Combined Metabolome and Transcriptome Analyses Unveil the Molecular Mechanisms of Fruit Acidity Variation in Litchi (Litchi chinensis Sonn.).

Fruit acidity determines the organoleptic quality and nutritive value of most fruits. In litchi, although the organic acid composition of pulps is known, the molecular mechanisms and genes underlying variation in fruit acidity remain elusive. Herein, developing pulps of two contrasting litchi varieties, Huaizhi (HZ, low-acidity) and Boye_No.8 (B8, high-acidity), were subjected to metabolomics and transcriptomics, and the dynamic metabolome and transcriptional changes were determined. Measurements revealed that the dominant acidity-related organic acid in litchi pulps is malate, followed in low levels by citrate and tartrate. Variation in litchi pulps' acidity is mainly associated with significant differences in malate and citrate metabolisms during fruit development. Malic acid content decreased by 91.43% and 72.28% during fruit ripening in HZ and B8, respectively. The content of citric acid increased significantly in B8, while in HZ it was reduced considerably. Differentially accumulated metabolites and differentially expressed genes analyses unveiled fumarate, succinate, 2-oxoglutarate, GABA (γ-aminobutyric acid), phosphoenolpyruvate, and citrate metabolisms as the key driving pathways of litchi fruits' acidity variation. The drastic malate and citrate degradation in HZ was linked to higher induction of fumarate and GABA biosynthesis, respectively. Thirty candidate genes, including three key genes (LITCHI026501.m2, fumarase; LITCHI020148.m5, glutamate decarboxylase; and LITCHI003343.m3, glutamate dehydrogenase), were identified for functional studies toward genetic modulation of litchi fruit acidity. Our findings provide insights into the molecular basis of acidity variation in litchi and provide valuable resources for fruit quality improvement.

A phase I study of the VEGFR kinase inhibitor vatalanib in combination with the mTOR inhibitor, everolimus, in patients with advanced solid tumors.

Purpose Combining small-molecule inhibitors of different targets was shown to be synergistic in preclinical studies. Testing this concept in clinical trials is, however, daunting due to challenges in toxicity management and efficacy assessment. This study attempted to evaluate the safety and efficacy of vatalanib plus everolimus in patients with advanced solid tumors and explore the utility of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) studies as a predictive biomarker. Patients and Methods This single-center, phase I trial containing 70 evaluable patients consisted of a dose escalation proportion based on the traditional "3 + 3" design (cohort IA and IB) and a dose expansion proportion (cohort IIA and IIB). Toxicity was evaluated using the Common Terminology Criteria of Adverse Events. Antitumor activity was assessed using the Modified Response Evaluation Criteria in Solid Tumors. Results The maximum tolerated doses were determined to be vatalanib 1250 mg once daily or 750 mg twice daily in combination with everolimus 10 mg once daily. No treatment-related death occurred. The most common toxicities were hypertriglyceridemia, hypercholesterolemia, fatigue, vomiting, nausea and diarrhea. There was no complete response. Nine patients (12.9%) had partial response (PR) and 41 (58.6%) had stable disease (SD). Significant antitumor activity was observed in neuroendocrine tumors with a disease-control rate (PR + SD) of 66.7% and other tumor types including renal cancer, melanoma, and non-small-cell lung cancer. Conclusions The combination of vatalanib and everolimus demonstrated reasonable toxicity and clinical activity. Future studies combining targeted therapies and incorporating biomarker analysis are warranted based on this phase I trial.

Phase 1 trial of Vismodegib and Erlotinib combination in metastatic pancreatic cancer.

BACKGROUND/OBJECTIVES: Interplay between the Hedgehog (HH) and epidermal growth factor receptor (EGFR) pathways modulating the outcome of their signaling activity have been reported in various cancers including pancreatic ductal adenocarcinoma (PDAC). Therefore, simultaneous targeting of these pathways may be clinically beneficial. This Phase I study combined HH and EGFR inhibition in metastatic PDAC patients. METHODS: Combined effects of HH and EGFR inhibition using Vismodegib and Erlotinib with or without gemcitabine in metastatic solid tumors were assessed by CT. Another cohort of patients with metastatic PDAC was evaluated by FDG-PET and tumor biopsies-derived biomarkers. RESULTS: Treatment was well tolerated with the maximum tolerated dose cohort experiencing no grade 4 toxicities though 25% experienced grade 3 adverse effects. Recommended phase II dose of Vismodegib and Erlotinib were each 150 mg daily. No tumor responses were observed although 16 patients achieved stable disease for 2-7 cycles. Paired biopsy analysis before and after first cycle of therapy in PDAC patients showed reduced GLI1 mRNA, phospho-GLI1 and associated HH target genes in all cases. However, only half of the cases showed reduced levels of desmoplasia or changes in fibroblast markers. Most patients had decreased phospho-EGFR levels. CONCLUSIONS: Vismodegib and Erlotinib combination was well-tolerated although overall outcome in patients with metastatic PDAC was not significantly impacted by combination treatment. Biomarker analysis suggests direct targets inhibition without significantly affecting the stromal compartment. These findings conflict with pre-clinical mouse models, and thus warrant further investigation into how upstream inhibition of these pathways is circumvented in PDAC.

Molecular cloning and functional characterization of AcGST1, an anthocyanin-related glutathione S-transferase gene in kiwifruit (Actinidia chinensis).

KEY MESSAGE: AcGST1, an anthocyanin-related GST, may functions as a carrier to transport anthocyanins from ER to tonoplast in kiwifruit. It was positively regulated by AcMYBF110 through directly binding to its promoter. Anthocyanins are synthesized in the cytoplasmic surface of the endoplasmic reticulum but accumulate predominantly in the vacuole. Previous studies in model and ornamental plants have suggested that a member of the glutathione S-transferase (GST) gene family is involved in sequestration of anthocyanins into the vacuole. However, little is known about anthocyanin-related GST protein in kiwifruit. Here, four putative AcGSTs were identified from the genome of the red-fleshed Actinidia chinensis cv 'Hongyang'. Expression analyses reveal only the expression of AcGST1 was highly consistent with anthocyanin accumulation. Molecular complementation of Arabidopsis tt19 demonstrates AcGST1 can complement the anthocyanin-less phenotype of tt19. Transient expression in Actinidia arguta fruits further confirms that AcGST1 is functional in anthocyanin accumulation in kiwifruit. In vitro assays show the recombinant AcGST1 increases the water solubility of cyanidin-3-O-galactoside (C3Gal) and cyanidin-3-O-xylo-galactoside (C3XG). We further show that AcGST1 protein is localized not only in the ER but also on the tonoplast, indicating AcGST1 (like AtTT19) may functions as a carrier protein to transport anthocyanins to the tonoplast in kiwifruit. Moreover, the promoter of AcGST1 can be activated by AcMYBF110, based on results from transient dual-luciferase assays and yeast one-hybrid assays. EMSAs show that AcMYBF110 binds directly to CAGTTG and CCGTTG motifs in the AcGST1 promoter. These results indicate that AcMYBF110 plays an important role in transcriptional regulation of AcGST1 and, therefore, in controlling accumulation of anthocyanins in kiwifruit.

Biochemical and functional characterization of AcUFGT3a, a galactosyltransferase involved in anthocyanin biosynthesis in the red-fleshed kiwifruit (Actinidia chinensis).

Much of the diversity of anthocyanin pigmentation in plant tissues is due to the action of glycosyltransferases, which attach sugar moieties to the anthocyanin aglycone. This step can increase both their solubility and stability. We investigated the pigmentation of the outer and inner pericarps of developing fruits of the red-fleshed kiwifruit Actinidia chinensis cv. 'Hongyang'. The results show that the red color of the inner pericarp is due to anthocyanin. Based on expression analyses of structural genes, AcUFGT was shown to be the key gene involved in the anthocyanin biosynthetic pathway. Expression of AcUFGT in developing fruit paralleled changes in anthocyanin concentration. Thirteen putative UFGT genes, including different transcripts, were identified in the genome of 'Hongyang'. Among these, only the expression of AcUFGT3a was found to be highly consistent with anthocyanin accumulation. Fruit infiltrated with virus-induced gene silencing showed delayed red colorations, lower anthocyanin contents and lower expressions of AcUFGT3a. At the same time, transient overexpression of AcUFGT3a in both Actinidia arguta and green apple fruit resulted in higher anthocyanin contents and deeper red coloration. In vitro biochemical assays revealed that recombinant AcUFGT3a recognized only anthocyanidins as substrate but not flavonols. Also, UDP-galactose was used preferentially as the sugar donor. These results indicate AcUFGT3a is the key enzyme regulating anthocyanin accumulation in red-fleshed kiwifruit.

Phase I trial of everolimus, gemcitabine and cisplatin in patients with solid tumors.

BACKGROUND: A Phase I trial of the 2-drug regimen of everolimus plus gemcitabine (Cohort I) and the 3-drug regimen of everolimus plus gemcitabine and cisplatin (Cohort II) was performed to determine the maximally tolerated dose (MTD) of both combinations. An expansion cohort (Cohort III) of patients with cholangiocarcinoma or gallbladder carcinoma was treated at the MTD. METHODS: A standard 3 + 3 design dose escalation was used. Everolimus was given on Monday/Wednesday/Friday or daily depending upon the dose level. Gemcitabine and cisplatin were administered on days 1 and 8 of each 21 day cycle. RESULTS: Twelve patients were entered in Cohort I and 15 in Cohort II. The MTD for Cohort I was everolimus 5 mg on Monday/Wednesday/Friday and gemcitabine 800 mg/m(2). For Cohort II, it was everolimus 5 mg on Monday/Wednesday/Friday, gemcitabine 600 mg/m(2), and cisplatin 12.5 mg/m(2). All DLTs in this study were hematologic. Complete responses (CR) were seen in a patient with primary peritoneal carcinoma and another with recurrent pancreatic cancer. Partial responses (PR) were seen in 3 patients: breast, ampullary carcinoma and pheochromocytoma. Of 10 patients enrolled in Cohort III, six patients had stable disease and 4 had progressive disease. CONCLUSIONS: This Phase I clinical trial has demonstrated that these 2-drug and 3-drug combinations are generally well tolerated and safely administered. The main DLTs in both regimens were hematologic, specifically thrombocytopenia. The 3-drug combination can be considered as a platform for future studies in specific tumor types.

Effect of modified atmosphere package on attributes of sweet bamboo shoots after harvest.

Tender bamboo shoots undergo rapid senescence that influences their quality and commercial value after harvest. In this study, the tender sweet bamboo shoots ('Wensun') were packed by a passive modified atmosphere packaging (PMAP) to inhibit the senescence process, taking polyethylene package as control. The increase in CO2 and the decrease in O2 gas concentrations in the headspace atmosphere of the packages were remarkably modified by PMAP treatments. The modified gas atmosphere packaging inhibited the changes in firmness, as well as the content of cellulose, total pectin, and lignin in the cell walls of bamboo shoots. The enzymatic activities of cellulase, pectinase, and polygalacturonase that act on cell wall polysaccharides, and phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, peroxidase, and laccase regulating the lignin biosynthesis were modified by PMAP treatment different from control during storage. The expression levels of the lignin biosynthesis genes PePAL3/4, PeCAD, Pe4CL5, PeC4H, PeCCOAOMT, PeCOMT, cellulose synthase PeCESA1, and related transcription factors PeSND2, PeKNAT7, PeMYB20, PeMYB63, and PeMYB85 were clearly regulated. These results suggest that PMAP efficiently retards the changes in lignin and cell wall polysaccharides, thus delaying the senescence of tender sweet bamboo shoots during storage.

Efficient and Ultrastable Seawater Electrolysis at Industrial Current Density with Strong Metal-Support Interaction and Dual Cl--Repelling Layers.

Direct seawater electrolysis is emerging as a promising renewable energy technology for large-scale hydrogen generation. The development of Os-Ni4Mo/MoO2 micropillar arrays with strong metal-support interaction (MSI) as a bifunctional electrocatalyst for seawater electrolysis is reported. The micropillar structure enhances electron and mass transfer, extending catalytic reaction steps and improving seawater electrolysis efficiency. Theoretical and experimental studies demonstrate that the strong MSI between Os and Ni4Mo/MoO2 optimizes the surface electronic structure of the catalyst, reducing the reaction barrier and thereby improving catalytic activity. Importantly, for the first time, a dual Cl- repelling layer is constructed by electrostatic force to safeguard active sites against Cl- attack during seawater oxidation. This includes a strong Os─Cl adsorption and an in situ-formed MoO4 2- layer. As a result, the Os-Ni4Mo/MoO2 catalyst exhibits an ultralow overpotential of 113 and 336 mV to reach 500 mA cm-2 for HER and OER in natural seawater from the South China Sea (without purification, with 1 m KOH added). Notably, it demonstrates superior stability, degrading only 0.37 µV h-1 after 2500 h of seawater oxidation, significantly surpassing the technical target of 1.0 µV h-1 set by the United States Department of Energy.

Phase I dose escalation study of the PKCι inhibitor aurothiomalate for advanced non-small-cell lung cancer, ovarian cancer, and pancreatic cancer.

Protein kinase C iota (PKCι) is overexpressed in non-small-cell lung cancer, ovarian, and pancreatic cancers, where it plays a critical role in oncogenesis. The gold compound aurothiomalate (ATM) has been shown to inhibit PKCι signaling and exerts potent antitumor activity in preclinical models. We sought to determine the maximum tolerated dose (MTD) of ATM. We conducted a phase I dose escalation trial of ATM in patients with non-small-cell lung cancer, ovarian or pancreatic cancer. Patients received ATM intramuscularly weekly for three cycles (cycle duration 4 weeks) at 25, 50, or 75 mg in a 3+3 design. The dose was not escalated for individual patients. Blood samples were analyzed for elemental gold levels. Patients were evaluated every 4 weeks for toxicity and every 8 weeks for response. Fifteen patients were enrolled in this study. Six patients were treated at 25 mg, seven at 50 mg, and two at 75 mg. There was one dose-limiting toxicity at 25 mg (hypokalemia), one at 50 mg (urinary tract infection), and none at 75 mg. There were three grade 3 hematologic toxicities. The recommended MTD of ATM is 50 mg. Patients received treatment for a median of two cycles (range 1-3). There appeared to be a dose-related accumulation of steady-state plasma concentrations of gold consistent with linear pharmacokinetics. In summary, this phase I study was successful in identifying ATM 50 mg intramuscularly weekly as the MTD. Future clinical investigations targeting PKCι are currently in progress.

Impact of disease progression date determination on progression-free survival estimates in advanced lung cancer.

BACKGROUND: In patients with advanced lung cancer, overall survival is largely influenced by progression status. Because progression-free survival (PFS)-based endpoints are controversial, the authors evaluated the impact of the progression date (PD) determination approach on PFS estimates. METHODS: Individual patient data from 21 trials (14 North Central Cancer Treatment Group trials and 7 Southwest Oncology Group trials) were used. The reported PD (RPD) was defined as either the radiographic scan date or the clinical deterioration date. PD was determined using Method 1 (M1), the RPD; M2, 1 day after the last progression-free scan; M3, midpoint between the last progression-free scan and the RPD; and M4, an interval-censoring approach. PFS was estimated using Kaplan-Meier (M1-M3), and maximum-likelihood (M4) methods. Simulation studies were performed to understand the impact of the length of time elapsed between the last progression-free scan and the PD on time-to-progression estimates. RESULTS: PFS estimates using the RPD were the highest, and M2 was the most conservative. M3 and M4 were similar because the majority of progressions occurred during treatment (ie, frequent disease assessments). M3 was influenced less by the length of the assessment schedules (percentage difference from the true time-to-progression, <1.5%) compared with M1 (11% to 30%) and M2 (-8% to -29%). The overall study conclusion was unaffected by the method used for randomized trials. CONCLUSIONS: The magnitude of difference in the PFS estimates was large enough to alter trial conclusions in patients with advanced lung cancer. The results indicate that standards for PD determination, the use of sensitivity analyses, and randomized trials are critical when designing trials and reporting efficacy using PFS-based endpoints.

Phase I study of temsirolimus in combination with EKB-569 in patients with advanced solid tumors.

Purpose Activation of EGFR can stimulate proliferative and survival signaling through mTOR. Preclinical data demonstrates synergistic activity of combined EGFR and mTOR inhibition. We undertook a phase I trial of temsirolimus (T, an mTOR inhibitor) and EKB-569 (E, an EGFR inhibitor) to determine the safety and tolerability. Methods The primary aim was to determine the maximally tolerated dose (MTD) of this combination in adults with solid tumors. Following the dose-escalation phase, (Cohort A), two subsequent cohorts were used to assess any pharmacokinetic (PK) interaction between the agents. Results Forty eight patients were enrolled. The MTD of this combination was E, 35 mg daily and T, 30 mg on days 1-3 and 15-17 using a 28-day cycle. The most common toxicities were nausea, diarrhea, fatigue, anorexia, stomatitis, rash, anemia, neutropenia, thrombocytopenia, and hypertriglyceridemia. Sixteen patients (36%) had at least one grade 3 toxicity. The most frequent grade 3/4 toxicities were diarrhea, dehydration, and nausea and vomiting (19% each). No grade 5 events were seen. Four patients had a partial response and 15 had stable disease. Clinical benefit was seen across a range of tumor types and in all cohorts. PK analysis revealed no significant interaction between E and T. Conclusions This combination of agents is associated with tolerable toxicities at doses that induced responses. PK studies revealed no interaction between the drugs. Further investigations of this targeting strategy may be attractive in renal cell carcinoma, non-small cell lung cancer, alveolar sarcoma, and carcinoid tumor.

Effects of modified atmosphere packaging on the postharvest quality of mulberry leaf vegetable.

Fresh mulberry leaf vegetable is nutritive and becoming popular. However, available preservation technologies are deficient. In present work, the effects of two kinds of modified atmosphere packaging on postharvest quality of fresh mulberry leaf vegetable stored at 4 °C were evaluated. The respiration rate of samples in the modified polyethylene packages (MP20) was 12.88-22.65% lower than that in normal polyethylene packaging (CK). The content of total soluble solids, soluble protein, and total polyphenol in MP20 was less changed than that in CK, and the vitamin C retention was higher as well. Moreover, the lignin content in MP20 was lower than that in CK during storage (19.79% vs 13.38% at day 8), and that was significantly positively related to the polyphenol oxidase and peroxidase activities inhibition. Taken together, a packaging with moderate gas permeability (MP20) is suitable for nutrition maintenance and lignification inhibition of fresh mulberry leaf vegetable during cold storage.

Integration of morphological, physiological and multi-omics analysis reveals a comprehensive mechanism for cuticular wax during development of greasiness in postharvest apples.

Skin greasiness is a common postharvest disorder of apple (Malus × domestica). However, the molecular mechanism of skin greasiness is unclear. In this study, fruits of 'Golden Delicious' (GD), 'Granny Smith', and 'Fuji' with distinct characteristics of greasiness were used for greasiness scoring, wax morphology, wax metabolite, and RNA-seq analyses. Additionally, GD fruit were treated with 1-methylcyclopropene (1-MCP), which repressed greasiness. A partial least squares discriminant analysis (PLS-DA) revealed that wax esters were the critical wax fraction for skin greasiness. Among these wax esters, liquid linoleate esters of short-chain alcohols (C4-C6) and farnesol showed increased contents with increasing greasiness. Their concentrations were significantly correlated with greasiness score. To identify the genes encoding key enzymes for the synthesis of liquid linoleate esters, a weighted gene co-expression network analysis was conducted. MdDCR1, encoding an acyltransferase (defective in cuticular ridges, DCR), was selected as a candidate gene. MdDCR1 was significantly upregulated in greasy skin, and significantly suppressed by 1-MCP treatment. MdDCR1 silencing suppressed the accumulation of liquid linoleate esters of short-chain alcohols, including butyl linoleate, pentyl linoleate, and hexyl linoleate, in GD skin. These results provide insights into the molecular mechanisms of cuticular wax metabolism related to skin greasiness in apple. Our results show that transcriptional regulation of MdDCR1, encoding an acyltransferase that catalyzes the biosynthesis of liquid linoleate esters of short-chain alcohols (C4-C6) via an independent side branch of the C18:2 CoA pathway, regulates the formation of greasiness.

The purported effects of alcohol on appetite and weight in lung cancer patients.

Loss of appetite and weight predict poor outcomes in patients with advanced cancer. Effective and affordable palliative strategies are lacking; but because an emerging non-cancer literature suggests that alcohol can increase appetite and weight, this study explored associations between alcohol and clinical outcomes in lung cancer patients. Among 404 consecutive lung cancer patients enrolled in the Mayo Clinic Lung Cancer Cohort between 2004 and 2008, alcohol consumption (within 6 mo of diagnosis) was as follows: 199 (49%) used none, 158 (14%) were moderate users (7 drinks per wk or less), and 47 (12%) were heavier consumers (more than 7 drinks per wk). Only heavier consumers had a lower likelihood of anorexia (odds ratio: 0.49; 95% CI: 0.25, 0.94; P = 0.03) and weight loss (odds ratio: 0.43; 95% CI: 0.20, 0.91; P = 0.03) compared to those who consumed no alcohol. These conclusions were sustained in multivariate analyses. Neither moderate nor heavier consumption was associated with better or worse survival, although, in univariate analyses, a drop in alcohol consumption was associated with worse survival. This report suggests a need for further study of alcohol as a palliative agent for cancer-associated loss of appetite and weight.

Transcriptional Regulation of Anthocyanin Synthesis by MYB-bHLH-WDR Complexes in Kiwifruit (Actinidia chinensis).

The anthocyanin synthetic pathway is regulated centrally by an MYB-bHLH-WD40 (MBW) complex. Anthocyanin pigmentation is an important fruit quality trait in red-fleshed kiwifruit; however, the underlying regulatory mechanisms involving the MBW complex are not well understood. In this study, one R2R3MYB (AcMYBF110 expressed in fruit characteristically), one bHLH (AcbHLH1), two upstream regulators of AcbHLH1 (AcbHLH4 and AcbHLH5), and one WDR (AcWDR1) are characterized as being involved in the regulation of anthocyanin synthesis in kiwifruit. AcMYBF110 plays an important role in the regulation of anthocyanin accumulation by specifically activating the promoters of several anthocyanin pathway genes including AcCHS, AcF3'H, AcANS, AcUFGT3a, AcUFGT6b, and AcGST1. Coexpression of AcbHLH1, AcbHLH4, or AcbHLH5 together with AcMYBF110 induces much greater anthocyanin accumulation in both tobacco leaves and in Actinidia arguta fruit compared with AcMYBF110 alone. Moreover, this activation is further enhanced by adding AcWDR1. We found that both AcMYBF110 and AcWDR1 interact with all three AcbHLH factors, while AcMYBF110 also interacts with AcWDR1 to form three different MBW complexes that have different regulatory roles in anthocyanin accumulation of kiwifruit. The AcMYBF110-AcbHLH1-AcWDR1 complex directly targets the promoters of anthocyanin synthetic genes. Other features of the regulatory pathways identified include promotion of AcMYBF110, AcbHLH1,and AcWDR1 activities by this MBW complex, providing for both reinforcement and feedback regulation, whereas the AcMYBF110-AcbHLH4/5-AcWDR1 complex is indirectly involved in the regulation of anthocyanin synthesis by activating the promoters of AcbHLH1 and AcWDR1 to amplify the regulation signals of the first MBW complex.

High oxygen atmospheric packaging treatment regulates the postharvest changes of Chinese kale (Brassica oleracea var. alboglabra) during storage.

Chinese kale is one of the most popular vegetables in southern China and Asia, but it has a short shelf-life. The effect of high oxygen atmospheric packaging (HOAP) treatment on the respiration rate as well as chlorophyll content and the expression of their metabolism-related genes of the soluble proteins in Chinese kale during storage were assessed. The results showed that Chinese kale subjected to HOAP treatment showed stimulated respiration rate and regulated expression of chlorophyll metabolism-related genes, such as BrChlases, BrPPH (pheophytin pheophorbide hydrolase), BrPAO (pheidea oxygenase gene), BrRCCR (red chlorophyll catabolite reductase), and BrSAG12 (senescence-associated gene), compared to the Chinese kale in the control. The activities of chlorophyll enzymes, that is, Chlase and Mg-dechelatase, were also influenced by HOAP treatment during storage. Furthermore, the total content of soluble proteins was stimulated to accumulate, and the intensity of protein bands, detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiling, increased in HOAP-treated samples. Based on the current results, as well as the results of our previous study regarding HOAP treatment of other vegetables, we speculate that HOAP may function by regulating the respiration rate and the accumulation of functional proteins, especially chlorophyll catabolic and antioxidant enzymes, to maintain the freshness of Chinese kale during storage. PRACTICAL APPLICATION: HOAP treatment could be a potential method for delaying quality changes and extending the shelf-life of Chinese kale after harvest.

The cancer anorexia/weight loss syndrome: exploring associations with single nucleotide polymorphisms (SNPs) of inflammatory cytokines in patients with non-small cell lung cancer.

OBJECTIVE: The cancer anorexia/weight loss syndrome commonly occurs in patients with non-small cell lung cancer (NSCLC) and is characterized by loss of weight and appetite as well as diminished survival. The current study explored whether any of 22 single nucleotide polymorphisms (SNPs) of certain previously implicated inflammatory cytokines (interleukin-1 beta, interleukin-1RN, interleukin-6, and tumor necrosis factor) are associated with this syndrome. PATIENTS AND METHODS: All NSCLC patients who had been enrolled in the Mayo Clinic Lung Cancer Cohort, had completed a health-related questionnaire approximately 6 months after enrollment, and had blood drawn were included in this study, thus yielding a sample size of 471 patients. RESULTS: Sixty-six (14%) patients manifested weight loss shortly after diagnosis, and 152 (32%) reported appetite loss. Only tumor necrosis factor alpha rs800629 was associated with anorexia (odds ratio: 0.46; 95% confidence interval: 0.29, 0.72; p < 0.001); patients who were heterozygous and minor homozygous were less likely to suffer anorexia. Otherwise, there were no statistically significant associations between any of the other 21 SNPs and weight loss and/or anorexia. In univariate analyses, weight loss, anorexia, more advanced cancer stage, and interleukin-1 beta rs1143627 were associated with a worse survival, and interleukin-6 rs2069835 was associated with better survival. However, in multivariate analyses, cancer stage and patient age were the only statistically significant predictors of worse survival. CONCLUSION: No specific SNP was associated with all aspects of the cancer anorexia/weight loss syndrome, but rs800629 may merit further study in cancer-associated anorexia.

Polyphenol oxidase plays a critical role in melanin formation in the fruit skin of persimmon (Diospyros kaki cv. 'Heishi').

In this study, the melanin in persimmon and its formation were investigated. Melanin was found to be deposited on the cell walls of the upper epidermis and subepidermal cells in persimmon skin and the isolated pigment appears to have lamellar structures. Diagnostic analysis of the isolated pigment showed results that were similar to those of melanin from other sources. Ultraviolet-visible spectroscopy revealed that the extracted skin pigment displayed a broadband, structureless absorption profile that increased progressively towards shorter wavelengths. The Fourier transform infrared spectroscopy assay revealed that melanin in persimmon skin exhibits many characteristic absorption peaks. The phenolic profile analysis suggested that the precursors of this pigment may include gallic acid, procyanidin B1, procyanidin B2, ferulic acid and epigallocatechin gallate. The PPO activity and DkPPO expression significantly increased during melanin formation, and transient overexpression of DkPPO promoted melanin synthesis. These results indicate that the isolated pigment was a type of melanin and that PPO plays a critical role in its formation.

Phenolic compounds and antioxidant activity in red- and in green-fleshed kiwifruits.

Red-fleshed kiwifruits are receiving increasing attention because of their high phenolic contents. However, detailed information on their phenolic compounds and antioxidant capacities remains scarce. Here, six red-fleshed and six green-fleshed kiwifruits were investigated to determine their contents of phenolic compounds and their antioxidant capacities. The results showed chlorogenic acid, p-coumaric acid and ferulic acid were the main phenolic compounds found in kiwifruit. Most of red-fleshed kiwifruits contain higher amounts of total phenolics and anthocyanins, as well as higher activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD). Moreover, they exhibited stronger antioxidant capacities than green-fleshed kiwifruits in ABTS, DPPH and FRAP assays. Furthermore, the reactive oxygen species (ROS) inhibition assay showed the phenolics extracted from red-fleshed kiwifruit can better protect tobacco leaves against hydrogen peroxide (H2O2)-induced oxidative damage. This is because of their abundant anthocyanins which in vitro contribute more to H2O2 scavenging than the other phenolic compounds. Based on these findings, it is fair to conclude the red-fleshed kiwifruits are promising sources of antioxidants in human nutrition.

Natural history of pelvic organ prolapse in postmenopausal women.

OBJECTIVE: To describe the natural history of pelvic organ prolapse and risk factors for changes in vaginal descent in older women. METHODS: This 4-year prospective observational study included 259 postmenopausal women with a uterus enrolled at one Women's Health Initiative clinical site who completed at least two annual pelvic organ prolapse quantification (POP-Q) examinations. We calculated 1-year and 3-year incidence and resolution risks for prolapse (defined as maximal vaginal descent to or beyond the hymen) and estimated progression and regression rates (1 cm or greater and 2 cm or greater changes in maximal vaginal descent) and risk factors. RESULTS: Mean age was 68.1+/-5.5 years, and median vaginal parity was 4. Seventy-three (28%) women had four exams, 128 (49%) had three exams, and 58 (22%) had two exams. Prolapse waxed and waned yearly in individual women. Overall 1-year and 3-year prolapse incidences were 26% (95% confidence interval [CI] 20-33%) and 40% (95% CI 26-56%); 1-year and 3-year prolapse resolution risks were 21% (95% CI 11-33%) and 19% (95% CI 7-39%). Rates of any change in maximal vaginal descent over time varied depending on baseline measurements. Over 3 years, the maximal vaginal descent increased by at least 2 cm in 11.0% (95% CI 4.9-20.5%) of the women and decreased by at least 2 cm in 2.7% (95% CI 0.3-9.5%). Increasing body mass index and grand multiparity increased the risk for vaginal descent progression. CONCLUSION: Prolapse progresses and regresses in older women, although rates of vaginal descent progression are slightly greater than regression overall. Obesity is a risk factor for progression in vaginal descent. LEVEL OF EVIDENCE: III.