus (SIVs) formation in zebrafish embryos. Real-time polymerase chain reaction assay on the wild-type zebrafish embryos suggested that 6-O-angeloylenolin affected multiple molecular targets related to angiogenesis including VEGF receptor, angiopoietin, and its receptors. Taken together, our findings demonstrate that C. minima possesses anti-angiogenic activity, and 6-O-angeloylenolin is a promising candidate for the development of an anti-angiogenic agent.