Christopher J. Scarlett

Pancreatic cancer remains one of the most lethal of malignancies and a major health burden. We performed whole-genome sequencing and copy number variation (CNV) analysis of 100 pancreatic ductal adenocarcinomas (PDACs). Chromosomal rearrangements leading to gene disruption were prevalent, affecting genes known to be important in pancreatic cancer (TP53, SMAD4, CDKN2A, ARID1A and ROBO2) and new candidate drivers of pancreatic carcinogenesis (KDM6A and PREX2). Patterns of structural variation (variation in chromosomal structure) classified PDACs into 4 subtypes with potential clinical utility: the subtypes were termed stable, locally rearranged, scattered and unstable. A significant proportion harboured focal amplifications, many of which contained druggable oncogenes (ERBB2, MET, FGFR1, CDK6, PIK3R3 and PIK3CA), but at low individual patient prevalence. Genomic instability co-segregated with inactivation of DNA maintenance genes (BRCA1, BRCA2 or PALB2) and a mutational signature of DNA damage repair deficiency. Of 8 patients who received platinum therapy, 4 of 5 individuals with these measures of defective DNA maintenance responded.

Pancreatic cancer is a highly lethal malignancy with few effective therapies. We performed exome sequencing and copy number analysis to define genomic aberrations in a prospectively accrued clinical cohort (n = 142) of early (stage I and II) sporadic pancreatic ductal adenocarcinoma. Detailed analysis of 99 informative tumours identified substantial heterogeneity with 2,016 non-silent mutations and 1,628 copy-number variations. We define 16 significantly mutated genes, reaffirming known mutations (KRAS, TP53, CDKN2A, SMAD4, MLL3, TGFBR2, ARID1A and SF3B1), and uncover novel mutated genes including additional genes involved in chromatin modification (EPC1 and ARID2), DNA damage repair (ATM) and other mechanisms (ZIM2, MAP2K4, NALCN, SLC16A4 and MAGEA6). Integrative analysis with in vitro functional data and animal models provided supportive evidence for potential roles for these genetic aberrations in carcinogenesis. Pathway-based analysis of recurrently mutated genes recapitulated clustering in core signalling pathways in pancreatic ductal adenocarcinoma, and identified new mutated genes in each pathway. We also identified frequent and diverse somatic aberrations in genes described traditionally as embryonic regulators of axon guidance, particularly SLIT/ROBO signalling, which was also evident in murine Sleeping Beauty transposon-mediated somatic mutagenesis models of pancreatic cancer, providing further supportive evidence for the potential involvement of axon guidance genes in pancreatic carcinogenesis.

The diagnosis of pancreatic neuroendocrine tumours (PanNETs) is increasing owing to more sensitive detection methods, and this increase is creating challenges for clinical management. We performed whole-genome sequencing of 102 primary PanNETs and defined the genomic events that characterize their pathogenesis. Here we describe the mutational signatures they harbour, including a deficiency in G:C > T:A base excision repair due to inactivation of MUTYH, which encodes a DNA glycosylase. Clinically sporadic PanNETs contain a larger-than-expected proportion of germline mutations, including previously unreported mutations in the DNA repair genes MUTYH, CHEK2 and BRCA2. Together with mutations in MEN1 and VHL, these mutations occur in 17% of patients. Somatic mutations, including point mutations and gene fusions, were commonly found in genes involved in four main pathways: chromatin remodelling, DNA damage repair, activation of mTOR signalling (including previously undescribed EWSR1 gene fusions), and telomere maintenance. In addition, our gene expression analyses identified a subgroup of tumours associated with hypoxia and HIF signalling. The genomes of 102 primary pancreatic neuroendocrine tumours have been sequenced, revealing mutations in genes with functions such as chromatin remodelling, DNA damage repair, mTOR activation and telomere maintenance, and a greater-than-expected contribution from germ line mutations. Pancreatic neuroendocrine tumours (PanNETs) are the second most common epithelial neoplasm of the pancreas. Aldo Scarpa, Sean Grimmond and colleagues report whole-genome sequencing of 102 primary PanNETs and present analysis of their mutational signatures as part of the International Cancer Genome Consortium. They find frequent mutations in genes with functions that include chromatin remodelling, DNA damage repair, activation of mTOR signalling, and telomere maintenance. They also identify mutational signatures, including one resulting from inactivation of the DNA repair gene MUTYH, and report a larger than expected germline contribution to PanNET development.

Colony-forming units – fibroblast (CFU-Fs), analogous to those giving rise to bone marrow (BM) mesenchymal stem cells (MSCs), are present in many organs, although the relationship between BM and organ-specific CFU-Fs in homeostasis and tissue repair is unknown. Here we describe a population of adult cardiac-resident CFU-Fs (cCFU-Fs) that occupy a perivascular, adventitial niche and show broad trans-germ layer potency in vitro and in vivo. CRE lineage tracing and embryo analysis demonstrated a proepicardial origin for cCFU-Fs. Furthermore, in BM transplantation chimeras, we found no interchange between BM and cCFU-Fs after aging, myocardial infarction, or BM stem cell mobilization. BM and cardiac and aortic CFU-Fs had distinct CRE lineage signatures, indicating that they arise from different progenitor beds during development. These diverse origins for CFU-Fs suggest an underlying basis for differentiation biases seen in different CFU-F populations, and could also influence their capacity for participating in tissue repair.

Starch is the most popular plant polysaccharides, which has been widely used for the development of edible coating films because of its abundance, cost-effectiveness, and excellent film-forming abilities. Starch-based films have good optical, organoleptic and gas barrier properties, however, they have poor mechanical properties. Many attempts have been made to overcome these limitations, such as the addition of co-biopolymers or other secondary additives to improve the mechanical and tensile properties of the films. Properties of the starch-based films can be influenced by many factors, including types of starches, temperature and time during film formation, plasticizers, co-biopolymers, and storage conditions. Understanding the mechanisms of these factors is very important for future studies on the development of starch-based films. This review focuses on starch as a film/coating material and comprehensively discusses the effects of major factors on properties of starch-based films.

Current adjuvant therapies for pancreatic cancer (PC) are inconsistently used and only modestly effective. Because a high proportion of patients who undergo resection for PC likely harbor occult metastatic disease, any adjuvant trials assessing therapies such as radiotherapy directed at locoregional disease are significantly underpowered. Stratification based on the probability (and volume) of residual locoregional disease could play an important role in the design of future clinical trials assessing adjuvant radiotherapy.We assessed the relationships between margin involvement, the proximity to operative resection margins and outcome in a cohort of 365 patients who underwent operative resection for PC.Microscopic involvement of a resection margin by tumor was associated with a poor prognosis. Stratifying the minimum clearance of resection margins by 0.5-mm increments demonstrated that although median survival was no different to clear margins based on these definitions, it was not until the resection margin was clear by more than 1.5 mm that optimal long-term survival was achieved.These data demonstrate that a margin clearance of more than 1.5 mm is important for long-term survival in a subgroup of patients. More aggressive therapeutic approaches that target locoregional disease such as radiotherapy may be beneficial in patients with close surgical margins. Stratification of patients for entry onto future clinical trials based on this criterion may identify those patients who benefit from adjuvant radiotherapy.

BackgroundCurrent staging methods for pancreatic cancer (PC) are inadequate, and biomarkers to aid clinical decision making are lacking. Despite the availability of the serum marker carbohydrate antigen 19.9 (CA19.9) for over two decades, its precise role in the management of PC is yet to be defined, and as a consequence, it is not widely used.MethodsWe assessed the relationship between perioperative serum CA19.9 levels, survival and adjuvant chemotherapeutic responsiveness in a cohort of 260 patients who underwent operative resection for PC.ResultsBy specifically assessing the subgroup of patients with detectable CA19.9, we identified potential utility at key clinical decision points. Low postoperative CA19.9 at 3 months (median survival 25.6 vs 14.8 months, P = 0.0052) and before adjuvant chemotherapy were independent prognostic factors. Patients with postoperative CA 19.9 levels >90 U/ml did not benefit from adjuvant chemotherapy (P = 0.7194) compared with those with a CA19.9 of ≤90 U/ml (median 26.0 vs 16.7 months, P = 0.0108). Normalization of CA19.9 within 6 months of resection was also an independent favorable prognostic factor (median 29.9 vs 14.8 months, P = 0.0004) and normal perioperative CA19.9 levels identified a good prognostic group, which was associated with a 5-year survival of 42%.ConclusionsPerioperative serum CA19.9 measurements are informative in patients with detectable CA19.9 (defined by serum levels of >5 U/ml) and have potential clinical utility in predicting outcome and response to adjuvant chemotherapy. Future clinical trials should prioritize incorporation of CA19.9 measurement at key decision points to prospectively validate these findings and facilitate implementation.

The leaves of Carica papaya are generally considered waste but their extracts have been linked with various health benefits. This study aimed to optimize extraction conditions and determine the effect of aqueous extraction on the yield of polyphenols from papaya leaves. The efficiency of water extraction was compared to the organic solvents acetone, ethanol and methanol. A method to prepare crude powder from the leaves was developed, with its composition and antioxidant properties also examined. We show that temperature, extraction time and water-to-leaf ratio had significant effects on the extracted polyphenol yield as well as the scavenging and total antioxidant activities. Optimal extraction conditions were 70 °C for 20 min, with a water-to-leaf ratio of 100:7.5 mL/g. Higher levels of polyphenols were extracted using water in comparison to the organic solvents, while ethanol extraction provided the highest level of saponins. A simple and scalable method was developed to obtain approximately 190 g of powder from 1 kg of dried papaya leaves. The crude powder contained 6.3% polyphenols, and when compared to butylated hydroxytoluene (BHT), Vitamins C and E and epigallocatechin gallate (EGCG) had lower scavenging and total antioxidant activity. However, as this method used water for extraction, it is considered safe and offers great potential for further purification and application in future studies.

A large quantity of bananas is produced annually and its peel, which accounts for about one third of the fruit weight, is mostly discarded as waste. The peel has been traditionally used for the treatment of various ailments. This by-product is rich in phenolics with over 40 individual compounds identified. However, composition and levels of these compounds are influenced by various factors, including varieties, maturity, cultivation conditions, and pre-treatments. Phenolics within banana peels have been found to possess potent antioxidant and antimicrobial properties, and linked with various health benefits. Therefore, it is worthwhile to recover phenolics from this by-product for further utilisation in food and pharmaceutical industries. This review comprehensively highlights the phenolic compounds as well as major factors affecting their presence within the banana peel, reviews the current applications of this by-product, outlines its potential uses in food and pharmaceutical industries and finally proposes a trend for future studies.

Pancreatic cancer is molecularly diverse, with few effective therapies. Increased mutation burden and defective DNA repair are associated with response to immune checkpoint inhibitors in several other cancer types. We interrogated 385 pancreatic cancer genomes to define hypermutation and its causes. Mutational signatures inferring defects in DNA repair were enriched in those with the highest mutation burdens. Mismatch repair deficiency was identified in 1% of tumors harboring different mechanisms of somatic inactivation of MLH1 and MSH2. Defining mutation load in individual pancreatic cancers and the optimal assay for patient selection may inform clinical trial design for immunotherapy in pancreatic cancer. Pancreatic cancer is molecularly diverse, with few effective therapies. Increased mutation burden and defective DNA repair are associated with response to immune checkpoint inhibitors in several other cancer types. We interrogated 385 pancreatic cancer genomes to define hypermutation and its causes. Mutational signatures inferring defects in DNA repair were enriched in those with the highest mutation burdens. Mismatch repair deficiency was identified in 1% of tumors harboring different mechanisms of somatic inactivation of MLH1 and MSH2. Defining mutation load in individual pancreatic cancers and the optimal assay for patient selection may inform clinical trial design for immunotherapy in pancreatic cancer. Pancreatic ductal adenocarcinoma has a 5-year survival of <5%, with therapies offering only incremental benefit,1Vogelzang N.J. et al.J Clin Oncol. 2012; 30: 88-109Crossref PubMed Scopus (85) Google Scholar potentially due to the diversity of its genomic landscape.2Bailey P. et al.Nature. 2016; 531: 47-52Crossref PubMed Scopus (1973) Google Scholar, 3Biankin A.V. et al.Nature. 2012; 491: 399-405Crossref PubMed Scopus (1379) Google Scholar, 4Waddell N. et al.Nature. 2015; 518: 495-501Crossref PubMed Scopus (1466) Google Scholar Recent reports link high mutation burden with response to immune checkpoint inhibitors in several cancer types.5Le D.T. et al.N Engl J Med. 2015; 372: 2509-2520Crossref PubMed Scopus (6099) Google Scholar Defining tumors that are hypermutated with an increased mutation burden and understanding the underlying mechanisms in pancreatic cancer has the potential to advance therapeutic development, particularly for immunotherapeutic strategies. Whole genome sequencing (WGS, n = 180) and whole exome sequencing (n = 205) of 385 unselected predominantly sporadic pancreatic ductal adenocarcinoma (Supplementary Table 1) defined a mean mutation load of 1.8 and 1.1 mutation per megabase (Mb), respectively (Supplementary Table 2). Outlier analysis identified 20 tumors with the highest mutation burden (5.2%, 15 WGS and 5 exome) (Table 1 and Supplementary Figure 1A), 5 of which were considered extreme outliers and classified as hypermutated as they contained ≥12 somatic mutations/Mb, the defined threshold for hypermutation in colorectal cancer.6Cancer Genome Atlas NetworkNature. 2012; 487: 330-337Crossref PubMed Scopus (5894) Google Scholar Immunohistochemistry for mismatch repair (MMR) proteins (MSH2, MSH6, MLH1, and PMS2) identified 4 MMR-deficient tumors, all of which were hypermutated (n = 180, Figure 1).Table 1Clinical and Histologic Features and Proposed Etiology for Highly Mutated Pancreatic Ductal Adenocarcinoma Tumors (n = 20)Sample IDPersonal and family history of malignancyHistologyMutation load, mutations/MbIHC resultMSIsensor scoreKRAS mutationPredominant mutation signature (mutations/Mb)SV subtype (no. of events)Proposed etiologyHypermutation (extreme outliers) ICGC_0076aSample sequenced by WGS, other samples by exome sequencing.NoneMixed signet ring, mucinous and papillary adenocarcinoma38.55Absent MLH1 and PMS228.3p.G12VMMR (18.3)Scattered (131)MMR deficiency: >280 kb somatic homozygous deletion over MSH2. ICGC_0297aSample sequenced by WGS, other samples by exome sequencing.NoneUndifferentiated adenocarcinoma60.62Absent MSH2 and MSH627.33WTMMR (33.4)Scattered (75)MMR deficiency: Somatic MLH1 promoter hypermethylation. ICGC_0548aSample sequenced by WGS, other samples by exome sequencing.NoneDuctal adenocarcinoma, moderately differentiated30.13Absent MSH2 and MSH617.47WTMMR (16.6)Stable (49)MMR deficiency: >27 kb somatic inversion rearrangement disrupting MSH2. ICGC_0328aSample sequenced by WGS, other samples by exome sequencing.NoneDuctal adenocarcinoma16.63Normal3.2p.G12DUnknown (11.9)Scattered (110)Cell line with signature: etiology unknown. ICGC_00901 FDR, father CRCDuctal adenocarcinoma, moderately differentiated12.9Absent MSH2 and MSH60.21p.G12CNANAMMR deficiency: somatic MSH2 splice site c.2006G>A.Highly mutated tumors ICGC_0054aSample sequenced by WGS, other samples by exome sequencing.NoneDuctal adenocarcinoma, poorly differentiated6.52Normal0.01p.G12VHR deficiency (1.3)Unstable (310)HR deficiency: no germline or somatic cause found. ICGC_0290aSample sequenced by WGS, other samples by exome sequencing.NoneDuctal adenocarcinoma, poorly differentiated6.54Not available0.07p.G12VHR deficiency (3.1)Unstable (558)HR deficiency: Germline BRCA2 mutation c.7180A>T, p.A2394*. Somatic CN-LOH. ICGC_0215aSample sequenced by WGS, other samples by exome sequencing.2 FDR lung cancer, 2 FDR prostate cancer. Previous CRC and melanomaDuctal adenocarcinoma, moderately differentiated6.27Normal0.01p.G12VHR deficiency (1.9)Scattered (111)HR deficiency: Germline ATM mutation c.7539_7540delAT, p.Y2514*. Somatic CN-LOH. ICGC_0324NoneDuctal adenocarcinoma, moderately differentiated6.24Normal0p.G12DNANAUndefined ICGC_0034aSample sequenced by WGS, other samples by exome sequencing.NoneDuctal adenocarcinoma, poorly differentiated6.09Normal4.02p.G12DHR deficiency (3.4)Unstable (366)HR deficiency: Germline BRCA2 mutation c.5237_5238insT, p.N1747*. Somatic CN-LOH. ICGC_0131aSample sequenced by WGS, other samples by exome sequencing.Lung cancer after PCDuctal adenocarcinoma, moderately differentiated5.63Normal0p.G12DT>G at TT sites (3.0)Focal (147)T>G at TT sites signature: etiology potentially associated with DNA oxidation ICGC_0006aSample sequenced by WGS, other samples by exome sequencing.1 FDR, father lung cancerAdenocarcinoma arising from IPMN, moderately differentiated5.29Normal0.01p.G12DHR deficiency (1.2)Unstable (211)HR deficiency: Somatic BRCA2 c.5351dupA, p.N1784KfsTer3. Somatic CN-LOH. ICGC_0321aSample sequenced by WGS, other samples by exome sequencing.2 FDR, mother and cousin breast cancerDuctal adenocarcinoma, poorly differentiated4.79Not available0p.G12DHR deficiency (2.1)Unstable (286)HR deficiency: Germline BRCA2 c.6699delT, p.F2234LfsTer7. Somatic CN loss- 1 copy. ICGC_0309aSample sequenced by WGS, other samples by exome sequencing.NoneAdenocarcinoma arising from IPMN, moderately differentiated4.74Normal0.03p.G12VT>G at TT sites (3.1)Unstable (232)T>G at TT sites signature: etiology potentially associated with DNA oxidation ICGC_0005aSample sequenced by WGS, other samples by exome sequencing.1 FDR, mother CRCDuctal adenocarcinoma, poorly differentiated4.72Not available1p.G12VHR deficiency (1.1)Focal (95)HR deficiency: No germline or somatic cause found. ICGC_0016aSample sequenced by WGS, other samples by exome sequencing.NoneDuctal adenocarcinoma, poorly differentiated4.61Normal3.03p.G12VHR deficiency (1.7)Unstable (447)HR deficiency: potentially linked to Somatic RPA1 c.273G>T, p.R91S ICGC_00461 FDR, brother PCDuctal adenocarcinoma, poorly differentiated4.3Normal0p.Q61HNANAUndefined GARV_0668aSample sequenced by WGS, other samples by exome sequencing.NoneDuctal adenocarcinoma, poorly differentiated4.3Not available2.19p.G12VHR deficiency (1.6)Unstable (464)HR deficiency: Germline BRCA2 c.7068_7069delTC, p.L2357VfsTer2. Somatic CN loss - 1 copy. ICGC_0291NoneDuctal adenocarcinoma, well differentiated3.84Not available0.03p.G12RNANAHR deficiency: Somatic BRCA2 c.7283T>A, p.L2428*. ICGC_0256NoneDuctal adenocarcinoma, poorly differentiated3.72Not available0.06p.G12DNANAUndefinedCRC, colorectal cancer; FDR, first-degree relative; IHC, immunohistochemistry; IPMN, intraductal papillary mucinous neoplasm; CN-LOH, copy neutral loss of heterozygosity; CN, copy number; PC, pancreatic cancer; NA, not applicable to exome data.a Sample sequenced by WGS, other samples by exome sequencing. Open table in a new tab CRC, colorectal cancer; FDR, first-degree relative; IHC, immunohistochemistry; IPMN, intraductal papillary mucinous neoplasm; CN-LOH, copy neutral loss of heterozygosity; CN, copy number; PC, pancreatic cancer; NA, not applicable to exome data. KRAS mutation status and histopathologic characteristics have been associated with MMR-deficient pancreatic tumors.7Goggins M. et al.Am J Pathol. 1998; 152: 1501-1507PubMed Google Scholar Of the 4 MMR-deficient tumors in our cohort, 2 were KRAS wild-type; 3 had undifferentiated to moderately differentiated histology and one had a signet-ring component. These features were not predictive of MMR deficiency in our cohort, as 11 additional non−MMR-deficient tumors had a signet-ring cell component or colloid morphology, and 131 of 347 assessable tumors had poorly or undifferentiated histology. Mutational signature analysis can detect MMR deficiency indirectly based on the pattern of somatic mutations.8Alexandrov L.B. et al.Nature. 2013; 500: 415-421Crossref PubMed Scopus (6213) Google Scholar An MMR-deficient signature dominated the MMR-deficient tumors (with WGS), and was minimal in MMR intact tumors (Supplementary Figure 1). In addition, microsatellite instability (MSI), a hallmark of MMR deficiency in colorectal cancer, was detected in all three MMR deficient tumors with WGS using MSIsensor9Niu B. Ye K. et al.Bioinformatics. 2014; 30: 1015-1016Crossref PubMed Scopus (294) Google Scholar (Supplementary Table 2). MSI was not identified for the fourth MMR deficient sample potentially due to the reduced number of microsatellite loci in exome data. The underlying causes of MMR deficiency in the 4 cases were private somatic events. For 2 cases, MSH2 was disrupted by different structural rearrangements, 1 case contained a missense MSH2 mutation and the last, methylation of the MLH1 promoter (Figure 1). The missense mutation caused an MSH2 splice acceptor site mutation that alters the same nucleotide results in a pathogenic skipping of exon 13 in germline studies.10Thompson B.A. et al.Nat Genet. 2014; 46: 107-115Crossref PubMed Scopus (346) Google Scholar Hypermethylation of the MLH1 promoter is the predominant mechanism of MSI in sporadic colon cancer.11Boland C.R. et al.Gastroenterology. 2010; 138: 2073-2087 e3Abstract Full Text Full Text PDF PubMed Scopus (1359) Google Scholar The remaining hypermutated tumor contained an intact MMR pathway, and was a cell line (ATCC, CRL-2551) with an unidentified mutational signature, therefore the high mutation burden in this sample may be the result of long-term cell culture. The 15 samples (11 WGS and 4 exome) identified in the outlier analysis with high mutation burden, but not hypermutated (∼4 to 12 mutations/Mb) contained no evidence of MMR deficiency. Mutational signature analysis of the WGS samples indicated homologous recombination (HR) repair deficiency as the most substantial (range, 1.0–3.4 mutations/Mb) contributor to the mutation burden for 8 WGS mutation load outlier tumors. In support of a HR defect4Waddell N. et al.Nature. 2015; 518: 495-501Crossref PubMed Scopus (1466) Google Scholar; 7 of these tumors contained high levels of genomic instability with >200 structural variants and mutations in genes involved in HR were present for 6 of 8 cases (Supplementary Table 2). In addition, 1 case that had undergone exome sequencing had a somatic BRCA2 nonsense mutation that likely contributed to HR deficiency in this case. A mutational signature associated with T>G mutations at TT sites previously described in other cancers, including esophageal cancer12Nones K. Waddell N. Wayte N. et al.Nat Commun. 2014; : 5Google Scholar was the major contributor (>3 mutations/Mb) in 2 samples. For these 2 and the remaining 4 cases, no potential causative event could be identified. Although germline defects in MMR genes are well reported in pancreatic cancer13Grant R.C. Selander I. et al.Gastroenterology. 2015; 148: 556-564Abstract Full Text Full Text PDF PubMed Scopus (211) Google Scholar in our cohort, they did not contribute to MMR deficiency even in those with familial pancreatic cancer or a personal or family history of Lynch-related tumors. A germline truncating variant was detected in PMS2 in 1 case, but did not have loss of the second allele, had normal immunohistochemistry staining and did not display a MMR mutational signature (Supplementary Table 2). MMR deficiency is important in the evolution in a small, but meaningful proportion of pancreatic cancers with a prevalence of 1% (4 of 385) in our cohort. This is consistent with recent studies using the Bethesda polymerase chain reaction panel,14Laghi L. et al.PLoS One. 2012; 7: e46002Crossref PubMed Scopus (55) Google Scholar and with previous estimates of MSI prevalence of 2%−3%.15Nakata B. et al.Clin Cancer Res. 2002; 8: 2536-2540PubMed Google Scholar However, in tumors with low epithelial content that underwent exome sequencing, the sensitivity of somatic mutation detection is reduced, which will affect mutation burden and signature analysis. While cognizant of small numbers, immunohistochemistry was the most accurate in defining MMR due to multiple genomic mechanisms of MMR gene inactivation. Multiple methods to define MMR deficiency may be required for clinical trials that aim to recruit MMR-deficient participants to assess the potential efficacy of checkpoint inhibitors or other therapies in pancreatic cancer. Homologous recombination-deficient tumors, and those with a novel signature seen in esophageal cancer had an increased mutation burden, and need further evaluation as potential patient selection markers for clinical trials of checkpoint inhibitor and other therapies that target tumors with a high mutation burden. The authors would like to thank Cathy Axford, Deborah Gwynne, Mary-Anne Brancato, Clare Watson, Michelle Thomas, Gerard Hammond, and Doug Stetner for central coordination of the Australian Pancreatic Cancer Genome Initiative, data management, and quality control; Mona Martyn-Smith, Lisa Braatvedt, Henry Tang, Virginia Papangelis, and Maria Beilin for biospecimen acquisition; and Sonia Grimaldi and Giada Bonizzato of the ARC-Net Biobank for biospecimen acquisition. For a full list of contributors see Australian Pancreatic Cancer Genome Initiative: http://www.pancreaticcancer.net.au/apgi/collaborators. The cohort consisted of 385 patients with histologically verified pancreatic exocrine carcinoma, prospectively recruited between 2006 and 2013 through the Australian Pancreatic Cancer Genome Initiative (www.pancreaticcancer.net.au) as part of the International Cancer Genome Consortium.1Hudson T.J. et al.Nature. 2010; 464: 993-998Crossref PubMed Scopus (1689) Google Scholar Ethical approval was granted at all treating institutions and individual patients provided informed consent upon entry to the study. The clinicopathologic information for the cohort is described in (Supplementary Table 1), and the global mutation profile has previously been reported for some of these tumors (Supplementary Table 2). Tumor and normal DNA were extracted after histologic review from fresh frozen tissue samples collected at the time of surgical resection or biopsy, as described previously.2Biankin A.V. et al.Nature. 2012; 491: 399-405Crossref PubMed Scopus (1513) Google Scholar Tumor cellularity was determined from single-nucleotide polymorphism array data using qpure.3Song S. et al.PLoS One. 2012; 7: e45835Crossref PubMed Scopus (85) Google Scholar Tumors with epithelial content ≥40% underwent WGS lower cellularity tumors underwent whole exome sequencing. DNA from patient-derived pancreas cell lines and matched normal was also extracted. Exome and WGS were performed using paired 100-bp reads on the Illumina HiSeq 2000, as described previously.2Biankin A.V. et al.Nature. 2012; 491: 399-405Crossref PubMed Scopus (1513) Google Scholar, 4Waddell N. et al.Nature. 2015; 518: 495-501Crossref PubMed Scopus (1686) Google Scholar Regions of germline and somatic copy number change were detected using Illumina SNP BeadChips with GAP.5Popova T. et al.Genome Biol. 2009; 10 (R128−R128)Crossref PubMed Scopus (151) Google Scholar Somatic structural variants were identified from WGS reads using the qSV tool.4Waddell N. et al.Nature. 2015; 518: 495-501Crossref PubMed Scopus (1686) Google Scholar, 6Patch A.M. et al.Nature. 2015; 521: 489-494Crossref PubMed Scopus (930) Google Scholar Single nucleotide variants were called using 2 variant callers: qSNP7Kassahn K.S. et al.PLoS One. 2013; 8: e74380Crossref PubMed Scopus (52) Google Scholar and GATK.8McKenna A. et al.Genome Res. 2010; 20: 1297-1303Crossref PubMed Scopus (14755) Google Scholar Mutations identified by both callers or, those that were unique to a caller but verified by an orthogonal sequencing approach, were considered high confidence and used in all subsequent analyses. Small indels (<200 bp) were identified using Pindel9Ye K. et al.Bioinformatics. 2009; 25: 2865-2871Crossref PubMed Scopus (1391) Google Scholar and each indel was visually inspected in the Integrative Genome Browser. The distribution of the total number of small somatic mutations (coding and noncoding single nucleotide and indel variants) identified per megabase for exome and WGS sequence data were analyzed separately. The group of samples with high mutation load, at the top of each distribution, were defined as the upper distribution outliers for mutations per megabase, that is, ≥75th centile + (1.5× interquartile range). The threshold for detecting outliers in the exome and WGS groups was 3.4 and 4.2 mutations/Mb, respectively. From within the highly mutated set of tumors, hypermutated samples were identified as those with a mutation rate exceeding the thresholds for extreme distribution outliers (≥75th centile + [5× interquartile range]) of 7.4 and 8.1 mutations/Mb for exome and WGS sequencing, respectively. MSIsensor was used to detect microsatellite instability by directly comparing microsatellite repeat lengths between paired normal and tumor sequencing data.10Niu B. et al.Bioinformatics. 2014; 30: 1015-1016Crossref PubMed Scopus (378) Google Scholar A MSIsensor score of >3.5% of somatic microsatellites with repeat length shifts was the detection threshold used to indicate microsatellite instability as published for endometrial cancer.10Niu B. et al.Bioinformatics. 2014; 30: 1015-1016Crossref PubMed Scopus (378) Google Scholar This correlated well with the 5 and 7 microsatellite panels recommended in the Bethesda guidelines.10Niu B. et al.Bioinformatics. 2014; 30: 1015-1016Crossref PubMed Scopus (378) Google Scholar, 11Umar A. et al.J Natl Cancer Inst. 2004; 96: 261-268Crossref PubMed Scopus (2461) Google Scholar Tissue microarrays were constructed using at least three 1-mm formalin-fixed, paraffin-embedded tumor cores. Immunohistochemistry for MSH6 and PMS2 proteins was performed on tissue microarray sections as a screen for MMR deficiency due to MMR proteins forming heterodimers with concordant mismatch repair loss (ie, loss of MLH1 and PMS2 or loss of MSH2 and MSH6).12Hall G. et al.Pathology. 2010; 42: 409-413Abstract Full Text PDF PubMed Scopus (98) Google Scholar Immunohistochemistry on full tumor sections for MSH2, MLH1, MSH6, and PMS2 was performed in those with abnormal staining in core sections. The immunohistochemistry was performed as described previously12Hall G. et al.Pathology. 2010; 42: 409-413Abstract Full Text PDF PubMed Scopus (98) Google Scholar and scored by a senior pathologist. Somatic mutational signatures were extracted from the whole genome sequenced samples using the framework described previously.13Alexandrov L.B. et al.Cell Rep. 2013; 3: 246-259Abstract Full Text Full Text PDF PubMed Scopus (734) Google Scholar High confidence somatic substitutions were classified by the substitution change and sequence context, that is, the type of immediately neighboring bases to the variant. The framework processes the counts of somatic mutations at each context within each sample using non-negative factorization to produce the different signature profiles that are present in the data. The profiles identified were matched against reported signatures from the Cancer of Somatic Mutations in Cancer (http://cancer.sanger.ac.uk/cosmic/signatures). The major contributory signatures, defined as the mutational signature with the highest number of contributing somatic substitution variants, is reported for highly mutated whole genome samples. Bisulfite-converted whole-genome amplified DNA was hybridized to Infinium Human Methylation 450K Beadchips according to the manufacturers protocol (Illumina). Methylation arrays were performed on DNA from 174 pancreatic ductal adenocarcinoma samples, which were compared to DNA from 29 adjacent nonmalignant pancreata. A subset of the methylation data has been published previously.14Nones K. et al.Int J Cancer. 2014; 135: 1110-1118Crossref PubMed Scopus (156) Google Scholar We examined the data for evidence of tumor-specific hypermethylation of the promoter region of MLH1 and MSH2 genes. The methylation array data have been deposited into the International Cancer Genome Consortium data portal (dcc.icgc.org, project PACA-AU). Download .xlsx (.08 MB) Help with xlsx files Supplementary Tables 1 and 2

This study aimed to study the impact of selected common organic solvents on extractable solids, phytochemical composition, and antioxidant capacity of S. chinensis . The results showed that the tested solvents played an important role in extraction of total solid and phytochemical composition as well as antioxidant capacity of S. chinensis . Acetone (50% v/v) was found to be the optimal extraction solvent for extractable solids (12.2%), phenolic compounds (60 mg GAE/g DW), flavonoids (100 mg CE/g DW), proanthocyanidins (47.4 mg CE/g DW), and saponins (754 mg EE/g DW) as well as antioxidant capacity (ABTS 334 mM TE/g DW, DPPH 470 mM TE/g DW, FRAP 347 mM TE/g DW, and CUPRAC 310 mM TE/g DW). The extract prepared from 50% acetone had high levels of bioactive compounds (TPC 555 mg GAE/g CRE, flavonoids 819 mg CE/g CRE, proanthocyanidins 392 mg CE/g CRE, and saponins 1,880 mg EE/g CRE) as well as antioxidant capacity (ABTS 414 mM TE/g, DPPH 407 mM TE/g, FRAP 320 mg TE/g, and CUPRAC 623 mM TE/g), thus further confirming that 50% acetone is the solvent of choice. Therefore, 50% acetone is recommended for extraction of phenolic compounds, their secondary metabolites, saponins, and antioxidant capacity from the root of S. chinensis for further isolation and utilisation.

The influence of process variables (pea starch, guar gum and glycerol) on the viscosity (V), solubility (SOL), moisture content (MC), transparency (TR), Hunter parameters (L, a, and b), total color difference (ΔE), yellowness index (YI), and whiteness index (WI) of the pea starch based edible films was studied using three factors with three level Box–Behnken response surface design. The individual linear effect of pea starch, guar and glycerol was significant (p < 0.05) on all the responses. However, a value was only significantly (p < 0.05) affected by pea starch and guar gum in a positive and negative linear term, respectively. The effect of interaction of starch × glycerol was also significant (p < 0.05) on TR of edible films. Interaction between independent variables starch × guar gum had a significant impact on the b and YI values. The quadratic regression coefficient of pea starch showed a significant effect (p < 0.05) on V, MC, L, b, ΔE, YI, and WI; glycerol level on ΔE and WI; and guar gum on ΔE and SOL value. The results were analyzed by Pareto analysis of variance (ANOVA) and the second order polynomial models were developed from the experimental design with reliable and satisfactory fit with the corresponding experimental data and high coefficient of determination (R2) values (>0.93). Three-dimensional response surface plots were established to investigate the relationship between process variables and the responses. The optimized conditions with the goal of maximizing TR and minimizing SOL, YI and MC were 2.5 g pea starch, 25% glycerol and 0.3 g guar gum. Results revealed that pea starch/guar gum edible films with appropriate physical and optical characteristics can be effectively produced and successfully applied in the food packaging industry.

Novel edible composite coatings based on pea starch and guar gum (PSGG), PSGG blended with lipid mixture containing the hydrophobic compounds shellac and oleic acid (PSGG-Sh), and a layer-by-layer (LBL) approach (PSGG as an internal layer and shellac as an external layer), were investigated and compared with a commercial wax (CW) and uncoated fruit on postharvest quality of ‘Valencia’ oranges held for up to four weeks at 20 °C and 5 °C with an additional storage for 7 d at 20 °C. The incorporation of lipid compounds into the PSGG coatings (PSGG-Sh) generally resulted in the best performance in reducing fruit respiration rate, ethylene production, weight and firmness loss, peel pitting, and fruit decay rate of the coated oranges. Fruit coated with PSGG-Sh and a single layer PSGG coatings generally resulted in higher scores for overall flavor and freshness after four weeks at 5 °C followed by one week at 20 °C than uncoated fruit, as assessed by a sensory panel. Although the LBL coating reduced weight loss and respiration rate with improved firmness retention to a greater extent than the single layer PSGG coating, the bilayer coating also resulted in higher levels of ethanol causing increased perception of off-flavors. Overall results suggested that PSGG-based edible coatings could be a beneficial substitute to common commercial waxes for maintaining quality and storability, as well as extending shelf life of citrus fruit and potentially other fresh horticultural produce.

A rice starch edible coating blended with sucrose esters was developed for controlling the postharvest physiological activity of Cavendish banana to extend postharvest quality during ripening at 20 ± 2 °C. Coating effectiveness was assessed against changes in fruit physiochemical parameters such as weight loss, titratable acidity, total soluble solids, flesh fruit firmness, ion leakage, colour change, respiration, ethylene production, chlorophyll degradation and starch conversion were determined. The topography of coating material on the fruit surface was evaluated by scanning electron microscope (SEM). Surface morphology studies highlighted the binding compatibility of the coating matrix with the fruit peel character and formed a continuous uniform layer over the fruit surface. The results showed that the coating was effective in delaying ethylene biosynthesis and reducing respiration rate. Other factors impacting included delayed chlorophyll degradation, reduced weight loss and retention of fruit firmness for the first six days, all of which improved the commercial value of the fruit. The shelf life of coated fruit was prolonged for 12 days in comparison with the untreated control which ripened within seven days and lost marketability after Day 6. The pilot study demonstrates the effectiveness of a starch-based edible coating formulation for improving the ambient storage capacity of banana fruit.

Myc oncoproteins are commonly upregulated in human cancers of different organ origins, stabilized by Aurora A, degraded through ubiquitin-proteasome pathway-mediated proteolysis, and exert oncogenic effects by modulating gene and protein expression. Histone deacetylases are emerging as targets for cancer therapy. Here we demonstrated that the class III histone deacetylase SIRT2 was upregulated by N-Myc in neuroblastoma cells and by c-Myc in pancreatic cancer cells, and that SIRT2 enhanced N-Myc and c-Myc protein stability and promoted cancer cell proliferation. Affymetrix gene array studies revealed that the gene most significantly repressed by SIRT2 was the ubiquitin-protein ligase NEDD4. Consistent with this finding, SIRT2 repressed NEDD4 gene expression by directly binding to the NEDD4 gene core promoter and deacetylating histone H4 lysine 16. Importantly, NEDD4 directly bound to Myc oncoproteins and targeted Myc oncoproteins for ubiquitination and degradation, and small-molecule SIRT2 inhibitors reactivated NEDD4 gene expression, reduced N-Myc and c-Myc protein expression, and suppressed neuroblastoma and pancreatic cancer cell proliferation. Additionally, SIRT2 upregulated and small-molecule SIRT2 inhibitors decreased Aurora A expression. Our data reveal a novel pathway critical for Myc oncoprotein stability, and provide important evidences for potential application of SIRT2 inhibitors for the prevention and therapy of Myc-induced malignancies.

Purpose Individuals with adenocarcinoma of the ampulla of Vater demonstrate a broad range of outcomes, presumably because these cancers may arise from any one of the three epithelia that converge at that location. This variability poses challenges for clinical decision making and the development of novel therapeutic strategies. Patients and Methods We assessed the potential clinical utility of histomolecular phenotypes defined using a combination of histopathology and protein expression (CDX2 and MUC1) in 208 patients from three independent cohorts who underwent surgical resection for adenocarcinoma of the ampulla of Vater. Results Histologic subtype and CDX2 and MUC1 expression were significant prognostic variables. Patients with a histomolecular pancreaticobiliary phenotype (CDX2 negative, MUC1 positive) segregated into a poor prognostic group in the training (hazard ratio [HR], 3.34; 95% CI, 1.69 to 6.62; P &lt; .001) and both validation cohorts (HR, 5.65; 95% CI, 2.77 to 11.5; P &lt; .001 and HR, 2.78; 95% CI, 1.25 to 7.17; P = .0119) compared with histomolecular nonpancreaticobiliary carcinomas. Further stratification by lymph node (LN) status defined three clinically relevant subgroups: one, patients with histomolecular nonpancreaticobiliary (intestinal) carcinoma without LN metastases who had an excellent prognosis; two, those with histomolecular pancreaticobiliary carcinoma with LN metastases who had a poor outcome; and three, the remainder of patients (nonpancreaticobiliary, LN positive or pancreaticobiliary, LN negative) who had an intermediate outcome. Conclusion Histopathologic and molecular criteria combine to define clinically relevant histomolecular phenotypes of adenocarcinoma of the ampulla of Vater and potentially represent distinct diseases with significant implications for current therapeutic strategies, the ability to interpret past clinical trials, and future trial design.

The N-Myc oncoprotein is a critical factor in neuroblastoma tumorigenesis which requires additional mechanisms converting a low-level to a high-level N-Myc expression. N-Myc protein is stabilized when phosphorylated at Serine 62 by phosphorylated ERK protein. Here we describe a novel positive feedback loop whereby N-Myc directly induced the transcription of the class III histone deacetylase SIRT1, which in turn increased N-Myc protein stability. SIRT1 binds to Myc Box I domain of N-Myc protein to form a novel transcriptional repressor complex at gene promoter of mitogen-activated protein kinase phosphatase 3 (MKP3), leading to transcriptional repression of MKP3, ERK protein phosphorylation, N-Myc protein phosphorylation at Serine 62, and N-Myc protein stabilization. Importantly, SIRT1 was up-regulated, MKP3 down-regulated, in pre-cancerous cells, and preventative treatment with the SIRT1 inhibitor Cambinol reduced tumorigenesis in TH-MYCN transgenic mice. Our data demonstrate the important roles of SIRT1 in N-Myc oncogenesis and SIRT1 inhibitors in the prevention and therapy of N-Myc-induced neuroblastoma.

The influence of different plasticizers (glycols, sugars and polyols) on the moisture sorption, mechanical, physical, optical, and microstructure characteristics of pea starch-guar gum (PSGG) film was studied. All plasticizers formed homogeneous, transparent, and smooth films, while PEG-400 did not produce film with suitable characteristics. Fourier transform infrared (FTIR) spectroscopy results indicated some interaction between plasticizers and the polymers. Scanning electron microscopy (SEM) observations of the films presented surfaces without cracks, breaks, or openings which were indicator of the miscibility and compatibility of employed plasticizers with PSGG films. The results showed that the films containing plasticizers with higher functional groups had lower equilibrium moisture content at aw <0.4. In general, a reduction in tensile strength and Young's modulus and an increase in elongation at break were detected when molecular weight of plasticizers and relative humidity increased in all film formulations. Films plasticized with monosaccharide showed similar mechanical properties to those with sorbitol, but lower solubility and water vapour permeability (WVP), higher transparency and moisture content than the sorbitol-plasticized films. The most noticeable plasticization effect was exerted by following order: glycerol > EG > PG > xylitol > fructose > sorbitol > mannitol > galactose > glucose > sucrose > maltitol.

Eucalyptus robusta (E. robusta) has a significant value in traditional medicine and recently has been shown to possess many pharmacological properties in vitro. This study was designed to utilise microwave-assisted extraction (MAE) to yield optimal total phenolic content (TPC), total flavonoid content (TFC), proanthocyanidin levels and antioxidant capacity from E. robusta using water as the solvent, facilitated by the use of response surface methodology (RSM). A three-level-three-factor Box–Behnken design was implemented to elucidate the effect of irradiation time, power and sample-to-solvent ratio on the yields of these phytochemicals. The results highlighted the accuracy and reliability of RSM as a tool for predicting the yields of TPC, TFC, proanthocyanidins and total antioxidants using MAE. Sample-to-solvent ratio had the greatest impact on the TPC yield followed by power and irradiation time. The optimal MAE conditions for TPC and TFC were 3 min, 600 W power and 2 g/100 mL sample-to-solvent ratio. The experimental yield of TPC was 58.40 ± 1.03 mg GAE/g, and 19.15 ± 1.06 mg RE/g of TFC was obtained under these optimal conditions. These conditions, optimised for maximum TPC yield also liberated 62%, 64.6%, 66.3% and 67% of the maximum proanthocyanidins, ABTS, DPPH and CUPRAC values, respectively. This study revealed that MAE is a reliable and efficient method for extracting high yields of phytochemicals from E. robusta, with significant potential to be up-scaled for industrial, nutraceutical or pharmaceutical applications.

The main aim of this study was to develop rice starch (RS), ι-carrageenan (ι-car) based film. Different formulations of RS (1-4%, w/w), ι-car (0.5-2%, w/w) was blended with stearic acid (SA; 0.3-0.9%, w/w) and glycerol (1%, w/w) as a plasticizer. The effect of film ingredients on the thickness, water vapour permeability (WVP), film solubility (FS), moisture content (MC), colour, film opacity (FO), tensile strength (TS), elongation-at-break (EAB) of film was examined. Interactions and miscibility of partaking components was studied by using Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). Hydrocolloid suspension solution of mix polysaccharides imparted a significant impact (p<0.05) on the important attributes of resulting edible film. TS and EAB of film were improved significantly (p<0.05) when ι-car was increased in the film matrix. Formulation F1 comprising 2% ι-car, 2% RS, 0.3% SA, Gly 30% w/w and 0.2% surfactant (tween®20) provided film with good physical, mechanical and barrier properties. FT-IR and XRD results reveal that molecular interactions between RS-ι-car have a great impact on the film properties confining the compatibility and miscibility of mixed polysaccharide. Results of the study offers new biodegradable formulation for application on fruit and vegetables.

<h3>Objective</h3> Pancreatic cancer is a leading cause of cancer-related death in the Western world. Current chemotherapy regimens have modest survival benefit. Thus, novel, effective therapies are required for treatment of this disease. <h3>Design</h3> Activating <i>KRAS</i> mutation almost always drives pancreatic tumour initiation, however, deregulation of other potentially druggable pathways promotes tumour progression. PTEN loss leads to acceleration of <i>Kras^G12D</i>-driven pancreatic ductal adenocarcinoma (PDAC) in mice and these tumours have high levels of mammalian target of rapamycin (mTOR) signalling. To test whether these KRAS PTEN pancreatic tumours show mTOR dependence, we compared response to mTOR inhibition in this model, to the response in another established model of pancreatic cancer, KRAS P53. We also assessed whether there was a subset of pancreatic cancer patients who may respond to mTOR inhibition. <h3>Results</h3> We found that tumours in KRAS PTEN mice exhibit a remarkable dependence on mTOR signalling. In these tumours, mTOR inhibition leads to proliferative arrest and even tumour regression. Further, we could measure response using clinically applicable positron emission tomography imaging. Importantly, pancreatic tumours driven by activated KRAS and mutant p53 did not respond to treatment. In human tumours, approximately 20% of cases demonstrated low PTEN expression and a gene expression signature that overlaps with murine KRAS PTEN tumours. <h3>Conclusions</h3> KRAS PTEN tumours are uniquely responsive to mTOR inhibition. Targeted anti-mTOR therapies may offer clinical benefit in subsets of human PDAC selected based on genotype, that are dependent on mTOR signalling. Thus, the genetic signatures of human tumours could be used to direct pancreatic cancer treatment in the future.

Extracellular vesicles (EVs) are produced and secreted from most cells of the body and can be recovered in biological fluids. Although there has been extensive characterisation of the protein and nucleic acid component of EVs, their lipidome has received little attention and may represent a unique and untapped source of biomarkers for prostate cancer diagnosis and prognosis. EVs were isolated from non-tumourigenic (RWPE1), tumourigenic (NB26) and metastatic (PC-3) prostate cell lines. Lipids were extracted and subsequently used for targeted lipidomics analysis for the quantitation of molecular lipid species. A total of 187 molecular lipid species were quantitatively identified in EV samples showing differential abundance between RWPE1, NB26 and PC-3 EV samples. Fatty acids, glycerolipids and prenol lipids were more highly abundant in EVs from non-tumourigenic cells, whereas sterol lipids, sphingolipids and glycerophospholipids were more highly abundant in EVs from tumourigenic or metastatic cells. This study identified differences in the molecular lipid species of prostate cell-derived EVs, increasing our understanding of the changes that occur to the EV lipidome during prostate cancer progression. These differences highlight the importance of characterising the EV lipidome, which may lead to improved diagnostic and prognostic biomarkers for prostate cancer.

Summary The aim of this study was to investigate the effect of freeze‐drying, hot air‐drying and vacuum‐drying at 70, 90 and 110 °C, on dried lemon pomace polyphenols and antioxidant capacity. The total phenolic content and antioxidant capacity were higher in lemon pomace dried by hot air or under vacuum than those dried by freeze‐drying and increased as the temperature increased. The highest total flavonoid content was recorded in the pomace dried under vacuum at 70 and 90 °C. Lemon pomace dried by freeze‐drying had the highest neohesperidin content, whereas pomace dried under vacuum at 70 °C had the highest rutin and p ‐coumaric acid content. The highest gallic acid content was recorded in the pomace dried by hot air at 110 °C. The results of this study indicate that drying technique should be carefully selected according to the bioactive compounds aimed to be extracted.

The effect of different combinations of maltodextrin (MD) coating agents (MD, MD + soybean protein, and MD + ι-carrageenan) on the encapsulation of lemon by-product aqueous extracts using freeze-drying and spray-drying were investigated. The total phenolic content (TPC), total flavonoid content (TFC), and ferric ion reducing antioxidant power (FRAP) of the microparticles were evaluated. Freeze-drying with the mixture of MD + soybean protein resulted in the highest retention of TPC, TFC, and FRAP (1.66 ± 0.02 mg GAE/g d.b., 0.43 ± 0.02 mg CE/g d.b., and 3.70 ± 0.05 mM TE/g, respectively). Freeze-drying resulted in microparticles with lower moisture content (MC) and water activity (aw) than those produced by spray-drying. Specifically, the MC and aw of the microparticles produced by freeze-drying ranged from 1.15 to 2.15% and 0.13 to 0.14, respectively, while the MC and aw of the microparticles produced by spray-drying ranged from 6.06% to 6.60% and 0.33 to 0.40, respectively. Scanning electron microscopy revealed that spray-drying resulted in the formation of spherical particles of different sizes regardless of the type of coating agent. Although freeze-drying resulted in microparticles with amorphous glassy shapes, the mixture of MD + soybean protein resulted in the formation of spherical porous particles. X-ray diffraction revealed a low degree of crystallinity for the samples produced by both techniques.

The study investigated the possibility of enhancing the shelf life of plum fruit coated with rice starch-ι-carrageenan (RS-ι-car) composite coating blended with sucrose fatty acid esters (FAEs). Film solution (starch 3%, carrageenan 1.5% and FAEs 2%) was prepared by mixing the ingredients and properties of stand-alone films (physical, mechanical, barrier and surface morphology) were studied before applying the coating on fruit surface. Fruit were stored at 20 °C for 3 weeks and analyzed for weight loss, ethylene production, respiration rate, color change, firmness, and titratable acidity (TA) and soluble solid content (SSC). Surface morphology of stand-alone film and fruit surface (after applying on the plum fruit) was studied using scanning electron microscopy (SEM). Phytochemical analysis was performed during the storage period and total phenolic content (TPC), total antioxidant capacity (TAC), flavonoid content (FC) and free radical scavenging activity were determined. The rice starch composite coating was shown to be effective in reducing both weight loss (WL) and respiration rate and inhibiting the endogenous ethylene production when compared to the uncoated control fruit stored at room temperature (p < 0.05). TPC, TAC, FC and free radical scavenging activity was unaffected in the coated fruit throughout the storage period (p < 0.05). The findings reported in this study indicate that the RS-ι-car-FAEs coating prolongs the shelf life and maintains the overall quality of plum fruit during storage and could potentially be commercialized as a new edible coating for the plum fruit industry.

Olive pomace is a waste produced by the olive oil industry in massive quantities each year. Disposal of olive pomace is difficult due to high concentrations of phenolic compounds, which is an environmental concern. However, phenolic compounds have applications in the health industry. Therefore, extraction of phenolic compounds from olive pomace has the potential to remove an environmentally hazardous portion of pomace while creating an additional source of income for farmers and producers. Using advanced technologies including Ultrasound Assisted Extraction (UAE), combined with water as an extraction solvent, has recently gained popularity. The present study outlines the optimal UAE conditions for the extraction of phenolic compounds with high antioxidant activity from olive pomace. Optimal conditions were developed using RSM for parameters power, time and sample-to-solvent ratio. Total phenolic compounds determined by Folin Ciocalteu method and total major bioactive compounds determined by HPLC as well as antioxidant capacity (DPPH and CUPRAC) were investigated. The optimal conditions for the extraction of phenolic compounds with high antioxidant activity were 2 g of dried pomace/100 mL of water at 250 W power for 75 min. UAE improved the extraction efficiency of water and yielded extracts with high levels of phenolic compounds and strong antioxidant activity.

Xao tam phan (Paramignya trimera (Oliv.) Guillaum) is a Vietnamese traditionally medicinal plant used in the treatment of numerous cancers. The preparation of Xao tam phan extracts including solvent type and extraction method have significant effects on extraction efficiency, phytochemical profile and biological activity. This study aimed to investigate the effects of five various solvents (water, acetonitrile, methanol, ethyl acetate and hexane) and three different extraction methods (conventional, ultrasound-assisted and microwave-assisted) on phytochemical yield and antioxidant capacity of P. trimera root from Vietnam. The results indicate that methanol extracted the maximal yield of phytochemicals from P. trimera and exhibited the greatest antioxidant capacity, with eleven compounds were identified and quantified. Microwave-assisted extraction produced the maximal phytochemical yields (except for total flavonoids) and antioxidant capacity, when compared to conventional and ultrasound-assisted extractions. These data reveal that the use of methanol and microwave-assisted extraction are recommended for extraction of biologically active phytochemicals from P. trimera root for application in the nutraceutical and/or pharmaceutical industries.

Current chemotherapy drugs for pancreatic cancer only offer an increase in survival of up to six months. Additionally, they are highly toxic to normal tissues, drastically affecting the quality of life of patients. Therefore, the search for novel agents, which induce apoptosis in cancer cells while displaying limited toxicity towards normal cells, is paramount. The olive biophenols, oleuropein, hydroxytyrosol and tyrosol, have displayed cytotoxicity towards cancer cells without affecting non-tumorigenic cells in cancers of the breast and prostate. However, their activity in pancreatic cancer has not been investigated. Therefore, the aim of this study was to determine the anti-pancreatic cancer potential of oleuropein, hydroxytyrosol and tyrosol. Pancreatic cancer cells (MIA PaCa-2, BxPC-3, and CFPAC-1) and non-tumorigenic pancreas cells (HPDE) were treated with oleuropein, hydroxytyrosol and tyrosol to determine their effect on cell viability. Oleuropein displayed selective toxicity towards MIA PaCa-2 cells and hydroxytyrosol towards MIA PaCa-2 and HPDE cells. Subsequent analysis of Bcl-2 family proteins and caspase 3/7 activation determined that oleuropein and hydroxytyrosol induced apoptosis in MIA PaCa-2 cells, while oleuropein displayed a protective effect on HPDE cells. Gene expression analysis revealed putative mechanisms of action, which suggested that c-Jun and c-Fos are involved in oleuropein and hydroxytyrosol induced apoptosis of MIA PaCa-2 cells.

Background & AimsCurrent methods of preoperative staging and predicting outcome following pancreatectomy for pancreatic cancer (PC) are inadequate. We evaluated the utility of multiple biomarkers from distinct biologic pathways as potential predictive markers of response to pancreatectomy and patient survival.MethodsWe assessed the relationship of candidate biomarkers known, or suspected, to be aberrantly expressed in PC, with disease-specific survival and response to therapy in a cohort of 601 patients.ResultsOf the 17 candidate biomarkers examined, only elevated expression of S100A2 was an independent predictor of survival in both the training (n = 162) and validation sets (n = 439; hazard ratio [HR], 2.19; 95% confidence interval [CI]: 1.48–3.25; P < .0001) when assessed in a multivariate model with clinical variables. Patients with high S100A2 expressing tumors had no survival benefit with pancreatectomy compared with those with locally advanced disease, whereas those without high S100A2 expression had a survival advantage of 10.6 months (19.4 vs 8.8 months, respectively) and a HR of 3.23 (95% CI: 2.39–4.33; P < .0001). Of significance, patients with S100A2-negative tumors had a significant survival benefit from pancreatectomy even in the presence of involved surgical margins (median, 15.7 months; P = .0007) or lymph node metastases (median, 17.4 months; P = .0002).ConclusionsS100A2 expression is a good predictor of response to pancreatectomy for PC and suggests that high S100A2 expression may be a marker of a metastatic phenotype. Prospective measurement of S100A2 expression in diagnostic biopsy samples has potential clinical utility as a predictive marker of response to pancreatectomy and other therapies that target locoregional disease. Current methods of preoperative staging and predicting outcome following pancreatectomy for pancreatic cancer (PC) are inadequate. We evaluated the utility of multiple biomarkers from distinct biologic pathways as potential predictive markers of response to pancreatectomy and patient survival. We assessed the relationship of candidate biomarkers known, or suspected, to be aberrantly expressed in PC, with disease-specific survival and response to therapy in a cohort of 601 patients. Of the 17 candidate biomarkers examined, only elevated expression of S100A2 was an independent predictor of survival in both the training (n = 162) and validation sets (n = 439; hazard ratio [HR], 2.19; 95% confidence interval [CI]: 1.48–3.25; P < .0001) when assessed in a multivariate model with clinical variables. Patients with high S100A2 expressing tumors had no survival benefit with pancreatectomy compared with those with locally advanced disease, whereas those without high S100A2 expression had a survival advantage of 10.6 months (19.4 vs 8.8 months, respectively) and a HR of 3.23 (95% CI: 2.39–4.33; P < .0001). Of significance, patients with S100A2-negative tumors had a significant survival benefit from pancreatectomy even in the presence of involved surgical margins (median, 15.7 months; P = .0007) or lymph node metastases (median, 17.4 months; P = .0002). S100A2 expression is a good predictor of response to pancreatectomy for PC and suggests that high S100A2 expression may be a marker of a metastatic phenotype. Prospective measurement of S100A2 expression in diagnostic biopsy samples has potential clinical utility as a predictive marker of response to pancreatectomy and other therapies that target locoregional disease.

Myc oncoproteins and histone deacetylases (HDACs) modulate gene transcription and enhance cancer cell proliferation, and HDAC inhibitors are among the most promising new classes of anticancer drugs. Here, we show that N-Myc and c-Myc upregulated HDAC2 gene expression in neuroblastoma and pancreatic cancer cells, respectively, which contributed to N-Myc- and c-Myc-induced cell proliferation. Cyclin G2 (CCNG2) was commonly repressed by N-Myc and HDAC2 in neuroblastoma cells and by c-Myc and HDAC2 in pancreatic cancer cells, and could be reactivated by HDAC inhibitors. 5-bromo-2′-deoxyuridine incorporation assays showed that transcriptional repression of CCNG2 was, in part, responsible for N-Myc-, c-Myc- and HDAC2-induced cell proliferation. Dual crosslinking chromatin immunoprecipitation assay demonstrated that N-Myc acted as a transrepressor by recruiting the HDAC2 protein to Sp1-binding sites at the CCNG2 gene core promoter. Moreover, HDAC2 was upregulated, and CCNG2 downregulated, in pre-cancerous and neuroblastoma tissues from N-Myc transgenic mice, and c-Myc overexpression correlated with upregulation of HDAC2 and repression of CCNG2 in tumour tissues from pancreatic cancer patients. Taken together, our data indicate the critical roles of upregulation of HDAC2 and suppression of CCNG2 in Myc-induced oncogenesis, and have significant implications for the application of HDAC inhibitors in the prevention and treatment of Myc-driven cancers.

Phyllanthus amarus (P. amarus) has been used as a herbal medicine, particularly for liver support, in many countries and its extracts have been shown to possess potent antioxidant and anticancer properties in vitro. The preparation of dried sample is crucial for further extraction and isolation of phytochemicals. In this study, the effects of six different drying methods (hot air, low-temperature air, infrared, microwave, sun, and vacuum drying) on the phytochemical yield and antioxidant capacity were determined to identify the optimal drying method for P. amarus. The results showed that different drying methods, as well as different drying conditions within each method, significantly affected phytochemical yield and antioxidant capacity of P. amarus extracts. Infrared drying at 30°C was the best method for both retention of bioactive compound yield and antioxidant capacity of P. amarus extract, with 12 compounds were identified. In contrast, low-temperature-air drying at 25°C not only required the longest drying time but also significantly reduced the levels of bioactive compounds and antioxidant capacity of P. amarus. Therefore, infrared drying at 30°C is suggested for drying P. amarus for subsequent assessment of bioactivity.

Pancreatic cancer has a dismal prognosis and is the fourth most common cause of cancer related death in Western societies. In large part this is due to its typically late presentation, usually as locally advanced or metastatic disease. Identification of the non-invasive precursor lesions to pancreatic cancer raises the possibility of surgical treatment or chemoprevention at an early stage in the evolution of this disease, when more amenable to therapeutic interventions. Precursor lesions to pancreatic ductal adenocarcinoma, in particular pancreatic intraepithelial neoplasia (PanIN), have been recognised under a variety of synonyms for over 50 years. Over the past decade our understanding of the morphology, biological significance and molecular aberrations of these lesions has grown rapidly and there is now a widely accepted progression model integrating the accumulated morphological and molecular observations. Further progress is likely to be accelerated by improved mouse models of pancreatic cancer and by insight into the cancer genome gained by the International Cancer Genome Consortium (ICGC), in which an Australian consortium is leading the pancreatic cancer initiative. This review also outlines the morphological and molecular features of the other two precursors of pancreatic ductal adenocarcinoma, i.e., intraductal papillary mucinous neoplasms and mucinous cystic neoplasms.

Active food packaging based on pea starch and guar gum (PSGG) films containing natural antioxidants (NAs) was developed. Four kinds of NAs (epigallocatechin gallate (EGCG), blueberry ash (BBA) fruit extract, macadamia (MAC) peel extract, and banana (BAN) peel extract) were added into the PSGG-based films as antioxidant additive. The effects of these compounds at different amounts on the physical and antioxidant characteristics of the PSGG film were investigated. The antioxidant activity was calculated with three analytical assays: DPPH radical scavenging ability assay, cupric reducing antioxidant capacity (CUPRAC), and ferric reducing activity power (FRAP). EGCG-PSGG films showed higher antioxidant activity, followed by BBA-PSGG, MAC-PSGG, and BAN-PSGG films, at all concentrations (0.75–3 mg/mL) and with all procedures tested. Additionally, the antioxidant activity of films showed a concentration dependency. The results revealed that addition of NAs made the PSGG film darker and less transparent. However, the moisture barrier was significantly improved when NAs were incorporated into the film. The FTIR spectra were examined to determine the interactions between polymers and NAs. The results suggested that incorporation of EGCG, BBA, MAC, and BAN into PSGG films have great potential for use as active food packaging for food preservation.

Adjuvant chemotherapy improves survival for patients with resected pancreatic cancer. Elderly patients are under-represented in Phase III clinical trials, and as a consequence the efficacy of adjuvant therapy in older patients with pancreatic cancer is not clear. We aimed to assess the use and efficacy of adjuvant chemotherapy in older patients with pancreatic cancer.We assessed a community cohort of 439 patients with a diagnosis of pancreatic ductal adenocarcinoma who underwent operative resection in centres associated with the Australian Pancreatic Cancer Genome Initiative.The median age of the cohort was 67 years. Overall only 47% of all patients received adjuvant therapy. Patients who received adjuvant chemotherapy were predominantly younger, had later stage disease, more lymph node involvement and more evidence of perineural invasion than the group that did not receive adjuvant treatment. Overall, adjuvant chemotherapy was associated with prolonged survival (median 22.1 vs 15.8 months; P<0.0001). Older patients (aged ≥70) were less likely to receive adjuvant chemotherapy (51.5% vs 29.8%; P<0.0001). Older patients had a particularly poor outcome when adjuvant therapy was not delivered (median survival=13.1 months; HR 1.89, 95% CI: 1.27-2.78, P=0.002).Patients aged ≥70 are less likely to receive adjuvant therapy although it is associated with improved outcome. Increased use of adjuvant therapy in older individuals is encouraged as they constitute a large proportion of patients with pancreatic cancer.

Summary Six brown algae, Sargassum vestitum, Sargassum linearifolium, Phyllospora comosa, Padina sp. , Hormosira banksii and Sargassum podocanthum, were investigated for the chemical profile and antioxidant activity. The results showed that the extracts H. banksii, S. vestitum and Padina sp. indicated the significantly higher total phenolic compound ( TPC ) and antioxidant activities ( ABTS , DPPH and FRAP ) compared to the other species ( P &lt; 0.05) and comparable to positive controls: butylated hydroxytoluene, ascorbic acid and alpha‐tocopherol at the concentrations (0.06–1 mg mL −1 ). Fucoxanthin was also found in six species and isolated for evaluating antioxidant activity. In addition, the phenolic compounds were mainly responsible for antioxidant activity of the extracts, while fucoxanthin showed quite high antioxidant activity. It is suggested that S. vestitum, H. banksii and Padina sp. have the potent for extracting bioactive components and further applications in food and pharmaceutical industries.

Euphorbia tirucalli is a succulent shrub or small tree that is native to the African continent, however, it is widely cultivated across the globe due to its use in traditional medicines to treat ailments, ranging from scorpion stings to HIV. Recent studies have identified compounds present in the latex of the plant, including a range of bi- and triterpenoids that exhibit bioactivity, including anticancer activity. This study aimed to optimize water extraction conditions for high-yield total phenolic content recovery, to prepare methanol and aqueous extracts from the aerial sections of the plant, and to test the phytochemical, antioxidant, and anti-cancer properties of these extracts. Water extraction of total phenolic compounds (TPC) was optimized across a range of parameters including temperature, extraction time, and plant mass-to-solvent ratio. The water extract of the E. tirucalli powder was found to contain TPC of 34.01 mg GAE (gallic acid equivalents)/g, which was approximately half that of the methanol extract (77.33 mg GAE/g). The results of antioxidant assays showed a uniform trend, with the methanol extract's antioxidant reducing activity exceeding that of water extracts, typically by a factor of 2:1. Regression analysis of the antioxidant assays showed the strongest correlation between extract TPC and antioxidant activity for the ABTS (2,2-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) methods. The methanol extract also showed greater growth inhibition capacity towards the MiaPaCa-2 pancreatic cancer cell line. These data suggest that further investigations are required to confirm the source of activity within the E. tirucalli leaf and stems for potential use in the nutraceutical and pharmaceutical industries.

The genus Eucalyptus (Myrtaceae) is mainly native to Australia; however, some species are now distributed globally. Eucalyptus has been used in indigenous Australian medicines for the treatment of a range of aliments including colds, flu, fever, muscular aches, sores, internal pains, and inflammation. Eucalyptus oils containing volatile compounds have been widely used in the pharmaceutical and cosmetics industries for a multitude of purposes. In addition, Eucalyptus extracts containing nonvolatile compounds are also an important source of key bioactive compounds, and several studies have linked Eucalyptus extracts with anticancer properties. With the increasing research interest in Eucalyptus and its health properties, this review briefly outlines the botanical features of Eucalyptus, discusses its traditional use as medicine, and comprehensively reviews its phytochemical and anticancer properties and, finally, proposes trends for future studies.

Myc oncoproteins exert tumorigenic effects by regulating expression of target oncogenes. Histone H3 lysine 79 (H3K79) methylation at Myc-responsive elements of target gene promoters is a strict prerequisite for Myc-induced transcriptional activation, and DOT1L is the only known histone methyltransferase that catalyzes H3K79 methylation. Here, we show that N-Myc upregulates DOT1L mRNA and protein expression by binding to the DOT1L gene promoter. shRNA-mediated depletion of DOT1L reduced mRNA and protein expression of N-Myc target genes ODC1 and E2F2 DOT1L bound to the Myc Box II domain of N-Myc protein, and knockdown of DOT1L reduced histone H3K79 methylation and N-Myc protein binding at the ODC1 and E2F2 gene promoters and reduced neuroblastoma cell proliferation. Treatment with the small-molecule DOT1L inhibitor SGC0946 reduced H3K79 methylation and proliferation of MYCN gene-amplified neuroblastoma cells. In mice xenografts of neuroblastoma cells stably expressing doxycycline-inducible DOT1L shRNA, ablating DOT1L expression with doxycycline significantly reduced ODC1 and E2F2 expression, reduced tumor progression, and improved overall survival. In addition, high levels of DOT1L gene expression in human neuroblastoma tissues correlated with high levels of MYCN, ODC1, and E2F2 gene expression and independently correlated with poor patient survival. Taken together, our results identify DOT1L as a novel cofactor in N-Myc-mediated transcriptional activation of target genes and neuroblastoma oncogenesis. Furthermore, they characterize DOT1L inhibitors as novel anticancer agents against MYCN-amplified neuroblastoma. Cancer Res; 77(9); 2522-33. ©2017 AACR.

The aim of this study was to develop an optimal formulation for preparation of edible films from chitosan, pea starch and glycerol using response surface methodology. Three independent variables were assigned comprising chitosan (1-2%), pea starch (0.5-1.5%) and glycerol (0.5-1%) to design an empirical model best fit in physical, mechanical and barrier attributes. Impacts of independent variables on thickness, moisture content, solubility, tensile strength, elastic modulus, elongation at break and water vapor permeability of films were evaluated. All the parameters were found to have significant effects on physical and mechanical properties of film. The optimal formulation for preparation of edible film from chitosan, pea starch and glycerol was 1% chitosan, 1.5% pea starch and 0.5% glycerol. Edible films with good physical and mechanical properties can be prepared with this formulation and thus this formulation can be further applied for testing on coating for fruit and vegetables.

Polyphenols of citrus by-products, due to their antioxidant and antimicrobial activities, could be valorized by pharmaceutical and food industries, adding a value to the citrus processing companies. A number of studies have investigated the effect of ultrasound-assisted extraction (UAE) conditions on the recovery of phenolics derived from citrus waste using both organic solvents or mixed aqueous solvent systems. To maximize efficiency, UAE conditions should be tailored to the physical parameters of the solvent(s) employed. The aim of this study was to investigate the effect of four UAE parameters: particle size (1.40–2.80 mm), extraction time (10–60 min), extraction temperature (23–50 °C) and ultrasonic power (150–250 W) on the simultaneous recovery of p-coumaric acid, caffeic acid, chlorogenic acid, and hesperidin from citrus waste using pure water as a solvent. High-performance liquid chromatography (HPLC) was employed for the identification and quantification of the cited compounds. Particle size was determined to be an important parameter affecting compound recovery, with the exception of chlorogenic acid. A particle size of 1.40 mm resulted in the highest recovery of p-coumaric and caffeic acids (0.25 and 0.58 mg/g, respectively), while higher hesperidin yields were achieved from the particle sizes of 2.00 and 1.40 mm (6.44 and 6.27 mg/g, respectively). Extraction temperature significantly affected only the recovery of the flavanone glycoside (P < 0.05). As the extraction temperature increased from 30 to 50 °C the recovery of hesperidin increased from 6.59 to 7.84 mg/g, respectively. Neither extraction time nor ultrasonic power significantly affected the recovery of any individual phenolic compound.

Olea europaea L. leaves are an agricultural waste product with a high concentration of phenolic compounds; especially oleuropein. Oleuropein has been shown to exhibit anti-proliferative activity against a number of cancer types. However, they have not been tested against pancreatic cancer, the fifth leading cause of cancer related death in Western countries. Therefore, water, 50% ethanol and 50% methanol extracts of Corregiola and Frantoio variety Olea europaea L. leaves were investigated for their total phenolic compounds, total flavonoids and oleuropein content, antioxidant capacity and anti-proliferative activity against MiaPaCa-2 pancreatic cancer cells. The extracts only had slight differences in their phytochemical properties, and at 100 and 200 μg/mL, all decreased the viability of the pancreatic cancer cells relative to controls. At 50 μg/mL, the water extract from the Corregiola leaves exhibited the highest anti-proliferative activity with the effect possibly due to early eluting HPLC peaks. For this reason, olive leaf extracts warrant further investigation into their potential anti-pancreatic cancer benefits.

The mechanical properties and moisture sorption at relative humidity (RH) range of 11–94%, water vapor permeability (WVP), solubility in water and color of the pea starch films as a function of glycerol were examined. The results showed that increasing the concentration of plasticizer resulted in improvement of the tensile strength of the films at RH <43%, the percent elongation as well as the deformation at break at RH <84%. Increasing plasticizer content and RH also resulted in films with lower Young's modulus, lower puncture force, but higher puncture deformation. Furthermore, increasing plasticizer content led to the films with more opaque appearance. Films prepared with 15 and 25% glycerol had lower WVP in comparison with unplasticized film. This study provides information regarding the advantageous or disadvantageous of possible application of pea starch films in food packaging industry. Practical Application Starch edible films have been utilized for packaging technologies and edible coatings. Pea starch has been found to produce the films with improved physical and mechanical properties in comparison with films prepared from other starches due to high amount of amylose. The development of pea starch film with improved functions affects its application. Pea starch edible films may find practical applications in the poultry, meat, seafood, fruit, vegetable, grains and candies industries.

Antimicrobial activity of epigallocatechin-3-gallate (EGCG) and two native Australian plants blueberry ash (BBA) fruit and macadamia (MAC) skin extracts against nine pathogenic and spoilage bacteria and seven strains of fungi, using an agar well diffusion assay were investigated. The minimum inhibitory concentrations (MIC) of these compounds were calculated using 96-well microtiter plates method. Finally, active antimicrobial packaging films were prepared by incorporation of EGCG, BBA and MAC extracts at 1-, 2-, 3-, and 4-fold of their correspondence MIC values into edible films based on pea starch and guar gum (PSGG). The antimicrobial activity of films was investigated against target microorganisms by agar disc diffusion technique and quantified using the viable cell count assay. Among the test microorganisms, Salmonella typhimurium and Rhizopus sp. were the most resistance to active films. Films containing EGCG showed the highest activity against all test strains. As the concentration of compounds increased higher than 2 × MIC, the mechanical characteristics of the films were affected considerably. The results indicated that EGCG-PSGG, BBA-PSGG and MAC-PSGG films can be used as active food packaging systems for preserving food safety and prolonging the shelf-life of the packaged food.

Improvements in the hygroscopic properties of starch based films are important to strengthen their mechanical properties. The effects of different hydrophobic components-butyric acid (BA, C4:0), lauric acid (LA, C12:0), palmitic acid (PA, C16:0), oleic acid (OA, C18:1), stearic acid (SA, C18:0) and sucrose fatty acid ester (FAEs) on the rice starch (RS)-ι-carrageenan (ι-car) composite films were investigated. Scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) in combination with amylose-lipid complexing index (CI) were used to characterise the changes in structure and properties of edible films. The SEM results showed that the surface of films became smoother after the incorporation of fatty acids. Carbon-chain length was a major determinant of CI formation which further influenced the attributes of RS-ι-car films. The addition of FAEs to RS-ι-car improved film thickness, permeability, transparency, tensile properties (TS) and could be used to tailor biodegradable edible films with enhanced properties and future fruit coating applications.

To facilitate intercellular communication, cells release nano-sized, extracellular vesicles (EVs) to transfer biological cargo to both local and distant sites. EVs are enriched in tetraspanins, two of which (CD9 and CD151) have altered expression patterns in many solid tumours, including prostate cancer, as they advance toward metastasis. We aimed to determine whether EVs from prostate cells with altered CD9 and CD151 expression could influence cellular behaviour and increase the metastatic capabilities of non-tumourigenic prostate cells. EVs were isolated by ultrafiltration and characterised for their tetraspanin expression and size distribution. iTRAQ was used to identify differences between RWPE1 and tetraspanin-modified RWPE1 EV proteomes, showing an enrichment in protein degradation pathways. Addition of EVs from RWPE1 cells with reduced CD9 or increased CD151 abundance resulted in increased invasion of RWPE1 cells, and increased migration in the case of high CD151 abundance. We have been able to show that alteration of CD9 and CD151 on prostate cells alters the proteome of their resultant EVs, and that these EVs can enhance the migratory and invasive capabilities of a non-tumourigenic prostate cellular population. This work suggests that cellular tetraspanin levels can alter EVs, potentially acting as a driver of metastasis in prostate cancer.

A significant amount of banana peels is generated as waste annually and shows great potential as a lead material for further utilization in the nutraceutical industry. However, potentiality of banana peel utilization largely depends on the favorable drying condition of the material before it can be used for further processing. Therefore, it is necessary to identify the suitable drying conditions for banana peel. This study investigated the effect of six different drying methods on the quality of banana peels. The results showed that different drying conditions significantly affected the physical, chemical, and antioxidant properties of dried peels. Microwave irradiation at the power level of 960 W for 6 min was the most suitable condition, as these dried peels had good physical properties, minimum loss of bioactive compounds, and antioxidant properties. This was followed by freeze-drying, vacuum oven at 60°C, hot air oven at 120°C, dehumidified air at 60°C, and sun drying. The peels dried by microwave possessed a total phenolic content of 25.26 mg of gallic acid equivalents/g of dry matter (DM) and potent antioxidant capacity [(1,1-diphenyl-2-picrylhydrazyl) of 37.70; 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid of 46.35; ferric reducing antioxidant power of 45.94; and cupric ion reducing antioxidant capacity of 64.55 mg of trolox equivalents/g of DM]. Therefore, the study recommends the use of microwave irradiation under the studied condition (power level of 960 W for 6 min) for further processing and utilization.

Response surface methodology (RSM) based on a three-factor and three-level Box–Behnken design was employed for optimizing the aqueous ultrasound-assisted extraction (AUAE) conditions, including extraction time (35–45 min), extraction temperature (45–55 °C) and ultrasonic power (150–250 W), for the recovery of total phenolic content (TPC) and rutin from lemon by-products. The independent variables and their values were selected on the basis of preliminary experiments, where the effects of five extraction parameters (particle size, extraction time and temperature, ultrasonic power and sample-to-solvent ratio) on TPC and rutin extraction yields were investigated. The yields of TPC and rutin were studied using a second-order polynomial equation. The optimum AUAE conditions for TPC were extraction time of 45 min, extraction temperature of 50 °C and ultrasonic power of 250 W with a predicted value of 18.10 ± 0.24 mg GAE/g dw, while the optimum AUAE conditions for rutin were extraction time of 35 min, extraction temperature of 48 °C and ultrasonic power of 150W with a predicted value of 3.20 ± 0.12 mg/g dw. The extracts obtained at the optimum AUAE conditions were compared with those obtained by a hot water and an organic solvent conventional extraction in terms of TPC, total flavonoid content (TF) and antioxidant capacity. The extracts obtained by AUAE had the same TPC, TF and ferric reducing antioxidant power as those achieved by organic solvent conventional extraction. However, hot water extraction led to extracts with the highest flavonoid content and antioxidant capacity. Scanning electron microscopy analysis showed that all the extraction methods led to cell damage to varying extents.

Banana peel is a by-product from the food industry, which is rich in dietary fiber and phenolic compounds. It is hypothesised that physicochemical and antioxidant properties of banana peel are significantly changed through different ripening stages. Ethylene treatment for fastening ripening is presumed to influence the changes in physicochemical and antioxidant properties of the peel. As such, this study investigated the impact of ripening stages with and without ethylene treatments on the changes of phytochemicals and antioxidant properties of banana peel. Green bananas were ripened with ethylene (1 ppm and 16 ppm) and without ethylene. The peel was then evaluated for the colour changes, chlorophylls, carotenoids, flavonoids, proanthocyanidins, total phenolic contents, and antioxidants capacity. As the fruit colour changed from green to yellow, chlorophyll was degraded by approximately 90%, while carotenoids and flavonoids increased by approximately 50% and 27%, respectively. In addition, levels of phenolics, proanthocyanidins, and antioxidant capacity also increased as the fruit continued to ripen. However, the overripe fruit peel lost up to 21% of its antioxidant capacity and up to 44% of its phytochemicals. Though statistically not significant, the peel without ethylene treatment had a relatively higher phenolic content and antioxidant power than those of the peel treated by ethylene at stages 5–7. Overall, the banana peel at ripening stages 5–7 ha s the highest levels of physicochemical and antioxidant properties and thus the peel at these stages is recommended for recovery of phenolic compounds for further applications. Additionally, the PCA biplot shows that the ripening stages could be characterised by the clustering behaviour of certain phytochemicals. Therefore, it is suggested to consider phytochemical changes when evaluating the ripening stages of banana.

Abstract Background and Aim: Clinicopathological data regarding pancreatic solid pseudopapillary tumors (SPT) in a multiethnic country are limited. The aim of the present study was to characterize pancreatic SPT in Australia. Methods: Clinicopathological features, treatment, immunohistochemical findings and outcome data of 34 patients (79% Caucasian, 12% Asian, 6% South Pacific Islander and 3% African) with pancreatic SPT were reviewed. Results: The most presenting complaint was abdominal pain. Median diameter of tumors was 60 mm (range: 20–220); predominantly located in the pancreatic tail (tail : body : head = 23:3:8). All tumors were resected and patients underwent surgery, including a liver resection for metastasis, all patients were alive after a median follow up of 70 months (IQR: 48–178). Two patients underwent repeated surgery for local recurrences with liver metastases after 8 and 18 months, which were successfully managed by surgical resection. Completeness of excision, perineural spread, vascular space invasion, mitotic rate and cellular atypia did not predict recurrence. In all cases, there was aberrant nuclear staining of beta‐catenin and a loss of membranous expression of E‐cadherin with aberrant nuclear localization of the cytoplasmic domain. Most pancreatic SPT were also strongly positive for CD10 (96%), progesterone receptor (79%), cytokeratin (28%), synapthophysin (26%) and chromogranin (15%). Conclusions: Pancreatic SPT occur in all races and are uniformly indolent. Given complete resection of a pancreatic SPT is usually curative and recurrences can be treated with re‐operation, correct diagnosis is important.

Chronic pancreatitis and pancreatic cancer are characterised by extensive stellate cell mediated fibrosis, and current therapeutic development includes targeting pancreatic cancer stroma and tumor-host interactions. Recent evidence has suggested that circulating bone marrow derived stem cells (BMDC) contribute to solid organs. We aimed to define the role of circulating haematopoietic cells in the normal and diseased pancreas.Whole bone marrow was harvested from male β-actin-EGFP donor mice and transplanted into irradiated female recipient C57/BL6 mice. Chronic pancreatitis was induced with repeat injections of caerulein, while carcinogenesis was induced with an intrapancreatic injection of dimethylbenzanthracene (DMBA). Phenotype of engrafted donor-derived cells within the pancreas was assessed by immunohistochemistry, immunofluorescence and in situ hybridisation.GFP positive cells were visible in the exocrine pancreatic epithelia from 3 months post transplantation. These exhibited acinar morphology and were positive for amylase and peanut agglutinin. Mice administered caerulein developed chronic pancreatitis while DMBA mice exhibited precursor lesions and pancreatic cancer. No acinar cells were identified to be donor-derived upon cessation of cerulein treatment, however rare occurrences of bone marrow-derived acinar cells were observed during pancreatic regeneration. Increased recruitment of BMDC was observed within the desmoplastic stroma, contributing to the activated pancreatic stellate cell (PaSC) population in both diseases. Expression of stellate cell markers CELSR3, PBX1 and GFAP was observed in BMD cancer-associated PaSCs, however cancer-associated, but not pancreatitis-associated BMD PaSCs, expressed the cancer PaSC specific marker CELSR3.This study demonstrates that BMDC can incorporate into the pancreas and adopt the differentiated state of the exocrine compartment. BMDC that contribute to the activated PaSC population in chronic pancreatitis and pancreatic cancer have different phenotypes, and may play important roles in these diseases. Further, bone marrow transplantation may provide a useful model for the study of tumor-host interactions in cancer and pancreatitis.

Pancreatic cancer is a devastating cancer that presents late, is rapidly progressive and has current therapeutics with only limited efficacy. Bioactive compounds are ubiquitously present in fruits and numerous studies in vitro are addressing the activity of these compounds against pancreatic cancer, thus studies of specific bioactive compounds could lead to new anti-pancreatic cancer strategies. Australian native fruits have been used as foods and medicines by Australian Aboriginals for thousands of years, and preliminary studies have found these fruits to contain rich and diversified bioactive components with high antioxidant activity. Thus, Australian native fruits may possess key components for preventing or delaying the onset of tumorigenesis, or for the treatment of existing cancers, including pancreatic cancer. Numerous databases including PubMed, SciFinder, Web of Knowledge, Scopus, and Sciencedirect were analysed for correlations between bioactive components from fruits and pancreatic cancer, as well as studies concerning Australian native fruits. In this review, we comprehensively highlight the proposed mechanisms of action of fruit bioactives as anti-cancer agents, update the potential anti-pancreatic cancer activity of various major classes of bioactive compounds derived from fruits, and discuss the existence of bioactive compounds identified from a selection Australian native fruits for future studies. Bioactive compounds derived from fruits possess the potential for the discovery of new anti-pancreatic cancer strategies. Further, Australian native fruits are rich in polyphenols including some flora that contain unique phenolic compounds, thereby warranting further investigations into their anti-cancer properties.

Euphol identified in Euphorbia tirucalli (E. tirucalli) has been linked with various health benefits. This study aimed to optimize ultrasonic extraction conditions for euphol from E. tirucalli leaf. Different solvents were tested to determine the most effective solvent for extraction of euphol. Then, response surface methodology (RSM) was employed to optimize ultrasound-assisted extraction conditions including temperature, time and power for maximal extraction of euphol. Our results showed that ethyl acetate:ethanol (4:1, v/v) was the most effective solvent for the extraction of euphol. Ultrasonic temperature and time had a positive impact, whereas, ultrasonic power had a negative effect on the extraction efficiency of euphol. The optimum ultrasonic extraction conditions for euphol were identified as: solvent-to-fresh sample ratio of 100:32 mL/g; ultrasonic temperature of 60 °C; ultrasonic time of 75 min and ultrasonic power of 60% (150 W). Under these optimum conditions, approximately 4.06 mg of euphol could be obtained from one gram of fresh E. tirucalli leaf. This extract also contained phenolic compounds (2.5 mg GAE/g FW) and possessed potent antioxidant capacity. These optimal conditions are applicable for a larger scale to extract and isolate euphol for potential utilization in the pharmaceutical industry.

Summary The papaya ( Carica papaya ) leaf (PL) contains high levels of saponins and polyphenolic compounds, and historically, it has been used as a folk medicine for numerous ailments, including cancer. PL is traditionally prepared by hot water extraction; however, optimised extraction conditions have not been assessed. This study optimised conditions for the extraction of saponins from PL and assessed their antioxidant capacity and antipancreatic cancer activity. Optimisation was achieved using response surface methodology. Saponins and total phenolic compounds were assessed for their antioxidant, free radical scavenging, ion‐reducing capacity, and antipancreatic cancer activity. Optimal aqueous extraction conditions were 85 °C, 25 min. and a water‐to‐leaf ratio of 20:1 mL g −1 . Ethanol extracts demonstrated higher antioxidant, free radical scavenging and ion‐reducing capacity, as well as antipancreatic cancer activity. This study revealed that the PL contains numerous bioactive compounds, with significant anticancer activity warranting further studies on the isolation and characterisation of individual bioactive compounds from the PL.

Olive leaves are an agricultural waste of the olive-oil industry representing up to 10% of the dry weight arriving at olive mills.Disposal of this waste adds additional expense to farmers.Olive leaves have been shown to have a high concentration of phenolic compounds.In an attempt to utilize this waste product for phenolic compounds, we optimized their extraction using water-a "green" extraction solvent that has not yet been investigated for this purpose.Experiments were carried out according to a Box Behnken design, and the best possible combination of temperature, extraction time and sample-to-solvent ratio for the extraction of phenolic compounds with a high antioxidant activity was obtained using RSM; the optimal conditions for the highest yield of phenolic compounds was 90 °C for 70 min at a sample-to-solvent ratio of 1:100 g/mL; however, at 1:60 g/mL, we retained 80% of the total phenolic compounds and maximized antioxidant capacity.Therefore the sample-to-solvent ratio of 1:60 was chosen as optimal and used for further validation.The validation test fell inside the confidence range indicated by the RSM output; hence, the statistical model was trusted.The proposed method is inexpensive, easily up-scaled to industry and shows potential as an additional source of income for olive growers.

A large quantity of banana peel is generated annually and is considered as waste with low value. This study aimed to optimize five extraction parameters; including ultrasonic temperature, time and power, as well as acetone concentration and sample to solvent ratio, for maximum recovery of phenolic compounds, flavonoids, proanthocyanidins and antioxidant properties from banana (Musa cavendish) peel using response surface methodology. The results showed that recovery yields of phenolic compounds, flavonoids, proanthocyanidins and antioxidant properties were affected by the extraction parameters; of which the acetone concentration had the greatest effect. Optimal extraction conditions were found to be at ultrasonic temperature of 30°C, ultrasonic time of 5 min, ultrasonic power of 150 W, sample to solvent ratio of 8:100 g/mL and acetone concentration of 60%. Under these optimal conditions, 23.49 mg of phenolic compounds, 39.46 mg of flavonoids and 13.11 mg of proanthocyanidins could be extracted from 1 g of banana (M. cavendish) peel. Practical applications Banana peel known as waste is generated in a big quantity with limited utilization. Therefore, it is necessary to utilize this by-product for adding value to food industry. This study was designed to establish a simple, effective extraction method for maximum recovery of phenolic compounds from banana peel. Findings from this study can be used for further isolation and purification of phenolic compounds from banana peel for subsequent application in nutraceutical and pharmaceutical industry.

We aimed to define preoperative clinical and molecular characteristics that would allow better patient selection for operative resection.Although we use molecular selection methods for systemic targeted therapies, these principles are not applied to surgical oncology. Improving patient selection is of vital importance for the operative treatment of pancreatic cancer (pancreatic ductal adenocarcinoma). Although surgery is the only chance of long-term survival, 80% still succumb to the disease and approximately 30% die within 1 year, often sooner than those that have unresected local disease.In 3 independent pancreatic ductal adenocarcinoma cohorts (total participants = 1184) the relationship between aberrant expression of prometastatic proteins S100A2 and S100A4 and survival was assessed. A preoperative nomogram based on clinical variables available before surgery and expression of these proteins was constructed and compared to traditional measures, and a postoperative nomogram.High expression of either S100A2 or S100A4 was independent poor prognostic factors in a training cohort of 518 participants. These results were validated in 2 independent patient cohorts (Glasgow, n = 198; Germany, n = 468). Aberrant biomarker expression stratified the cohorts into 3 distinct prognostic groups. A preoperative nomogram incorporating S100A2 and S100A4 expression predicted survival and nomograms derived using postoperative clinicopathological variables.Of those patients with a poor preoperative nomogram score, approximately 50% of patients died within a year of resection. Nomograms have the potential to improve selection for surgery and neoadjuvant therapy, avoiding surgery in aggressive disease, and justifying more extensive resections in biologically favorable disease.

The effects of different solvents on the recovery of (i) extractable solids (ES), (ii) total phenolic compounds (TPC), (iii) total flavonoid content (TFC), (iv) vitamin C, and (v) antioxidant activity from lemon pomace waste were investigated. The results revealed that solvents significantly affected the recovery of ES, TPC, TFC, and antioxidant properties. Absolute methanol and 50% acetone resulted in the highest extraction yields of TPC, whereas absolute methanol resulted in the highest extraction of TFC, and water had the highest recovery of vitamin C. 50% ethanol, and 50% acetone had higher extraction yields for TPC, and TFC, as well as higher antioxidant activity compared with their absolute solvents and water. TPC and TFC were shown to be the major components contributing to the antioxidant activity of lemon pomace.

Banana peel is rich in phenolic compounds and is generally considered as waste. This study aimed to maximise recovery of phenolics from banana peel using water via microwave assisted extraction. The impact of various parameters including pH of solvent, sample to solvent ratio, irradiation time with/without cooling periods, and irradiation power were investigated individually. Following this, extraction conditions were further optimised using Response Surface Methodology. The results revealed that the extraction efficiency can be significantly improved by reducing the pH of water, increasing microwave power and time. However, cooling time during irradiation did not affect the extraction efficiency. Optimal conditions were identified at pH of 1, ratio of 2:100 g/mL, 6 min irradiation, and microwave power of 960 W. Under these optimal conditions, approximately 50.55 mg phenolics could be recovered from 1 g dried peel. These conditions are recommended for recovery of phenolic compounds from banana peel for further utilisation.

Banana peel, a by-product rich in phenolics and other bioactive compounds, has great potentials as a natural preservative or healthy food ingredient. However, the instability of bioactive compounds derived from banana peel limits their applications, and as such encapsulation is necessary to improve their stability and widen their applications. This study investigated the impact of spray drying conditions and coating materials on the physical, phytochemical, and antioxidant properties of the peel extract to identify the most suitable encapsulation process. The results showed that inlet temperature (ranging from 140 to 180 °C) and feeding rate (3–15 mL/min) did not significantly affect the total phenolic content (TPC) and antioxidant capacity but influenced the moisture content and recovery yield of the powder. The ratio of dry matter in fresh extract-to-coating material (DM-to-CM) (1:1–1:7 (w/w)) did not affect the moisture content. However, it affected the TPC, antioxidant properties, and recovery yield of the powder. Finally, the type of coating materials did not significantly affect TPC and antioxidant properties, but other physical properties, dopamine levels and recovery yield. The most suitable encapsulation conditions were identified as an inlet drying temperature of 150 °C, a feeding rate of 9 mL/min, a ratio of DM-to-CM of 1:1 (w/w), and coating with a combination of maltodextrin M100 and gum acacia. Powder prepared under the most suitable conditions had a spherical shape with a rough surface and had stable TPC under storage conditions of 40 °C for 4 weeks. It also has ideal physical, phytochemical and antioxidant properties and is suitable for further applications.

To determine the diagnostic and prognostic significance of growth factors and the urokinase-type plasminogen activator (uPA) system in pancreatic ductal adenocarcinoma (PDAC) using a multigene assay.Messenger RNA (mRNA) expression of 15 genes from epidermal growth factor receptor, insulin-like growth factor (IGF), and uPA families were measured in 46 PDAC tissue samples using quantitative real-time reverse transcription-polymerase chain reaction. These results were compared with those of the uninvolved adjacent (AP) tissue and benign mucinous cystadenomas (BMC). The mRNA expression was evaluated using logistic regression and receiver operating characteristic area under the curve (ROC AUC) analyses. Their relationship with prognosis was tested by Cox regression multivariate analysis.All genes were overexpressed in most of the PDAC tissue. When compared with AP tissue, the median expression values for IGF-binding protein 3 (IGFBP-3) and uPA receptor (uPAR) was 9.8- and 9.6-fold, respectively. Expression levels of uPA, uPAR, IGF-I, and IGFBP-3 mRNA were significantly greater in PDAC than in BMC. The IGFBP-3 mRNA expression demonstrated greatest ROC AUC values for PDAC versus AP tissue (ROC AUC, 0.745; 95% confidence interval [CI], 0.65-0.86); whereas ROC AUC values were greatest for uPAR when PDAC was compared with BMC (ROC AUC, 0.846; 95% CI, 0.76-0.94). The combination of uPA, uPAR, and IGF-I significantly improved discriminatory power (ROC AUC, 0.965; 95% CI, 0.93-1.00). The IGFBP-3, uPA, plasminogen activator inhibitor-2, and International Union Against Cancer stage had a significant influence on survival, but the effect of IGFBP-3 was lost after multivariate stepwise analysis.These results indicate that there is an influence of IGF system in tumor progression from BMC to PDAC, whereas the uPA/uPAR system has the greater influence on survival in PDAC.

The serum/plasma proteome was explored for biomarkers to improve the diagnostic ability of CA19-9 in pancreatic adenocarcinoma (PC).A Training Set of serum samples from 20 resectable and 18 stage IV PC patients, 54 disease controls (DCs) and 68 healthy volunteers (HVs) were analysed by surface-enhanced laser desorption and ionisation time-of-flight mass spectrometry (SELDI-TOF MS). The resulting protein panel was validated on 40 resectable PC, 21 DC and 19 HV plasma samples (Validation-1 Set) and further by ELISA on 33 resectable PC, 28 DC and 18 HV serum samples (Validation-2 Set). Diagnostic panels were derived using binary logistic regression incorporating internal cross-validation followed by receiver operating characteristic (ROC) analysis.A seven-protein panel from the training set PC vs DC and from PC vs HV samples gave the ROC area under the curve (AUC) of 0.90 and 0.90 compared with 0.87 and 0.91 for CA19-9. The AUC was greater (0.97 and 0.99, P<0.05) when CA19-9 was added to the panels and confirmed on the validation-1 samples. A simplified panel of apolipoprotein C-I (ApoC-I), apolipoprotein A-II (ApoA-II) and CA19-9 was tested on the validation-2 set by ELISA, in which the ROC AUC was greater than that of CA19-9 alone for PC vs DC (0.90 vs 0.84) and for PC vs HV (0.96 vs 0.90).A simplified diagnostic panel of CA19-9, ApoC-I and ApoA-II improves the diagnostic ability of CA19-9 alone and may have clinical utility.

Xao tam phan (Paramignya trimera (Oliv.) Guillaum) has been used as an herbal medicine for the treatment of cancer or cancer-like diseases in recent years, particularly in Vietnam. Drying is an important step for preparation of dried materials for storage and further investigation; however, the effects of drying must be taken into account when processing samples, because this can have profound effects on the stability of phytochemical compounds and the biological activity of the dried P. trimera root. As such, this study assessed the effects of four different drying methods (conventional, hot air, vacuum, and microwave) on phytochemical retention and antioxidant capacity of P. trimera root, to identify an optimal drying method for P. trimera root. The results showed that the drying methods significantly affected phytochemical levels and antioxidant capacity of P. trimera root and that among the four drying methods tested, microwave drying (400 W) had the highest levels of phytochemical compounds, with total phenolic, total flavonoid, proanthocyanidin, and saponin contents of 11.27 mg GAE, 19.88 mg RE, 3.98 mg CE, and 267.15 mg EE/gram of dried sample, respectively. Dried sample prepared using this method had antioxidant capacity comparable to that of other drying methods. In addition, this method had the shortest drying time (0.28 h) and consumed the least energy (0.28 kWh). Therefore, microwave drying should be considered for drying P. trimera root for further investigation and utilization.

Ovarian cancer is the most common cause of death among women with gynecologic cancer. We examined molecular profiles of fibroblasts from normal ovary and high-grade serous ovarian tumors to identify novel therapeutic targets involved in tumor progression. We identified 2,300 genes that are significantly differentially expressed in tumor-associated fibroblasts. Fibroblast expression of one of these genes, connective tissue growth factor (CTGF), was confirmed by immunohistochemistry. CTGF protein expression in ovarian tumor fibroblasts significantly correlated with gene expression levels. CTGF is a secreted component of the tumor microenvironment and is being pursued as a therapeutic target in pancreatic cancer. We examined its effect in in vitro and ex vivo ovarian cancer models, and examined associations between CTGF expression and clinico-pathologic characteristics in patients. CTGF promotes migration and peritoneal adhesion of ovarian cancer cells. These effects are abrogated by FG-3019, a human monoclonal antibody against CTGF, currently under clinical investigation as a therapeutic agent. Immunohistochemical analyses of high-grade serous ovarian tumors reveal that the highest level of tumor stromal CTGF expression was correlated with the poorest prognosis. Our findings identify CTGF as a promoter of peritoneal adhesion, likely to mediate metastasis, and a potential therapeutic target in high-grade serous ovarian cancer. These results warrant further studies into the therapeutic efficacy of FG-3019 in high-grade serous ovarian cancer.

Australia is home to over 800 different species of Eucalyptus and traditionally, many Eucalyptus species have been utilised to heal wounds and treat fungal infections by the Indigenous people of Australia. In view of this, our study was designed to investigate the phytochemical, antibacterial and antifungal properties of crude aqueous extract of E. microcorys leaves. The freeze-dried powdered extract was prepared and the phytochemical profile was studied by analysing the total phenolic content (TPC), total flavonoid content (TFC), proanthocyanidins, antioxidants and saponins. The TPC, TFC and proanthocyanidin values found were: 501.76 ± 14.47 mg of gallic acid equivalents per g, 61.53 ± 0.83 mg of rutin equivalents per g and 10.76 ± 0.89 mg of catechin equivalents per g, respectively. The antioxidant values expressed in mg trolox equivalents per g of extract (mg TE/g) were: ABTS = 1073.13 ± 10.73 mg TE/g, DPPH = 1035.44 ± 65.54 mg TE/g and CUPRAC = 1524.30 ± 66.43 mg TE/g. The powdered extract was also evaluated for activity against three pathogenic bacterial strains (Escherichia coli, Enterobacter aerogenes, Staphylococcus lugdunensis); and three fungal strains (Geotrichum candidum, Aspergillus brasiliensis and Candida albicans) using the disc diffusion method and 96 well plate-based method with resazurin dye. The extract exhibited clear zones of inhibition against the tested bacteria and fungi. Minimum inhibitory concentration (MIC) values were demonstrated to be: A. brasiliensis = 2.44 μg/mL, G. candidum = 4.88 μg/mL, S. lugdunensis = 78 μg/mL, E. coli = 156.25 μg/mL, E. aerogenes = 312.5 μg/mL and C. albicans = 1250 μg/mL. These results reveal the significant potential of E. microcorys as a source of phenolics, antioxidants and antimicrobial agents and also highlight the necessity of further purification and characterisation of solitary bioactive compounds for their prospective applications in food, nutraceutical and pharmaceutical industries.

Lilly pilly (LP) fruit (Syzygium paniculatum Gaertn.) is widely grown in eastern Australia and has been used as food by indigenous Australians. However, there is limited information on its bioactivity. This study investigated the physicochemical and antioxidant properties of the crude fruit extract, identified its bioactive compounds and also assessed its potential anti-proliferative effect on pancreatic cancer cells. Our data showed that the LP extract was water-soluble and possessed a total phenolic content of 96 mg of gallic acid equivalents (GAE)/g, flavonoid levels of 52 mg catechin equivalents (CAE)/g, proanthocyanidin levels of 29 mg CAE/g. Several phenolic compounds such as gallic acid, chlorogenic acid, catechin and epicatechin were identified in the LP extract with levels of 0.39, 2.35, 0.47 and 2.9 mg/g, respectively. Results from six different antioxidant assays revealed that the LP extract pocessed potent antioxidant and free radical scavenging capacity. Although antioxidant capacity of the extract was lower than that of vitamin E, vitamin C and BHT, it could be significantly improved if the extract was to be further purified. We also showed that the LP extract (200 μg/mL) significantly reduced the viability of MiaPaCa-2 and ASPC-1 pancreatic cancer cells to levels comparable to that of the chemotherapeutic agent gemcitabine. For this reason lilly pilly should be further investigated for its health promoting and potential anti-cancer benefits, particularly for pancreatic cancer.

The effect of microwave pretreatment on the levels of total phenolic compounds, flavonoids, proanthocyanidins, and individual major compounds as well as the total antioxidant activity of the dried lemon pomace was investigated. The results showed that microwave pretreatment significantly affected all the examined parameters. The total phenolic content, total flavonoids, proanthocyanidins, as well as the total antioxidant activity significantly increased as the microwave radiation time and power increased (e.g., 2.5-fold for phenolics, 1.4-fold for flavonoids, and 5.5-fold for proanthocyanidins); however, irradiation of more than 480 W for 5 min resulted in the decrease of these parameters. These findings indicate that microwave irradiation time and power may enhance higher levels of the phenolic compounds as well as the antioxidant capacity of the dried lemon pomace powder. However, higher and longer irradiation may lead to a degradation of phenolic compounds and lower the antioxidant capacity of the dried lemon pomace. Practical applications Lemon pomace could be a good source of bioactive compounds and antioxidants. Microwave irradiation could be applied for the enhancement of the total phenolic compounds and antioxidants of the lemon pomace-dried powder. The findings of this study can be applied for enhancing the bioactive compounds and the antioxidant activity of the dried lemon pomace for further extraction, isolation, and utilisation.

Journal of Food Processing and PreservationVolume 41, Issue 1 e12879 Original Article Bioactive Compound Yield and Antioxidant Capacity of Helicteres hirsuta Lour. Stem as Affected by Various Solvents and Drying Methods Hong Ngoc Thuy Pham, Corresponding Author Hong Ngoc Thuy Pham hongngocthuy.pham@uon.edu.au orcid.org/0000-0002-6722-8910 School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 Australia Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu street, Nha Trang City, Khanh Hoa, 8458 VietnamCorresponding author. TEL: + 61 243484680 or + 61 434085481; FAX: + 61 243484145; EMAIL: c.scarlett@newcastle.edu.au or hongngocthuy.pham@uon.edu.auSearch for more papers by this authorVan Tang Nguyen, Van Tang Nguyen School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 Australia Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu street, Nha Trang City, Khanh Hoa, 8458 VietnamSearch for more papers by this authorQuan Van Vuong, Quan Van Vuong School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 AustraliaSearch for more papers by this authorMichael C. Bowyer, Michael C. Bowyer School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 AustraliaSearch for more papers by this authorChristopher J. Scarlett, Corresponding Author Christopher J. Scarlett c.scarlett@newcastle.edu.au School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 AustraliaCorresponding author. TEL: + 61 243484680 or + 61 434085481; FAX: + 61 243484145; EMAIL: c.scarlett@newcastle.edu.au or hongngocthuy.pham@uon.edu.auSearch for more papers by this author Hong Ngoc Thuy Pham, Corresponding Author Hong Ngoc Thuy Pham hongngocthuy.pham@uon.edu.au orcid.org/0000-0002-6722-8910 School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 Australia Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu street, Nha Trang City, Khanh Hoa, 8458 VietnamCorresponding author. TEL: + 61 243484680 or + 61 434085481; FAX: + 61 243484145; EMAIL: c.scarlett@newcastle.edu.au or hongngocthuy.pham@uon.edu.auSearch for more papers by this authorVan Tang Nguyen, Van Tang Nguyen School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 Australia Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu street, Nha Trang City, Khanh Hoa, 8458 VietnamSearch for more papers by this authorQuan Van Vuong, Quan Van Vuong School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 AustraliaSearch for more papers by this authorMichael C. Bowyer, Michael C. Bowyer School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 AustraliaSearch for more papers by this authorChristopher J. Scarlett, Corresponding Author Christopher J. Scarlett c.scarlett@newcastle.edu.au School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, NSW, 2258 AustraliaCorresponding author. TEL: + 61 243484680 or + 61 434085481; FAX: + 61 243484145; EMAIL: c.scarlett@newcastle.edu.au or hongngocthuy.pham@uon.edu.auSearch for more papers by this author First published: 17 June 2016 https://doi.org/10.1111/jfpp.12879Citations: 31Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Abstract Different parts of Helicteres hirsuta Lour. (H. hirsuta L.) have been used as traditional medicines, however limited studies have been conducted on the preparation of dried material or the extraction of bioactive compounds. This study aimed to determine the effect of various solvents and drying methods on the bioactive compound yield and antioxidant capacity of the H. hirsuta L. stem. The results showed that water was the best solvent for obtaining the highest levels of phenolics and flavonoids, whereas methanol was the solvent of choice for saponin extraction. Among assessed drying methods, hot-air drying at 80C revealed as the best method to maintain the bioactive components (phenolics of 8.99 mg GAE/g, flavonoids of 9.60 mg CE/g and saponins of 15.18 mg ESE/g), and DPPH, ABTS, and FRAP antioxidant activities. Thus hot-air drying at 80C is recommended for preparation of dried H. hirsuta L. stem for further processing steps. Practical Applications The optimal drying condition to prepare dried H. hirsuta L. stem is hot-air drying at 80C, a simple and fast drying method, and the most suitable solvent for extraction of antioxidant components from dried H. hirsuta L. stem is water; a cheap, accessible, and environmentally friendly solvent. These conditions can be applied for isolating and purifying bioactive compounds for further application in the food and pharmaceutical industries. Citing Literature Volume41, Issue1February 2017e12879 RelatedInformation

Edible films and coatings have been applied as the potential substitutes for conventional plastics in food packaging. However, their physical and mechanical properties still have limitations and thus require further improvement. In this study, we compared the physico–chemical properties of starches extracted from eight rice varieties and attempted to predict their promising effects on the physical (thickness and solubility), mechanical (tensile strength and elongation break), barrier (water vapor permeability), and optical properties (color and transparency) of rice starch‐ι‐carrageenan films. The results showed that starch amylose content and amylose–amylopectin associations during retrogradation play a significant role in determining various properties of the films. The film containing starch from “Reiziq” variety showed minimum thickness (0.08 mm), water vapor permeability (WVP) (2.7 gs −1 m −1 Pa −1 ), solubility (43.12%) opacity (0.44%), and better mechanical properties, demonstrating the importance of selection of the source of the starch. The results also indicated that rice starch had compatibility with ι‐carrageenan, and the blend of these two polysaccharides can be potentially used for coating fruit and vegetables.

This study aimed to optimise microwave-assisted extraction (MAE) conditions for total phenolic compounds (TPCs) and antioxidant activities of the alga Sargassum vestitum by using response surface methodology with Box–Behnken design. The results showed that solvent concentration had the greatest impact on TPC and antioxidant activities of the extracts, followed by radiation time and power. The optimal MAE conditions were ethanol concentration of 70%, radiation time of 75 s and power of 80%. The optimal MAE method showed much better extraction efficacy of phenolics and antioxidant capacities of the extract than conventional and ultrasonic methods.

This study demonstrated Response Surface Methodology (RSM) optimisation of starch edible coating formulation and its application on apple fruit quality and storage life at two different temperatures (20 °C and 5 °C + 1 day at 20 °C). Films were optimised for physical, mechanical and permeability properties and the optimal formulation was sprayed over the fruit surface. Fruit weight loss, respiration rate, total soluble solids (TSS), titratable acidity (TA), firmness, greasiness and change in the fruit skin colour were measured. The effect of the coating treatment on changes to the bioactive profile (phenolics and free radical scavenging activity) of the fruit was also analysed. The results obtained from this study showed that the optimised formulation (rice starch 2.5% ι-car 1.5% sucrose fatty acid ester 2% glycerol 1.5%) in combination with low temperature was effective in reducing weight loss and maintained tissue firmness without affecting TSS, TA and the bioactive profile of the apple fruit during postharvest storage. A delay in skin colour change and a significant reduction in fruit greasiness was observed in coated fruit (p < 0.05). These results show the potential of starch-based coating formulation in the formulation in improving the visual appearance, without affecting the fruit internal quality and nutritional properties of apple fruit during storage.

Intense sweeteners (IS) are often marketed as a healthier alternative to sugars, with the potential to aid in combating the worldwide rise of diabetes and obesity. However, their use has been counterintuitively associated with impaired glucose homeostasis, weight gain and altered gut microbiota. The nature of these associations, and the mechanisms responsible, are yet to be fully elucidated. Differences in their interaction with taste receptors may be a potential explanatory factor. Like sugars, IS stimulate sweet taste receptors, but due to their diverse structures, some are also able to stimulate bitter taste receptors. These receptors are expressed in the oral cavity and extra-orally, including throughout the gastrointestinal tract. They are involved in the modulation of appetite, glucose homeostasis and gut motility. Therefore, taste genotypes resulting in functional receptor changes and altered receptor expression levels may be associated with metabolic conditions. IS and taste receptors may both interact with the gastrointestinal microbiome, and their interactions may potentially explain the relationship between IS use, obesity and metabolic outcomes. While these elements are often studied in isolation, the potential interactions remain unexplored. Here, the current evidence of the relationship between IS use, obesity and metabolic outcomes is presented, and the potential roles for interactions with taste receptors and the gastrointestinal microbiota in modulating these relationships are explored.

Catharanthus roseus (C. roseus) is an important medicinal plant distributed in many countries. It has attracted increasing attention due to it being shown to possess a range of phytochemicals with various biological activities such as antioxidant, antibacterial, antifungal, antidiabetic and anticancer properties. Remarkably, vinblastine and vincristine isolated from this plant were the first plant-derived anticancer agents deployed for clinical use. Recently, new isolated indole alkaloids from this plant including catharoseumine, 14′,15′-didehydrocyclovinblastine, 17-deacetoxycyclovinblastine and 17-deacetoxyvinamidine effectively inhibited human cancer cell lines in vitro. Moreover, vindoline, vindolidine, vindolicine and vindolinine isolated from C. roseus leaf exhibited in vitro antidiabetic property. These findings strongly indicate that this plant is still a promising source of bioactive compounds, which should be further investigated. This paper provides an overview of the traditional use and phytochemical profiles of C. roseus, and summarises updated techniques of the preparation of dried material, extraction and isolation of bioactive compounds from this plant. In addition, purported health benefits of the extracts and bioactive compounds derived from this plant were also addressed to support their potential as therapeutic agents.

HER-2 is a transmembrane growth factor receptor recognized in overexpression as an independent adverse prognostic factor in several cancers. This study measured HER-2 overexpression in pancreatic adenocarcinoma at the genetic, transcriptional, and translational level. Expression was gauged with regard to stage, grade, and survival. Pancreatic adenocarcinoma samples (n = 30) were analyzed with immunohistochemical labeling for HER-2 protein, Quantitative real-time reverse transcriptase polymerase chain reaction (Q-RT-PCR) measurement of HER-2 mRNA and fluorescence in situ hybridization (FISH) analysis of HER-2 gene expression. HER-2 expression in benign pancreatic lesions (n = 10) provided a control. Five (17%) of the pancreatic adenocarcinomas scored maximal 3+ immunohistochemistry (IHC) labeling, seven (23%) had significantly increased expression of HER-2 mRNA, while only one (3%) exhibited low level HER-2 gene amplification. Ten (33%) tumors demonstrated aneuploidy. In general, concordance between methodologies was poor, but the best agreement was seen between FISH aneuploidy status and Q-RT-PCR mRNA overexpression (80% agreement), followed by IHC and Q-RT-PCR (73% agreement). The least agreement was seen between IHC and FISH aneuploidy status (67% agreement). Tumor stage was positively associated with HER-2 mRNA and protein expression, but tumor grade and other patient characteristics did not reach statistical significance. A poor survival outcome was demonstrated with positive HER-2 status in all three measures of overexpression (Kaplan-Meier log-rank score; P < 0.01 [IHC], P = 0.05 [Q-RT-PCR], P = 0.02 [FISH]). Discordance in expression at the nuclear, cytoplasmic, and cell surface levels highlights the limitations of immunohistochemical evaluation alone and stresses the need for further evaluation of response to anti-HER-2 targeted therapies in tumors displaying overexpression in gene copy, mRNA, and receptor protein.

The N-Myc oncoprotein induces neuroblastoma, which arises from undifferentiated neuroblasts in the sympathetic nervous system, by modulating gene and protein expression and consequently causing cell differentiation block and cell proliferation. The class IIa histone deacetylase 5 (HDAC5) represses gene transcription, and blocks myoblast, osteoblast and leukemia cell differentiation. Here we showed that N-Myc upregulated HDAC5 expression in neuroblastoma cells. Conversely, HDAC5 repressed the ubiquitin–protein ligase NEDD4 gene expression, increased Aurora A gene expression and consequently upregulated N-Myc protein expression. Genome-wide gene expression analysis and protein co-immunoprecipitation assays revealed that HDAC5 and N-Myc repressed the expression of a common subset of genes by forming a protein complex, whereas HDAC5 and the class III HDAC SIRT2 independently repressed the expression of another common subset of genes without forming a protein complex. Moreover, HDAC5 blocked differentiation and induced proliferation in neuroblastoma cells. Taken together, our data identify HDAC5 as a novel co-factor in N-Myc oncogenesis, and provide the evidence for the potential application of HDAC5 inhibitors in the therapy of N-Myc-induced neuroblastoma and potentially other c-Myc-induced malignancies.

Euphorbia tirucalli (E. tirucalli) is now widely distributed around the world and is well known as a source of traditional medicine in many countries. This study aimed to utilise response surface methodology (RSM) to optimise ultrasonic-assisted extraction (UAE) conditions for total phenolic compounds (TPC) and antioxidant capacity from E. tirucalli leaf. The results showed that ultrasonic temperature, time and power effected TPC and antioxidant capacity; however, the effects varied. Ultrasonic power had the strongest influence on TPC; whereas ultrasonic temperature had the greatest impact on antioxidant capacity. Ultrasonic time had the least impact on both TPC and antioxidant capacity. The optimum UAE conditions were determined to be 50 °C, 90 min. and 200 W. Under these conditions, the E. tirucalli leaf extract yielded 2.93 mg GAE/g FW of TPC and exhibited potent antioxidant capacity. These conditions can be utilised for further isolation and purification of phenolic compounds from E. tirucalli leaf.

Eucalyptus robusta (Sm.) (ER) is a widely distributed tree native to the east coast of Australia, which has also been established in numerous other countries. ER leaves contain high levels of essential oils and are rich in total phenolic compounds (TPC), which have been linked with health benefits; however, there is limited information on the bioactivity of ER leaf extracts. This study aimed to optimise water extraction conditions for TPC, prepare a spray-dried powdered extract and test its physicochemical, antioxidant and anti-proliferative properties. The results showed that optimal water extraction conditions for TPC were 85 °C, 15 min and a water-to-leaf ratio of 20:1 mL/g. Under these conditions, spray-dried powdered extract was prepared with a recovery yield of 85%. The extract was water-soluble and had a TPC level of 407 mg GAE/g. It also possessed potent antioxidant capacity, comparable to pure ascorbic acid, but higher than pure α-tocopherol. In addition, the powdered extract demonstrated significant activity against a panel of cancer cell lines, which included cancers of the pancreas, breast, lung, brain, skin, colon and ovary. Of note, the ER extract exerted a more significant toxic effect on pancreatic cancer (PC) cells compared to gemcitabine, the first line chemotherapeutic agent for PC. We suggest that future studies should purify individual bioactive compounds from ER for further investigation of its potential health promoting and anti-cancer activity.

Summary The aim of this study was to optimise the aqueous extraction conditions for the recovery of phenolic compounds and antioxidant capacity of lemon pomace using response surface methodology. An experiment based on Box–Behnken design was conducted to analyse the effects of temperature, time and sample‐to‐water ratio on the extraction of total phenolic compounds, total flavonoids, proanthocyanidins and antioxidant capacity. Sample‐to‐solvent ratio had a negative effect on all the dependent variables, while extraction temperature and time had a positive effect only on TPC yields and ABTS antioxidant capacity. The optimal extraction conditions were 95 °C, 15 min and a sample‐to‐solvent ratio of 1:100 g mL −1 . Under these conditions, the aqueous extracts had the same content of TPC and TF as well as antioxidant capacity in comparison with those of methanol extracts obtained by sonication. Therefore, these conditions could be applied for further extraction and isolation of phenolic compounds from lemon pomace.

A large amount of citrus waste is generated annually. This waste is of great economic worth, since it contains high levels of polyphenols, which have attracted scientific interest due to their potent antimicrobial and antiradical activities. Pretreatment is a crucial step that precedes the extraction process and influences the yields and quality of polyphenols. This review emphasizes the effect of different drying processes, such as freeze drying, hot-air drying, vacuum drying, microwave drying, infrared drying, and high-speed drying, on the polyphenol retention in citrus by-products. Further treatments of the dried citrus by-products for assisting the liberation of bound polyphenols are also provided and comprehensively discussed.

The moisture sorption isotherm of pea starch films prepared with various glycerol contents as plasticizer was investigated at different storage relative humidities (11%–96% RH) and at 5 ± 1, 15 ± 1, 25 ± 1 and 40 ± 1 °C by using gravimetric method. The results showed that the equilibrium moisture content of all films increased substantially above aw = 0.6. Films plasticized with glycerol, under all temperatures and RH conditions (11%–96%), adsorbed more moisture resulting in higher equilibrium moisture contents. Reduction of the temperature enhanced the equilibrium moisture content and monolayer water of the films. The obtained experimental data were fitted to different models including two-parameter equations (Oswin, Henderson, Brunauer–Emmitt–Teller (BET), Flory–Huggins, and Iglesias–Chirife), three-parameter equations Guggenhiem–Anderson–deBoer (GAB), Ferro–Fontan, and Lewicki) and a four-parameter equation (Peleg). The three-parameter Lewicki model was found to be the best-fitted model for representing the experimental data within the studied temperatures and whole range of relative humidities (11%–98%). Addition of glycerol increased the net isosteric heat of moisture sorption of pea starch film. The results provide important information with estimating of stability and functional characteristics of the films in various environments.

Journal of Food Processing and PreservationVolume 41, Issue 2 e12851 Original Article Microwave-Assisted Extraction for Saponins and Antioxidant Capacity from Xao Tam Phan (Paramignya trimera) Root Van Tang Nguyen, Corresponding Author Van Tang Nguyen vantang.nguyen@uon.edu.au School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, New South Wales, 2258 Australia Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamCorresponding authors. TEL: +61 434238842; FAX: +61 243484145; EMAIL: vantang.nguyen@uon.edu.au or c.scarlett@newcastle.edu.auSearch for more papers by this authorQuan V. Vuong, Quan V. Vuong Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamSearch for more papers by this authorMichael C. Bowyer, Michael C. Bowyer Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamSearch for more papers by this authorIan A. Van Altena, Ian A. Van Altena Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamSearch for more papers by this authorChristopher J. Scarlett, Corresponding Author Christopher J. Scarlett c.scarlett@newcastle.edu.au Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamCorresponding authors. TEL: +61 434238842; FAX: +61 243484145; EMAIL: vantang.nguyen@uon.edu.au or c.scarlett@newcastle.edu.auSearch for more papers by this author Van Tang Nguyen, Corresponding Author Van Tang Nguyen vantang.nguyen@uon.edu.au School of Environmental and Life Sciences, Faculty of Science and Information Technology, University of Newcastle, Ourimbah, New South Wales, 2258 Australia Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamCorresponding authors. TEL: +61 434238842; FAX: +61 243484145; EMAIL: vantang.nguyen@uon.edu.au or c.scarlett@newcastle.edu.auSearch for more papers by this authorQuan V. Vuong, Quan V. Vuong Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamSearch for more papers by this authorMichael C. Bowyer, Michael C. Bowyer Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamSearch for more papers by this authorIan A. Van Altena, Ian A. Van Altena Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamSearch for more papers by this authorChristopher J. Scarlett, Corresponding Author Christopher J. Scarlett c.scarlett@newcastle.edu.au Department of Food Technology, Faculty of Food Technology, Nha Trang University, No. 2 Nguyen Dinh Chieu, Nha Trang, Khanh Hoa, 8458 VietnamCorresponding authors. TEL: +61 434238842; FAX: +61 243484145; EMAIL: vantang.nguyen@uon.edu.au or c.scarlett@newcastle.edu.auSearch for more papers by this author First published: 15 June 2016 https://doi.org/10.1111/jfpp.12851Citations: 23Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Abstract The aim of this study was to determine the optimal microwave-assisted extraction (MAE) parameters for saponins and antioxidant capacity from the P. trimera root. The optimal MAE parameters were found to be 100% methanol, extraction time of 40 min and ratio of solvent to sample of 100 mL/g. Saponin content, saponin extraction efficiency, ABTS radical scavenging capacity, DPPH radical scavenging capacity, cupric ion reducing antioxidant capacity, and ferric reducing antioxidant power of the P. trimera root achieved under these parameters were 520.5 mg escin equivalents (EE)/g dried sample, 62.6%, 215.3, 93.2, 24.7, 121.7 mg trolox equivalents (TE)/g dried sample, respectively, which were higher than predicted values (488.8 mg EE/g dried sample, 58.8%, 213.7, 89.5, 24.0, 115.8 mg TE/g dried sample, respectively). Therefore, the optimal MAE parameters of 100% methanol, 40 min and 100 mL/g are recommended for obtaining saponin-enriched extracts from the P. trimera root for potential application in the food industry. Practical Application This study indicated that methanol concentration had the highest effect on the extractability of saponins and antioxidant capacity from P. trimera root, followed by ratio of solvent to sample and extraction time. Therefore, the application of the optimized MAE parameters allows obtaining the highest saponin yield as well as the greatest antioxidant capacity from P. trimera root for further investigation and application. Citing Literature Volume41, Issue2April 2017e12851 RelatedInformation

The Davidson's plum (DP; Davidsonia pruriens F. Muell) is a fruit native to Australia that is a rich source of phenolic compounds. This study investigated the effects of various extraction solvents (ethanol, methanol, acetone and water) on total phenolic content (TPC), antioxidant capacity (ABTS, DPPH, CUPRAC, FRAP assays) and anti-proliferative activity (MTT, CCK-8 assays) of DP. Our data revealed that extraction solvents significantly affected the physico-chemical, antioxidant and anti-proliferative properties of DP extracts. Within the tested solvents, ethanol was found to be the optimal solvent for extraction of bioactive compounds from DP as its extract had the greatest TPC (94.13 GAE/g), flavonoids (78.33 mg RUE/g), proanthocyanidins (5.33 mg CAE/g), anthocyanidins (2.81 mg CGE/g), as well as possessed the highest antioxidant capacity and anti-proliferative activity against a panel of cancer cell lines. This included cancers of the pancreas, breast, lung, brain, skin, colon and ovary. Therefore, further investigations should be conducted to identify key bioactive compounds to determine the potential for utilization in the nutraceutical and phytopharmaceutical industries.

The type 2 family of taste receptors (T2Rs) detect and respond to bitter tastants. These receptors are expressed throughout the gastrointestinal (GI) tract, with location dependant roles. In the oral cavity, T2Rs are involved in the conscious perception of bitter tastants, while in the lower GI tract they have roles in chemoreception and regulation of GI function. Through these diverse roles, these receptors may be involved in modulating appetite and diet, with consequences for weight regulation and obesity. Interestingly, the concentration of T2Rs in the GI tract is greatest in the large intestine, the organ with the densest colonisation of bacteria. The gut microbiome has been the subject of intense research, as a plethora of roles linking microbiota to human health continue to be uncovered. Of particular interest is the microbial signature associated with obesity. Obesity is a leading health concern, and advances in our understanding of this disease are needed. Diet is a known modifiable factor in the development of obesity. However, diet only partially explains disease risk. Changes in microbial energy harvesting by the microbiota plays a role in obesity, and the composition of these energy harvesting populations may be controlled by taste receptors. This review explores T2Rs as a potential link between obesity and the human GI microbiome.

The individual and interactive impacts of guar gum and glycerol on the pea‐starch‐based edible film characteristics were examined using three factors with three‐level Box–Behnken response surface design (BBD). The results showed that density and elongation at break were only significantly ( p &lt; 0.05) affected by pea starch and guar gum in a positive linear fashion. The quadratic regression coefficient of pea starch showed a significant effect ( p &lt; 0.05) on thickness, density, puncture force, water vapor permeability, and tensile strength, while tensile strength and Young's modulus were affected by the quadratic regression coefficient of glycerol and guar gum, respectively. The results were analyzed using Pareto analysis of variance (ANOVA) and the developed predictive equations for each response variable presented reliable and satisfactory fit with high coefficient of determination ( R 2 ) values (≥0.96). The optimized conditions with the goal of maximizing mechanical properties and minimizing water vapor permeability were 2.5 g pea starch, 0.3 g guar gum, and 25% w/w glycerol based on the dry film matter in 100 mL of distilled water.

Several studies have shown that UV-C (ultraviolet C) irradiation promotes the bioactive compounds and antioxidants of fresh fruits and vegetables. The aim of this study was to apply UV irradiation in dried lemon pomace powder for enhancing its phenolic content and antioxidant properties, thus more bioactive compounds should be available for extraction and utilization. Lemon pomace dried powder was placed under a UV lamp and treated with dosages of 4, 19, 80 and 185 kJ·m-2, while untreated powder was used as a control. UV-C irradiation significantly affected the total phenolic content, total flavonoid content, proanthocyanidins, and antioxidant capacity measured by cupric reducing antioxidant capacity (CUPRAC) and ferric reducing antioxidant power (FRAP) of the lemon pomace dried powder, while it did not affect the vitamin C content. UV-C irradiation of 19 kJ·m-2 resulted in 19% higher total phenolic content than the control, while UV-C irradiation of 180 kJ·m-2 resulted in 28% higher total flavonoid content than the control. The antioxidant capacity was reduced when UV-C irradiation more than 4 kJ·m-2 was applied. The results of this study indicate that UV-C treatment has the potential to increase the extraction of bioactive compounds of dried lemon pomace at relatively high dosages.

The aims of this study were to screen the phytochemical content, antioxidant, antimicrobial and cytotoxic properties of the extract and its fractions prepared from Catharanthus roseus (L.) G. Don (C. roseus) stem. C. roseus stem was powdered and extracted with methanol using ultrasound-assisted extraction to obtain the crude extract. The crude extract was further fractioned using liquid-liquid extraction technique to obtain extracts of increasing polarity including n-butanol and residual aqueous fractions. The crude extract and its derived fractions were then subjected to phytochemical screening and assayed for antioxidant, antimicrobial and cytotoxic activities. Results showed that the n-butanol fraction contained the highest levels of saponins and phenolics (3037.54 mg ESE/g and 77.87 mg GAE/g, respectively) and possessed the strongest antioxidant capacity amongst the tested extracts. HPLC analysis revealed that this n-butanol fraction had high levels of apigenin and kaempferol, whereas the aqueous fraction contained a high level of gallic acid. The n-butanol fraction was found to effectively inhibit the activity of Escherichia coli and Staphylococccus lugdunensis. The n-butanol fraction also possessed strong cytotoxic activity in vitro against a wide range of cancer cell lines including A2780 (ovarian), H460 (lung), A431 (skin), MIA PaCa-2 (pancreas), Du145 (prostate), HT29 (colon), MCF-7 (breast), BE2-C (neuroblastoma), SJ-G2, U87 and SMA (glioblastoma) at low doses (GI50 values of 5.2−21.0 µg/mL). These results indicate that the n-butanol fraction prepared from C. roseus stem is a rich source of bioactive compounds which can be isolated for further evaluation as potential antimicrobial drugs or antitumor therapeutic agents.

Optimal folate levels may prevent endothelial dysfunction in inflammatory diseases.• Folate may also alter inflammatory responses via DNA methylation and synthesis processes.• The link between folate and inflammation varies based on several factors, such as timing of intervention.• Further studies are needed before making folate intake recommendations around inflammation.

Pancreatic cancer (PC), with a 5 year survival of <7%, is one of the most fatal of all human cancers. The highly aggressive and metastatic character of this disease poses a challenge that current therapies are failing, despite significant efforts, to meet. This review examines the current status of the 35 small molecule inhibitors targeting pancreatic cancer in clinical trials and the >50 currently under investigation. These compounds inhibit biological targets spanning protein kinases, STAT3, BET, HDACs and Bcl-2 family proteins. Unsurprisingly, protein kinase inhibitors are overrepresented. Some trials show promise; a phase I combination trial of vorinostat 11 and capecitabine 17 gave a median overall survival (MoS) of 13 months and a phase II study of pazopanib 15 showed a MoS of 25 months. The current standard of care for metastatic pancreatic ductal adenocarcinoma, fluorouracil/folic acid (5-FU, Adrucil®), and gemcitabine (GEMZAR®) afforded a MoS of 23 and 23.6 months (EPAC-3 study), respectively. In patients who can tolerate the FOLFIRINOX regime, this is becoming the standard of treatment with a MoS of 11.1 months. Clinical study progress has been slow with limited improvement in patient survival relative to gemcitabine 1 monotherapy. A major cause of low PC survival is the late stage of diagnosis, occurring in patients who consider typical early stage warning signs of aches and pains normal. The selection of patients with specific disease phenotypes, the use of improved efficient drug combinations, the identification of biomarkers to specific cancer subtypes and more effective designs of investigation have improved outcomes. To move beyond the current dire condition and paucity of PC treatment options, determination of the best regimes and new treatment options is a challenge that must be met. The reasons for poor PC prognosis have remained largely unchanged for 20 years. This is arguably a consequence of significant changes in the drug discovery landscape, and the increasing pressure on academia to deliver short term 'media' friendly short-term news 'bites'. PC research sits at a pivotal point. Perhaps the greatest challenge is enacting a culture change that recognises that major breakthroughs are a result of blue sky, truly innovative and curiosity driven research.

The frequency of vitamin D-associated gene variants appear to reflect changes in long-term ultraviolet B radiation (UVB) environment, indicating interactions exist between the primary determinant of vitamin D status, UVB exposure and genetic disposition. Such interactions could have health implications, where UVB could modulate the impact of vitamin D genetic variants identified as disease risk factors. However, the current understanding of how vitamin D variants differ between populations from disparate UVB environments is limited, with previous work examining a small pool of variants and restricted populations only.Genotypic data for 46 variants within multiple vitamin D-related loci (DHCR7/NADSYN1, GC, CYP2R1, CYP11A1, CYP27A1, CYP24A1, VDR, RXRα and RXRγ) was collated from 60 sample sets (2633 subjects) with European, East Asian and Sub-Saharan African origin via the NCBI 1000 Genomes Browser and ALFRED (Allele Frequency Database), with the aim to examine for patterns in the distribution of vitamin D-associated variants across these geographic areas.The frequency of all examined genetic variants differed between populations of European, East Asian and Sub-Saharan African ancestry. Changes in the distribution of variants in CYP2R1, CYP11A1, CYP24A1, RXRα and RXRγ genes between these populations are novel findings which have not been previously reported. The distribution of several variants reflected changes in the UVB environment of the population's ancestry. However, multiple variants displayed population-specific patterns in frequency that appears not to relate to UVB changes.The reported population differences in vitamin D-related variants provides insight into the extent by which activity of the vitamin D system can differ between cohorts due to genetic variance, with potential consequences for future dietary recommendations and disease outcomes.

Mammalian sirtuins (SIRT1–7) are involved in a myriad of cellular processes, including apoptosis, proliferation, differentiation, epithelial-mesenchymal transition, aging, DNA repair, senescence, viability, survival, and stress response. In this review, we discuss the current information on the mechanistic roles of SIRT1–7 and their downstream effects (tumor promotion or suppression) in cancers of the breast and prostate. Specifically, we highlight the involvement of sirtuins in the regulation of various proteins implicated in proliferation, apoptosis, autophagy, chemoresistance, invasion, migration, and metastasis of breast and prostate cancer. Additionally, we highlight the available information regarding SIRT1–7 regulation by miRNAs, laying much emphasis on the consequences in the progression of breast and prostate cancer.

The use of edible oils and fats in dairy products is becoming increasingly important in the food industry because of their complementary functional properties. Most of these products are produced using food-grade enzymes as processing aids because processes involving enzymes are considered mild and environmentally friendly for regulatory purposes. The poor stability and recovery of enzymes in their native state limit their performance, and to enhance their activity, stability, and reusability, enzymes are often immobilised—a process that involves attaching them to a solid support. Additionally, immobilisation enables enzymes to selectively target specific substrates or products, making them highly efficient. These features have led to the increased use of immobilised enzymes in dairy and lipid processing and enzymes have been used to produce a broad range of products such as whey protein concentrates and isolates, peptide–lipid conjugates, lipid concentrates, structured lipids, and human milk fat substitutes. Therefore, this article reviews the current progress on different enzyme preparations and their use in lipid and dairy processing. It also summarises opportunities in enzyme-catalysed valorisation of dairy and lipid waste streams with the ultimate goals of sustainable food production and reductions in waste.

Proteomic techniques promise to improve the diagnosis of cholangiocarcinoma (CC) in both tissue and serum as histological diagnosis and existing serum markers exhibit poor sensitivities. We explored the use of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) to identify potential protein biomarkers of CC. Twenty-two resected CC samples were compared with adjacent noninvolved bile duct tissue. Serum from patients with CC (n=20) was compared with patients with benign disease (n=20), and healthy volunteers (n=25). Samples were analyzed on hydrophobic protein chips via SELDI-TOF MS, and classification models were developed using logistic regression and cross-validation analysis. Univariate analysis revealed 14 individual peaks differentially expressed between CC and bile duct tissue, 4 peaks between CC and benign disease, and 12 peaks between CC and sera of healthy volunteers. The 4,462 mass-to-charge serum peak had superior discriminatory ability to carbohydrate antigen 19.9 (CA19.9) and carcinoembryonic antigen (CEA) (P=.004; receiver operating characteristic [ROC] area under the curve [AUC]=0.76, 0.73, and 0.70, respectively). The training models developed panels of peaks that distinguished CC from bile duct tissue (92.5% sensitivity, 92.3% specificity; ROC AUC=0.96), CC from benign serum (65.0% sensitivity, 70.0% specificity; ROC AUC=0.83), and CC from sera of healthy volunteers (75.0% sensitivity, 100% specificity; ROC AUC=0.92). Serum results were further improved with the inclusion of CA19.9 and CEA (ROC AUC=0.86 and 0.99 for CC vs benign and healthy volunteer serum, respectively). In conclusion, biomarker panels are capable of distinguishing CC from nonmalignant tissue; serum markers have important diagnostic implications for unknown bile duct stricture.

Background & Aims: Pancreatic adenocarcinoma is a most devastating cancer that presents late and is rapidly progressive. This study aimed to identify unique, tissue-specific protein biomarkers capable of differentiating pancreatic adenocarcinoma (PC) from adjacent uninvolved pancreatic tissue (AP), benign pancreatic disease (B), and nonmalignant tumor tissue (NM). Methods: Tissue samples representing PC (n = 31), AP (n = 44), and B (n = 19) tissue were analyzed on hydrophobic protein chip arrays by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Training models were developed using logistic regression and validated using the 10-fold cross-validation approach. Results: The hydrophobic protein chip array revealed 13 protein peaks differentially expressed between PC and AP (receiver operating characteristic [ROC] area under the curve [AUC], 0.64–0.85), 8 between PC and B (ROC AUC, 0.67–0.78), and 12 between PC and NM tissue (ROC AUC, 0.63–0.81). Logistic regression and cross-validation identified overlapping panels of peaks to develop a training model that distinguished PC from AP (77.4% sensitivity, 84.1% specificity), B (83.9% sensitivity, 78.9% specificity), and NM tissue (58.1% sensitivity, 90.5% specificity). The final panels selected correctly classified 80.6% of PC and 88.6% of AP samples (ROC AUC, 0.92), 93.5% of PC and 89.5% of B samples (ROC AUC, 0.99), and 71.0% of PC and 92.1% of NM samples (ROC AUC, 0.91). Conclusions: This study used surface-enhanced laser desorption/ionization time-of-flight mass spectrometry to discover a number of protein panels that can distinguish effectively between pancreatic adenocarcinoma, benign, and adjacent pancreatic tissue. Identification of these proteins will add to our understanding of the biology of pancreatic cancer. Furthermore, these protein panels may have important diagnostic implications. Background & Aims: Pancreatic adenocarcinoma is a most devastating cancer that presents late and is rapidly progressive. This study aimed to identify unique, tissue-specific protein biomarkers capable of differentiating pancreatic adenocarcinoma (PC) from adjacent uninvolved pancreatic tissue (AP), benign pancreatic disease (B), and nonmalignant tumor tissue (NM). Methods: Tissue samples representing PC (n = 31), AP (n = 44), and B (n = 19) tissue were analyzed on hydrophobic protein chip arrays by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Training models were developed using logistic regression and validated using the 10-fold cross-validation approach. Results: The hydrophobic protein chip array revealed 13 protein peaks differentially expressed between PC and AP (receiver operating characteristic [ROC] area under the curve [AUC], 0.64–0.85), 8 between PC and B (ROC AUC, 0.67–0.78), and 12 between PC and NM tissue (ROC AUC, 0.63–0.81). Logistic regression and cross-validation identified overlapping panels of peaks to develop a training model that distinguished PC from AP (77.4% sensitivity, 84.1% specificity), B (83.9% sensitivity, 78.9% specificity), and NM tissue (58.1% sensitivity, 90.5% specificity). The final panels selected correctly classified 80.6% of PC and 88.6% of AP samples (ROC AUC, 0.92), 93.5% of PC and 89.5% of B samples (ROC AUC, 0.99), and 71.0% of PC and 92.1% of NM samples (ROC AUC, 0.91). Conclusions: This study used surface-enhanced laser desorption/ionization time-of-flight mass spectrometry to discover a number of protein panels that can distinguish effectively between pancreatic adenocarcinoma, benign, and adjacent pancreatic tissue. Identification of these proteins will add to our understanding of the biology of pancreatic cancer. Furthermore, these protein panels may have important diagnostic implications. Pancreatic cancer is one of the most devastating cancers because it presents relatively late and is rapidly progressive. In 2004, there were an estimated 32,000 new pancreatic cancer cases in the United States, with an approximately equal number of deaths.1Jemal A. Tiwari R.C. Murray T. Ghafoor A. Samuels A. Ward E. Feuer E.J. Thun M.J. Cancer statistics, 2004.CA Cancer J Clin. 2004; 54: 8-29Crossref PubMed Scopus (3932) Google Scholar Only about 20% of patients who present with clinical symptoms have a resectable tumor, leading to a 5-year survival of less than 5%.2Jemal A. Murray T. Samuels A. Ghafoor A. Ward E. Thun M.J. Cancer statistics, 2003.CA Cancer J Clin. 2003; 53: 5-26Crossref PubMed Scopus (3391) Google Scholar Consequently early detection and targeted therapy is paramount for improving a patient’s chance of survival. 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Lillemoe K.D. Cameron J.L. Yeo C.J. Hruban R.H. Goggins M. Identification of hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein I as a biomarker for pancreatic ductal adenocarcinoma by protein biochip technology.Cancer Res. 2002; 62: 1868-1875PubMed Google Scholar Recently, Chen et al42Chen R. Pan S. Brentnall T.A. Aebersold R. Proteomic profiling of pancreatic cancer for biomarker discovery.Mol Cell Proteomics. 2005; 4: 523-533Crossref PubMed Scopus (143) Google Scholar reviewed the progress and challenges for applying proteomics approaches for biomarker discovery in pancreatic cancer, and suggested that the most direct approach is to identify biomarkers in tissue. Although tissue is not as clinically accessible as blood or other body fluids, candidate biomarkers identified through proteomic profiling of tissue can provide an integral base for biomarker identification in serum. SELDI-TOF MS has the potential to provide a means of detection and identification of known proteins associated with cancer progression, and provide an opportunity for identification of novel targets associated with pancreatic cancer. This study was designed to identify unique, tissue-specific protein biomarkers capable of differentiating pancreatic adenocarcinoma from benign pancreatic and adjacent pancreatic tissue. Informed consent was obtained from a consecutive group of 50 patients, undergoing pancreatic resection for a pancreatic mass or cyst, to obtain tissue samples for a pancreatic cancer tissue bank. This protocol was approved by the Northern Sydney Health Human Research Ethics Committee (Sydney, Australia). For this study, 50 pancreatic tumor samples and 44 adjacent uninvolved pancreatic tissue (AP) samples were collected from both male (n = 22) and female (n = 28) patients. The pancreatic tumors were classified as invasive ductal pancreatic adenocarcinoma (n = 31), although none of the benign group had histologic evidence of malignant conversion (n = 19). The final histologic diagnosis of the benign group was mucinous cystadenomas (n = 7), serous cystadenomas (n = 5), solid pseudopapillary tumors (n = 2), islet cell tumors (n = 2), intraductal mucinous papillary neoplasm (n = 1), pancreatic intraepithelial neoplasia 1-A (n = 1), and chronic pancreatitis (n = 1) (Table 1).Table 1Details of Patients’ Clinical Information for Pancreatic Tumor and Adjacent Pancreatic Tissue SamplesnMean age, yRangePancreatic tumor samples5066.232–80 Male2266.347–80 Female2866.132–80Pancreatic ductal adenocarcinoma3168.439–80 Stage I1366.639–79 Stage II374.765–80 Stage III971.849–79 Stage IV661.547–79Adjacent pancreatic tissue4466.932–80Benign pancreatic tissue1962.732–80 Mucinous cystadenoma758.632–75 Serous cystadenoma568.448–80 Solid pseudopapillary tumor256.550–63 Islet cell tumor264.049–74 Intraductal mucinous papillary neoplasm170.0 Pancreatic intraepithelial neoplasia I-A165.0 Chronic pancreatitis163.0 Open table in a new tab Resected specimens were taken immediately to the pathologist where they were painted for assessment of margins, opened for selection of a representative sample of tumor and adjacent pancreatic tissue, and then snap frozen in liquid nitrogen. The time from resection to sample freezing was less than 10 minutes. Tissues were stored at −80°C until analyzed. Pancreatic adenocarcinomas were classified according to the Union Internationale Contre le Cancer TNM classification for pancreatic carcinoma.43Tsunoda T. Ura K. Eto T. Matsumoto T. Tsuchiya R. UICC and Japanese stage classifications for carcinoma of the pancreas.Int J Pancreatol. 1991; 8: 205-214PubMed Google Scholar The 31 pancreatic adenocarcinoma samples were classified into pathologic stages comprising 16 early-stage (stage I, n = 13; stage II, n = 3) and 15 late-stage (stage III, n = 9; stage IV, n = 6) pancreatic adenocarcinoma samples (Table 1). Stage IV pancreatic adenocarcinoma samples were collected as biopsy specimens from unresectable malignant tumors. Pancreatic tissue (∼50 mg) was ground to a fine powder in liquid nitrogen, solubilized by pestle homogenization in 0.6 mL of 9.5 mol/L urea/2% 3-[(3-cholamidopropyl) dimethylammonio]-1-propane-sulphate/1% dithiothreitol, then added to a QiaShredder (Qiagen, Hilden, Germany) spin column and centrifuged (12,000 rpm; 3 min) to remove insoluble material. The tissue homogenates then were reacted with a hydrophobic (H50) protein chip array surface (ProteinChip; Ciphergen Biosystems, Fremont, CA). The protein chip spots were pre-equilibrated with binding buffer (50% acetonitrile/0.5% trifluoroacetic acid) at room temperature in a humidified chamber. The samples were diluted 1:5 in binding buffer (10% acetonitrile/0.1% trifluoroacetic acid). Diluted samples (5 μL) then were applied randomly to the pre-equilibrated protein chip array spots (to assess spot-to-spot and chip-to-chip reproducibility and to reduce systematic bias) and incubated in a humidified chamber for 30 minutes at room temperature. The arrays then were washed in binding buffer (3 × 5 min), rinsed twice in Milli-Q water, and air dried. Each spot then was treated with 1 μL of a 50% saturated solution of sinapinic acid in 50% acetonitrile/0.5% trifluoroacetic acid, allowed to air-dry, then the process was repeated. The arrays then were analyzed using the Ciphergen Protein Biological System IIc ProteinChip Reader (Ciphergen Biosystems). The mass spectra of proteins were generated in the mass/charge (m/z) range of 2000–50,000 by using a laser intensity of 208–218 arbitrary units. Data were averaged from 65 spectra evenly distributed across the array spot. The laser was optimized for data collection between 5000 and 30,000 m/z, the detector sensitivity was set at 7, and peaks less than 1000 m/z were deflected away from the detector. The m/z value for each of the peaks was determined using external calibration with known standards (Sigma, St Louis, MO): bovine insulin (5734.51 +1H), equine cytochrome c (12,361.96 +1H), equine apomyoglobin (16,952.27 +1H), and rabbit muscle aldolase (39,212.28 +1H). All data were analyzed using the Ciphergen ProteinChip Software version 3.1 (Ciphergen Biosystems). The raw peak intensity data for comparison of each spectrum was normalized using the total ion current between 2500 and 20,000 m/z. The detection of peaks differentially expressed between each group was performed using the Biomarker Wizard utility (version 3.1; Ciphergen Biosystems) and sample group statistics were performed for profiles of adjacent pancreatic tissue vs adenocarcinoma tissue, adjacent pancreatic tissue vs benign tissue, and adenocarcinoma vs benign tissue. Univariate analysis was performed between groups using the Mann–Whitney U test and results were considered significantly different when the P value was less than .05. For each putative marker, receiver operating characteristic (ROC) curves were generated to evaluate their discriminatory power (SPSS Software version 13.0; SPSS, Chicago, IL). Training models were developed using multivariate binary logistic regression to determine which peaks were able to best predict pancreatic adenocarcinoma from adjacent pancreatic tissue, benign pancreatic (B) tissue, and nonmalignant (NM) tissue (combination of B and AP tissue). For each comparison analyzed (ie, pancreatic adenocarcinoma [PC] vs AP, PC vs B, and PC vs NM), the models were validated using the 10-fold cross-validation approach as described by Ambroise and McLachlan.44Ambroise C. McLachlan G.J. Selection bias in gene extraction on the basis of microarray gene-expression data.Proc Natl Acad Sci U S A. 2002; 99: 6562-6566Crossref PubMed Scopus (1109) Google Scholar This repeated random sampling procedure allows for the use of all samples within the dataset to be tested and for the correction of any selection bias. Briefly, the 10-fold cross-validation approach divides the entire randomized dataset into 10 nonoverlapping datasets of roughly equal size. The model is trained on 9 of these subsets and then tested on the remaining subset to obtain prediction values. This process is repeated in turn for each of the remaining subsets, each of which leads to a different model. So, for each subset, the different model is tested on an independent sample that was not included in the development of the training model. This enabled the calculation of unbiased estimates of sensitivity and specificity, overall accuracy, and ROC area under the curve (AUC) values of the candidate tumor biomarker panels. As Ambroise and McLachlan44Ambroise C. McLachlan G.J. Selection bias in gene extraction on the basis of microarray gene-expression data.Proc Natl Acad Sci U S A. 2002; 99: 6562-6566Crossref PubMed Scopus (1109) Google Scholar stressed, in obtaining unbiased estimates it is important to avoid selection bias, namely to allow the cross-validation of the prediction rule to be external to the selection process. The effect of accommodating the selection bias is that different peaks may be selected in each training model. This is a direct consequence of the test set playing no part in the selection of the peaks. To assess whether the data within the logistic model were a good fit, the Hosmer and Lemeshow Goodness of Fit test was applied (SPSS software version 13.0). This external cross-validation approach ensures that for each subset the test samples were not included in the development of the training models. Logistic regression also was used to assess whether a panel of proteins could subclassify pancreatic adenocarcinoma successfully into early (stage I/II) vs late (stage III/IV) stage disease. Samples were analyzed in duplicate to allow for calculation of the coefficient of variation for peak intensity and mass accuracy. Replicate peak intensity values generated by SELDI-TOF MS then were averaged. Twelve peaks across the spectrum were used for the estimation of coefficient of variations. These peaks were selected randomly from all common peaks and included individual peaks that were expressed differentially by univariate analysis and peaks not significantly different between groups. The peaks included in the estimation of coefficient of variations possessed a wide distribution of peak intensities. This was performed across the repeated spectra and displayed as an overall mean. The coefficient of variation values in our study (peak intensity, 12%–20%; mass accuracy, 0.01%–0.03%) compare favorably with coefficient of variations reported in previous SELDI studies,16Kozak K.R. Amneus M.W. Pusey S.M. Su F. Luong M.N. Luong S.A. Reddy S.T. Farias-Eisner R. Identification of biomarkers for ovarian cancer using strong anion-exchange ProteinChips potential use in diagnosis and prognosis.Proc Natl Acad Sci U S A. 2003; 100: 12343-12348Crossref PubMed Scopus (240) Google Scholar, 18Petricoin E.F. Ardekani A.M. Hitt B.A. Levine P.J. Fusaro V.A. Steinberg S.M. Mills G.B. Simone C. Fishman D.A. Kohn E.C. Liotta L.A. Use of proteomic patterns in serum to identify ovarian cancer.Lancet. 2002; 359: 572-577Abstract Full Text Full Text PDF PubMed Scopus (2889) Google Scholar, 39Koopmann J. Zhang Z. White N. Rosenzweig J. Fedarko N. Jagannath S. Canto M.I. Yeo C.J. Chan D.W. Goggins M. Serum diagnosis of pancreatic adenocarcinoma using surface-enhanced laser desorption and ionization mass spectrometry.Clin Cancer Res. 2004; 10: 860-868Crossref PubMed Scopus (266) Google Scholar, 45Wadsworth J.T. Somers K.D. Cazares L.H. Malik G. Adam B.L. Stack Jr, B.C. Wright Jr, G.L. Semmes O.J. Serum protein profiles to identify head and neck cancer.Clin Cancer Res. 2004; 10: 1625-1632Crossref PubMed Scopus (106) Google Scholar, 46Li J. Zhang Z. Rosenzweig J. Wang Y.Y. Chan D.W. Proteomics and bioinformatics approaches for identification of serum biomarkers to detect breast cancer.Clin Chem. 2002; 48: 1296-1304PubMed Google Scholar and accuracy might be improved with the use of automation. Protein chip technology coupled with SELDI-TOF MS showed clear differences in protein profile expressions between invasive pancreatic adenocarcinoma, benign, and adjacent pancreatic tissue using the H50 array. An example from a segment of the protein mass profile between 9700 and 12,400 m/z is shown in Figure 1, accompanied by a spectral overlay of these markers. The H50 array revealed 13 protein peaks that were expressed differentially between invasive pancreatic adenocarcinoma vs adjacent pancreatic tissue. These individual putative tumor markers had ROC AUC values ranging from 0.64 to 0.85 (Table 2). Of these 13 protein peaks, 8 were up-regulated in the pancreatic adenocarcinoma group.Table 2Protein Peaks Expressed Differentially Between Pancreatic Adenocarcinoma Tissue Vs Adjacent Pancreatic Tissue, Benign Pancreatic Tissue, and Nonmalignant TissuePancreatic adenocarcinoma

Myc oncoproteins and histone deacetylases (HDACs) exert oncogenic effects by modulating gene transcription.Paradoxically, N-Myc induces p53 gene expression.Tumor protein 53-induced nuclear protein 1 (TP53INP1) phosphorylates p53 protein at serine 46, leading to enhanced p53 activity, transcriptional activation of p53 target genes and programmed cell death.Here we aimed to identify the mechanism through which N-Myc overexpressing p53 wild-type neuroblastoma cells acquired resistance to apoptosis.TP53INP1 was found to be one of the genes most significantly repressed by HDAC2 and N-Myc according to Affymetrix microarray gene expression datasets.HDAC2 and N-Myc reduced TP53INP1 gene expression by direct binding to the TP53INP1 gene promoter, leading to transcriptional repression of TP53INP1, p53 protein de-phosphorylation at serine 46, neuroblastoma cell proliferation and survival.Moreover, low levels of TP53INP1 expression in human neuroblastoma tissues correlated with high levels of N-Myc expression and poor patient outcome, and the BET bromodomain inhibitors JQ1 and I-BET151 reduced N-Myc expression and reactivated TP53INP1 expression in neuroblastoma cells.These findings identify TP53INP1 repression as an important co-factor for N-Myc oncogenesis, and provide further evidence for the potential application of BET bromodomain inhibitors in the therapy of N-Myc-induced neuroblastoma.

This study examined the effects of different ratios of shellac (20–60%), stearic acid (SA) (0–2%), and Tween-20 (0.1–0.5 ml) on the water vapor permeability (WVP) and mechanical properties of the pea starch-guar gum (PSGG) films which were evaluated by using response surface methodology (RSM). The incorporation of shellac into the PSGG film structure led to a slightly increased of film thickness. However the addition of higher concentrations of shellac did not improve the moisture barrier of PSGG film owing to the poor distribution of shellac in the film structure. Film formulated with 40% shellac, 1% SA, and 0.3% Tween-20 exhibited optimal functional properties. Moreover, the influence of the incorporation of different emulsifiers into the optimized film matrix was investigated by studying the physical, mechanical, and optical properties of the films. Films containing oleic acid (OA) showed not only lower thickness, WVP, moisture content, and water solubility, but also higher percentage of elongation (E), tensile strength (TS), and transparency compared with other fatty acids tested. Biocomposite pea starch-guar gum-shellac (PSGG-Sh) films containing OA can be considered to be sufficient for most of food packaging applications.

Pancreatic cancer is an inherently aggressive disease with an extremely poor prognosis and lack of effective treatments. Over the past few decades, much has been uncovered regarding the pathogenesis of pancreatic cancer and the underlying genetic alterations necessary for tumour initiation and progression. Much of what we know about pancreatic cancer has come from mouse models of this disease. This review focusses on the development of genetically engineered mouse models that phenotypically and genetically recapitulate human pancreatic cancer, as well as the increasing use of patient-derived xenografts for preclinical studies and the development of personalised medicine strategies.

Catharanthus roseus (L.) G. Don (C. roseus) is well known as an important medicinal plant, with compounds such as the vinca alkaloids isolated for their anticancer activity. As such, it is important to determine the effective solvent for bioactive compound extraction from this plant and the suitable drying methods for preparation of starting material. The aims of this study were to determine the effect of extraction solvents and drying methods on bioactive compounds and antioxidant properties of C. roseus. Water was found to be the optimal solvent for phenolic and flavonoid extraction; whereas, methanol was the best solvent for saponin and proanthocyanidin extraction. The data also revealed that vacuum drying at 50°C was suitable for drying the leaf and the flower which contained high levels of phenolics and flavonoids, while infrared drying at 35°C was recommended for drying the stem and the root which had high saponin content. Practical applications This study suggested that different parts of C. roseus had different suitable thermal drying methods. For the leaf and the flower, vacuum drying at 50°C was the optimal drying method, whereas infrared drying at 35°C was suitable for drying the stem and the root. These drying conditions can be easily applied for preparation of dried plant parts with high levels of bioactive compounds in the large scale. Importantly, the data indicated that the stem and the root of C. roseus which were considered as waste when the leaf was used for exploiting alkaloids, possessed great content of saponins. Therefore, these parts can be further used for isolation and purification of saponins.

This data article indicates the in vitro cytotoxicity of kaempferol and gallic acid across different cancer cell lines including A2780 (ovarian), H460 (lung), A431 (skin), MIA PaCa-2 (pancreas), Du145 (prostate), HT29 (colon), MCF-7 (breast), BE2-C (neuroblastoma), SJ-G2, U87 and SMA (glioblastoma). The dataset showed that the inhibitory activity of kaempferol was comparatively stronger than gallic acid. Thereby, kaempferol is offered as a potent anticancer agent for further investigation and beneficial as a dietary supplement. The data within this article relates to the research article entitled “Screening phytochemical content, antioxidant, antimicrobial and cytotoxic activities of Catharanthus roseus (L.) G. Don stem extract and its fractions” (Pham et al., 2018).