Sample records for factor ripening inhibitor

  1. The regulatory mechanism of fruit ripening revealed by analyses of direct targets of the tomato MADS-box transcription factor RIPENING INHIBITOR

    PubMed Central

    Fujisawa, Masaki; Ito, Yasuhiro

    2013-01-01

    The developmental process of ripening is unique to fleshy fruits and a key factor in fruit quality. The tomato (Solanum lycopersicum) MADS-box transcription factor RIPENING INHIBITOR (RIN), one of the earliest-acting ripening regulators, is required for broad aspects of ripening, including ethylene-dependent and -independent pathways. However, our knowledge of direct RIN target genes has been limited, considering the broad effects of RIN on ripening. In a recent work published in The Plant Cell, we identified 241 direct RIN target genes by chromatin immunoprecipitation coupled with DNA microarray (ChIP-chip) and transcriptome analysis. Functional classification of the targets revealed that RIN participates in the regulation of many biological processes including well-known ripening processes such as climacteric ethylene production and lycopene accumulation. In addition, we found that ethylene is required for the full expression of RIN and several RIN-targeting transcription factor genes at the ripening stage. Here, based on our recently published findings and additional data, we discuss the ripening processes regulated by RIN and the interplay between RIN and ethylene. PMID:23518588

  2. Comprehensive Profiling of Ethylene Response Factor Expression Identifies Ripening-Associated ERF Genes and Their Link to Key Regulators of Fruit Ripening in Tomato1[OPEN

    PubMed Central

    Gomes, Bruna Lima; Mila, Isabelle; Frasse, Pierre; Zouine, Mohamed; Bouzayen, Mondher

    2016-01-01

    Our knowledge of the factors mediating ethylene-dependent ripening of climacteric fruit remains limited. The transcription of ethylene-regulated genes is mediated by ethylene response factors (ERFs), but mutants providing information on the specific role of the ERFs in fruit ripening are still lacking, likely due to functional redundancy among this large multigene family of transcription factors. We present here a comprehensive expression profiling of tomato (Solanum lycopersicum) ERFs in wild-type and tomato ripening-impaired tomato mutants (Never-ripe [Nr], ripening-inhibitor [rin], and non-ripening [nor]), indicating that out of the 77 ERFs present in the tomato genome, 27 show enhanced expression at the onset of ripening while 28 display a ripening-associated decrease in expression, suggesting that different ERFs may have contrasting roles in fruit ripening. Among the 19 ERFs exhibiting the most consistent up-regulation during ripening, the expression of 11 ERFs is strongly down-regulated in rin, nor, and Nr tomato ripening mutants, while only three are consistently up-regulated. Members of subclass E, SlERF.E1, SlERF.E2, and SlERF.E4, show dramatic down-regulation in the ripening mutants, suggesting that their expression might be instrumental in fruit ripening. This study illustrates the high complexity of the regulatory network connecting RIN and ERFs and identifies subclass E members as the most active ERFs in ethylene- and RIN/NOR-dependent ripening. PMID:26739234

  3. Effect of ripening inhibitor type on formation, stability, and antimicrobial activity of thyme oil nanoemulsion.

    PubMed

    Ryu, Victor; McClements, David J; Corradini, Maria G; McLandsborough, Lynne

    2018-04-15

    The objective of this research was to study the impact of ripening inhibitor level and type on the formation, stability, and activity of antimicrobial thyme oil nanoemulsions formed by spontaneous emulsification. Oil-in-water antimicrobial nanoemulsions (10 wt%) were formed by titrating a mixture of essential oil, ripening inhibitor, and surfactant (Tween 80) into 5 mM sodium citrate buffer (pH 3.5). Stable nanoemulsions containing small droplets (d < 70 nm) were formed. The antimicrobial activity of the nanoemulsions decreased with increasing ripening inhibitor concentration which was attributed to a reduction in the amount of hydrophobic antimicrobial constituents transferred to the separated hydrophobic domain, mimicking bacterial cell membranes, by using dialysis and chromatography. The antimicrobial activity of the nanoemulsions also depended on the nature of the ripening inhibitor used: palm ≈ corn > canola > coconut which also depended on their ability to transfer hydrophobic antimicrobial constituents to the separated hydrophobic domain. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Fruit ripening mutants reveal cell metabolism and redox state during ripening.

    PubMed

    Kumar, Vinay; Irfan, Mohammad; Ghosh, Sumit; Chakraborty, Niranjan; Chakraborty, Subhra; Datta, Asis

    2016-03-01

    Ripening which leads to fruit senescence is an inimitable process characterized by vivid changes in color, texture, flavor, and aroma of the fleshy fruits. Our understanding of the mechanisms underlying the regulation of fruit ripening and senescence is far from complete. Molecular and biochemical studies on tomato (Solanum lycopersicum) ripening mutants such as ripening inhibitor (rin), nonripening (nor), and never ripe (Nr) have been useful in our understanding of fruit development and ripening. The MADS-box transcription factor RIN, a global regulator of fruit ripening, is vital for the broad aspects of ripening, in both ethylene-dependent and independent manners. Here, we have carried out microarray analysis to study the expression profiles of tomato genes during ripening of wild type and rin mutant fruits. Analysis of the differentially expressed genes revealed the role of RIN in regulation of several molecular and biochemical events during fruit ripening including fruit specialized metabolism and cellular redox state. The role of reactive oxygen species (ROS) during fruit ripening and senescence was further examined by determining the changes in ROS level during ripening of wild type and mutant fruits and by analyzing expression profiles of the genes involved in maintaining cellular redox state. Taken together, our findings suggest an important role of ROS during fruit ripening and senescence, and therefore, modulation of ROS level during ripening could be useful in achieving desired fruit quality.

  5. Abscisic acid and sucrose regulate tomato and strawberry fruit ripening through the abscisic acid-stress-ripening transcription factor.

    PubMed

    Jia, Haifeng; Jiu, Songtao; Zhang, Cheng; Wang, Chen; Tariq, Pervaiz; Liu, Zhongjie; Wang, Baoju; Cui, Liwen; Fang, Jinggui

    2016-10-01

    Although great progress has been made towards understanding the role of abscisic acid (ABA) and sucrose in fruit ripening, the mechanisms underlying the ABA and sucrose signalling pathways remain elusive. In this study, transcription factor ABA-stress-ripening (ASR), which is involved in the transduction of ABA and sucrose signalling pathways, was isolated and analysed in the nonclimacteric fruit, strawberry and the climacteric fruit, tomato. We have identified four ASR isoforms in tomato and one in strawberry. All ASR sequences contained the ABA stress- and ripening-induced proteins and water-deficit stress-induced proteins (ABA/WDS) domain and all ASR transcripts showed increased expression during fruit development. The expression of the ASR gene was influenced not only by sucrose and ABA, but also by jasmonic acid (JA) and indole-3-acetic acid (IAA), and these four factors were correlated with each other during fruit development. ASR bound the hexose transporter (HT) promoter, which contained a sugar box that activated downstream gene expression. Overexpression of the ASR gene promoted fruit softening and ripening, whereas RNA interference delayed fruit ripening, as well as affected fruit physiological changes. Change in ASR gene expression influenced the expression of several ripening-related genes such as CHS, CHI, F3H, DFR, ANS, UFGT, PG, PL, EXP1/2, XET16, Cel1/2 and PME. Taken together, this study may provide new evidence on the important role of ASR in cross-signalling between ABA and sucrose to regulate tomato and strawberry fruit ripening. The findings of this study also provide new insights into the regulatory mechanism underlying fruit development. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  6. Factors affecting skin tannin extractability in ripening grapes.

    PubMed

    Bindon, Keren A; Madani, S Hadi; Pendleton, Phillip; Smith, Paul A; Kennedy, James A

    2014-02-05

    The acetone-extractable (70% v/v) skin tannin content of Vitis vinifera L. cv. Cabernet Sauvignon grapes was found to increase during late-stage ripening. Conversely, skin tannin content determined following ethanol extraction (10, 20, and 50% v/v) did not consistently reflect this trend. The results indicated that a fraction of tannin became less extractable in aqueous ethanol during ripening. Skin cell walls were observed to become more porous during ripening, which may facilitate the sequestering of tannin as an adsorbed fraction within cell walls. For ethanol extracts, tannin molecular mass increased with advancing ripeness, even when extractable tannin content was constant, but this effect was negligible in acetone extracts. Reconstitution experiments with isolated skin tannin and cell wall material indicated that the selectivity of tannin adsorption by cell walls changed as tannin concentration increased. Tannin concentration, tannin molecular mass, and cell wall porosity are discussed as factors that may influence skin tannin extractability.

  7. Prostaglandin dehydrogenase is a target for successful induction of cervical ripening

    PubMed Central

    Kishore, Annavarapu Hari; Liang, Hanquan; Xing, Chao; Ganesh, Thota; Akgul, Yucel; Posner, Bruce; Ready, Joseph M.; Markowitz, Sanford D.; Word, Ruth Ann

    2017-01-01

    The cervix represents a formidable structural barrier for successful induction of labor. Approximately 10% of pregnancies undergo induction of cervical ripening and labor with prostaglandin (PG) E2 or PGE analogs, often requiring many hours of hospitalization and monitoring. On the other hand, preterm cervical ripening in the second trimester predicts preterm birth. The regulatory mechanisms of this paradoxical function of the cervix are unknown. Here, we show that PGE2 uses cell-specific EP2 receptor-mediated increases in Ca2+ to dephosphorylate and translocate histone deacetylase 4 (HDAC4) to the nucleus for repression of 15-hydroxy prostaglandin dehydrogenase (15-PGDH). The crucial role of 15-PGDH in cervical ripening was confirmed in vivo. Although PGE2 or 15-PGDH inhibitor alone did not alter gestational length, treatment with 15-PGDH inhibitor + PGE2 or metabolism-resistant dimethyl-PGE2 resulted in preterm cervical ripening and delivery in mice. The ability of PGE2 to selectively autoamplify its own synthesis in stromal cells by signaling transcriptional repression of 15-PGDH elucidates long sought-after molecular mechanisms that govern PG action in the cervix. This report details unique mechanisms of action in the cervix and serves as a catalyst for (i) the use of 15-PGDH inhibitors to initiate or amplify low-dose PGE2-mediated cervical ripening or (ii) EP2 receptor antagonists, HDAC4 inhibitors, and 15-PGDH activators to prevent preterm cervical ripening and preterm birth. PMID:28716915

  8. Ethylene-induced transcriptional and hormonal responses at the onset of sugarcane ripening

    PubMed Central

    Cunha, Camila P.; Roberto, Guilherme G.; Vicentini, Renato; Lembke, Carolina G.; Souza, Glaucia M.; Ribeiro, Rafael V.; Machado, Eduardo C.; Lagôa, Ana M. M. A.; Menossi, Marcelo

    2017-01-01

    The effects of ethephon as a sugarcane ripener are attributed to ethylene. However, the role of this phytohormone at the molecular level is unknown. We performed a transcriptome analysis combined with the evaluation of sucrose metabolism and hormone profiling of sugarcane plants sprayed with ethephon or aminoethoxyvinylglycine (AVG), an ethylene inhibitor, at the onset of ripening. The differential response between ethephon and AVG on sucrose level and sucrose synthase activity in internodes indicates ethylene as a potential regulator of sink strength. The correlation between hormone levels and transcriptional changes suggests ethylene as a trigger of multiple hormone signal cascades, with approximately 18% of differentially expressed genes involved in hormone biosynthesis, metabolism, signalling, and response. A defence response elicited in leaves favoured salicylic acid over the ethylene/jasmonic acid pathway, while the upper internode was prone to respond to ethylene with strong stimuli on ethylene biosynthesis and signalling genes. Besides, ethylene acted synergistically with abscisic acid, another ripening factor, and antagonistically with gibberellin and auxin. We identified potential ethylene target genes and characterized the hormonal status during ripening, providing insights into the action of ethylene at the site of sucrose accumulation. A molecular model of ethylene interplay with other hormones is proposed. PMID:28266527

  9. Tomato fruit chromoplasts behave as respiratory bioenergetic organelles during ripening.

    PubMed

    Renato, Marta; Pateraki, Irini; Boronat, Albert; Azcón-Bieto, Joaquín

    2014-10-01

    During tomato (Solanum lycopersicum) fruit ripening, chloroplasts differentiate into photosynthetically inactive chromoplasts. It was recently reported that tomato chromoplasts can synthesize ATP through a respiratory process called chromorespiration. Here we show that chromoplast oxygen consumption is stimulated by the electron donors NADH and NADPH and is sensitive to octyl gallate (Ogal), a plastidial terminal oxidase inhibitor. The ATP synthesis rate of isolated chromoplasts was dependent on the supply of NAD(P)H and was fully inhibited by Ogal. It was also inhibited by the proton uncoupler carbonylcyanide m-chlorophenylhydrazone, suggesting the involvement of a chemiosmotic gradient. In addition, ATP synthesis was sensitive to 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, a cytochrome b6f complex inhibitor. The possible participation of this complex in chromorespiration was supported by the detection of one of its components (cytochrome f) in chromoplasts using immunoblot and immunocytochemical techniques. The observed increased expression of cytochrome c6 during ripening suggests that it could act as electron acceptor of the cytochrome b6f complex in chromorespiration. The effects of Ogal on respiration and ATP levels were also studied in tissue samples. Oxygen uptake of mature green fruit and leaf tissues was not affected by Ogal, but was inhibited increasingly in fruit pericarp throughout ripening (up to 26% in red fruit). Similarly, Ogal caused a significant decrease in ATP content of red fruit pericarp. The number of energized mitochondria, as determined by confocal microscopy, strongly decreased in fruit tissue during ripening. Therefore, the contribution of chromoplasts to total fruit respiration appears to increase in late ripening stages. © 2014 American Society of Plant Biologists. All Rights Reserved.

  10. Transcriptome Analysis Provides a Preliminary Regulation Route of the Ethylene Signal Transduction Component, SlEIN2, during Tomato Ripening.

    PubMed

    Wang, Rui-Heng; Yuan, Xin-Yu; Meng, Lan-Huan; Zhu, Ben-Zhong; Zhu, Hong-Liang; Luo, Yun-Bo; Fu, Da-Qi

    2016-01-01

    Ethylene is crucial in climacteric fruit ripening. The ethylene signal pathway regulates several physiological alterations such as softening, carotenoid accumulation and sugar level reduction, and production of volatile compounds. All these physiological processes are controlled by numerous genes and their expression simultaneously changes at the onset of ripening. Ethylene insensitive 2 (EIN2) is a key component for ethylene signal transduction, and its mutation causes ethylene insensitivity. In tomato, silencing SlEIN2 resulted in a non-ripening phenotype and low ethylene production. RNA sequencing of SlEIN2-silenced and wild type tomato, and differential gene expression analyses, indicated that silencing SlEIN2 caused changes in more than 4,000 genes, including those related to photosynthesis, defense, and secondary metabolism. The relative expression level of 28 genes covering ripening-associated transcription factors, ethylene biosynthesis, ethylene signal pathway, chlorophyll binding proteins, lycopene and aroma biosynthesis, and defense pathway, showed that SlEIN2 influences ripening inhibitor (RIN) in a feedback loop, thus controlling the expression of several other genes. SlEIN2 regulates many aspects of fruit ripening, and is a key factor in the ethylene signal transduction pathway. Silencing SlEIN2 ultimately results in lycopene biosynthesis inhibition, which is the reason why tomato does not turn red, and this gene also affects the expression of several defense-associated genes. Although SlEIN2-silenced and green wild type fruits are similar in appearance, their metabolism is significantly different at the molecular level.

  11. Banana ethylene response factors are involved in fruit ripening through their interactions with ethylene biosynthesis genes.

    PubMed

    Xiao, Yun-yi; Chen, Jian-ye; Kuang, Jiang-fei; Shan, Wei; Xie, Hui; Jiang, Yue-ming; Lu, Wang-jin

    2013-05-01

    The involvement of ethylene response factor (ERF) transcription factor (TF) in the transcriptional regulation of ethylene biosynthesis genes during fruit ripening remains largely unclear. In this study, 15 ERF genes, designated as MaERF1-MaERF15, were isolated and characterized from banana fruit. These MaERFs were classified into seven of the 12 known ERF families. Subcellular localization showed that MaERF proteins of five different subfamilies preferentially localized to the nucleus. The 15 MaERF genes displayed differential expression patterns and levels in peel and pulp of banana fruit, in association with four different ripening treatments caused by natural, ethylene-induced, 1-methylcyclopropene (1-MCP)-delayed, and combined 1-MCP and ethylene treatments. MaERF9 was upregulated while MaERF11 was downregulated in peel and pulp of banana fruit during ripening or after treatment with ethylene. Furthermore, yeast-one hybrid (Y1H) and transient expression assays showed that the potential repressor MaERF11 bound to MaACS1 and MaACO1 promoters to suppress their activities and that MaERF9 activated MaACO1 promoter activity. Interestingly, protein-protein interaction analysis revealed that MaERF9 and -11 physically interacted with MaACO1. Taken together, these results suggest that MaERFs are involved in banana fruit ripening via transcriptional regulation of or interaction with ethylene biosynthesis genes.

  12. Banana ethylene response factors are involved in fruit ripening through their interactions with ethylene biosynthesis genes

    PubMed Central

    Xiao, Yun-yi; Chen, Jian-ye; Kuang, Jiang-fei; Shan, Wei; Xie, Hui; Jiang, Yue-ming; Lu, Wang-jin

    2013-01-01

    The involvement of ethylene response factor (ERF) transcription factor (TF) in the transcriptional regulation of ethylene biosynthesis genes during fruit ripening remains largely unclear. In this study, 15 ERF genes, designated as MaERF1–MaERF15, were isolated and characterized from banana fruit. These MaERFs were classified into seven of the 12 known ERF families. Subcellular localization showed that MaERF proteins of five different subfamilies preferentially localized to the nucleus. The 15 MaERF genes displayed differential expression patterns and levels in peel and pulp of banana fruit, in association with four different ripening treatments caused by natural, ethylene-induced, 1-methylcyclopropene (1-MCP)-delayed, and combined 1-MCP and ethylene treatments. MaERF9 was upregulated while MaERF11 was downregulated in peel and pulp of banana fruit during ripening or after treatment with ethylene. Furthermore, yeast-one hybrid (Y1H) and transient expression assays showed that the potential repressor MaERF11 bound to MaACS1 and MaACO1 promoters to suppress their activities and that MaERF9 activated MaACO1 promoter activity. Interestingly, protein–protein interaction analysis revealed that MaERF9 and -11 physically interacted with MaACO1. Taken together, these results suggest that MaERFs are involved in banana fruit ripening via transcriptional regulation of or interaction with ethylene biosynthesis genes. PMID:23599278

  13. Temperature and relative humidity influence the ripening descriptors of Camembert-type cheeses throughout ripening.

    PubMed

    Leclercq-Perlat, M-N; Sicard, M; Perrot, N; Trelea, I C; Picque, D; Corrieu, G

    2015-02-01

    Ripening descriptors are the main factors that determine consumers' preferences of soft cheeses. Six descriptors were defined to represent the sensory changes in Camembert cheeses: Penicillium camemberti appearance, cheese odor and rind color, creamy underrind thickness and consistency, and core hardness. To evaluate the effects of the main process parameters on these descriptors, Camembert cheeses were ripened under different temperatures (8, 12, and 16°C) and relative humidity (RH; 88, 92, and 98%). The sensory descriptors were highly dependent on the temperature and RH used throughout ripening in a ripening chamber. All sensory descriptor changes could be explained by microorganism growth, pH, carbon substrate metabolism, and cheese moisture, as well as by microbial enzymatic activities. On d 40, at 8°C and 88% RH, all sensory descriptors scored the worst: the cheese was too dry, its odor and its color were similar to those of the unripe cheese, the underrind was driest, and the core was hardest. At 16°C and 98% RH, the odor was strongly ammonia and the color was dark brown, and the creamy underrind represented the entire thickness of the cheese but was completely runny, descriptors indicative of an over ripened cheese. Statistical analysis showed that the best ripening conditions to achieve an optimum balance between cheese sensory qualities and marketability were 13±1°C and 94±1% RH. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. Comprehensive Analysis of ABA Effects on Ethylene Biosynthesis and Signaling during Tomato Fruit Ripening.

    PubMed

    Mou, Wangshu; Li, Dongdong; Bu, Jianwen; Jiang, Yuanyuan; Khan, Zia Ullah; Luo, Zisheng; Mao, Linchun; Ying, Tiejin

    2016-01-01

    ABA has been widely acknowledged to regulate ethylene biosynthesis and signaling during fruit ripening, but the molecular mechanism underlying the interaction between these two hormones are largely unexplored. In the present study, exogenous ABA treatment obviously promoted fruit ripening as well as ethylene emission, whereas NDGA (Nordihydroguaiaretic acid, an inhibitor of ABA biosynthesis) application showed the opposite biological effects. Combined RNA-seq with time-course RT-PCR analysis, our study not only helped to illustrate how ABA regulated itself at the transcription level, but also revealed that ABA can facilitate ethylene production and response probably by regulating some crucial genes such as LeACS4, LeACO1, GR and LeETR6. In addition, investigation on the fruits treated with 1-MCP immediately after ABA exposure revealed that ethylene might be essential for the induction of ABA biosynthesis and signaling at the onset of fruit ripening. Furthermore, some specific transcription factors (TFs) known as regulators of ethylene synthesis and sensibility (e.g. MADS-RIN, TAGL1, CNR and NOR) were also observed to be ABA responsive, which implied that ABA influenced ethylene action possibly through the regulation of these TFs expression. Our comprehensive physiological and molecular-level analysis shed light on the mechanism of cross-talk between ABA and ethylene during the process of tomato fruit ripening.

  15. Comprehensive Analysis of ABA Effects on Ethylene Biosynthesis and Signaling during Tomato Fruit Ripening

    PubMed Central

    Bu, Jianwen; Jiang, Yuanyuan; Khan, Zia Ullah; Luo, Zisheng; Mao, Linchun; Ying, Tiejin

    2016-01-01

    ABA has been widely acknowledged to regulate ethylene biosynthesis and signaling during fruit ripening, but the molecular mechanism underlying the interaction between these two hormones are largely unexplored. In the present study, exogenous ABA treatment obviously promoted fruit ripening as well as ethylene emission, whereas NDGA (Nordihydroguaiaretic acid, an inhibitor of ABA biosynthesis) application showed the opposite biological effects. Combined RNA-seq with time-course RT-PCR analysis, our study not only helped to illustrate how ABA regulated itself at the transcription level, but also revealed that ABA can facilitate ethylene production and response probably by regulating some crucial genes such as LeACS4, LeACO1, GR and LeETR6. In addition, investigation on the fruits treated with 1-MCP immediately after ABA exposure revealed that ethylene might be essential for the induction of ABA biosynthesis and signaling at the onset of fruit ripening. Furthermore, some specific transcription factors (TFs) known as regulators of ethylene synthesis and sensibility (e.g. MADS-RIN, TAGL1, CNR and NOR) were also observed to be ABA responsive, which implied that ABA influenced ethylene action possibly through the regulation of these TFs expression. Our comprehensive physiological and molecular-level analysis shed light on the mechanism of cross-talk between ABA and ethylene during the process of tomato fruit ripening. PMID:27100326

  16. Tomato GOLDEN2-LIKE Transcription Factors Reveal Molecular Gradients That Function during Fruit Development and Ripening[W][OPEN

    PubMed Central

    Nguyen, Cuong V.; Vrebalov, Julia T.; Gapper, Nigel E.; Zheng, Yi; Zhong, Silin; Fei, Zhangjun; Giovannoni, James J.

    2014-01-01

    Fruit ripening is the summation of changes rendering fleshy fruit tissues attractive and palatable to seed dispersing organisms. For example, sugar content is influenced by plastid numbers and photosynthetic activity in unripe fruit and later by starch and sugar catabolism during ripening. Tomato fruit are sinks of photosynthate, yet unripe green fruit contribute significantly to the sugars that ultimately accumulate in the ripe fruit. Plastid numbers and chlorophyll content are influenced by numerous environmental and genetic factors and are positively correlated with photosynthesis and photosynthate accumulation. GOLDEN2-LIKE (GLK) transcription factors regulate plastid and chlorophyll levels. Tomato (Solanum lycopersicum), like most plants, contains two GLKs (i.e., GLK1 and GLK2/UNIFORM). Mutant and transgene analysis demonstrated that these genes encode functionally similar peptides, though differential expression renders GLK1 more important in leaves, while GLK2 is predominant in fruit. A latitudinal gradient of GLK2 expression influences the typical uneven coloration of green and ripe wild-type fruit. Transcriptome profiling revealed a broader fruit gene expression gradient throughout development. The gradient influenced general ripening activities beyond plastid development and was consistent with the easily observed yet poorly studied ripening gradient present in tomato and many fleshy fruits. PMID:24510723

  17. Isolation of ripening-related genes from ethylene/1-MCP treated papaya through RNA-seq.

    PubMed

    Shen, Yan Hong; Lu, Bing Guo; Feng, Li; Yang, Fei Ying; Geng, Jiao Jiao; Ming, Ray; Chen, Xiao Jing

    2017-08-31

    Since papaya is a typical climacteric fruit, exogenous ethylene (ETH) applications can induce premature and quicker ripening, while 1-methylcyclopropene (1-MCP) slows down the ripening processes. Differential gene expression in ETH or 1-MCP-treated papaya fruits accounts for the ripening processes. To isolate the key ripening-related genes and better understand fruit ripening mechanisms, transcriptomes of ETH or 1-MCP-treated, and non-treated (Control Group, CG) papaya fruits were sequenced using Illumina Hiseq2500. A total of 18,648 (1-MCP), 19,093 (CG), and 15,321 (ETH) genes were detected, with the genes detected in the ETH-treatment being the least. This suggests that ETH may inhibit the expression of some genes. Based on the differential gene expression (DGE) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, 53 fruit ripening-related genes were selected: 20 cell wall-related genes, 18 chlorophyll and carotenoid metabolism-related genes, four proteinases and their inhibitors, six plant hormone signal transduction pathway genes, four transcription factors, and one senescence-associated gene. Reverse transcription quantitative PCR (RT-qPCR) analyses confirmed the results of RNA-seq and verified that the expression pattern of six genes is consistent with the fruit senescence process. Based on the expression profiling of genes in carbohydrate metabolic process, chlorophyll metabolism pathway, and carotenoid metabolism pathway, the mechanism of pulp softening and coloration of papaya was deduced and discussed. We illustrate that papaya fruit softening is a complex process with significant cell wall hydrolases, such as pectinases, cellulases, and hemicellulases involved in the process. Exogenous ethylene accelerates the coloration of papaya changing from green to yellow. This is likely due to the inhibition of chlorophyll biosynthesis and the α-branch of carotenoid metabolism. Chy-b may play an important role in the yellow color of papaya

  18. ETHQV6.3 is involved in melon climacteric fruit ripening and is encoded by a NAC domain transcription factor.

    PubMed

    Ríos, Pablo; Argyris, Jason; Vegas, Juan; Leida, Carmen; Kenigswald, Merav; Tzuri, Galil; Troadec, Christelle; Bendahmane, Abdelhafid; Katzir, Nurit; Picó, Belén; Monforte, Antonio J; Garcia-Mas, Jordi

    2017-08-01

    Fruit ripening is divided into climacteric and non-climacteric types depending on the presence or absence of a transient rise in respiration rate and the production of autocatalytic ethylene. Melon is ideal for the study of fruit ripening, as both climacteric and non-climacteric varieties exist. Two introgressions of the non-climacteric accession PI 161375, encompassed in the QTLs ETHQB3.5 and ETHQV6.3, into the non-climacteric 'Piel de Sapo' background are able to induce climacteric ripening independently. We report that the gene underlying ETHQV6.3 is MELO3C016540 (CmNAC-NOR), encoding a NAC (NAM, ATAF1,2, CUC2) transcription factor that is closely related to the tomato NOR (non-ripening) gene. CmNAC-NOR was functionally validated through the identification of two TILLING lines carrying non-synonymous mutations in the conserved NAC domain region. In an otherwise highly climacteric genetic background, both mutations provoked a significant delay in the onset of fruit ripening and in the biosynthesis of ethylene. The PI 161375 allele of ETHQV6.3 is similar to that of climacteric lines of the cantalupensis type and, when introgressed into the non-climacteric 'Piel de Sapo', partially restores its climacteric ripening capacity. CmNAC-NOR is expressed in fruit flesh of both climacteric and non-climacteric lines, suggesting that the causal mutation may not be acting at the transcriptional level. The use of a comparative genetic approach in a species with both climacteric and non-climacteric ripening is a powerful strategy to dissect the complex mechanisms regulating the onset of fruit ripening. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

  19. Ethylene Control of Fruit Ripening: Revisiting the Complex Network of Transcriptional Regulation1

    PubMed Central

    Chervin, Christian; Bouzayen, Mondher

    2015-01-01

    The plant hormone ethylene plays a key role in climacteric fruit ripening. Studies on components of ethylene signaling have revealed a linear transduction pathway leading to the activation of ethylene response factors. However, the means by which ethylene selects the ripening-related genes and interacts with other signaling pathways to regulate the ripening process are still to be elucidated. Using tomato (Solanum lycopersicum) as a reference species, the present review aims to revisit the mechanisms by which ethylene regulates fruit ripening by taking advantage of new tools available to perform in silico studies at the genome-wide scale, leading to a global view on the expression pattern of ethylene biosynthesis and response genes throughout ripening. Overall, it provides new insights on the transcriptional network by which this hormone coordinates the ripening process and emphasizes the interplay between ethylene and ripening-associated developmental factors and the link between epigenetic regulation and ethylene during fruit ripening. PMID:26511917

  20. The Tomato FRUITFULL Homologs TDR4/FUL1 and MBP7/FUL2 Regulate Ethylene-Independent Aspects of Fruit Ripening[W

    PubMed Central

    Bemer, Marian; Karlova, Rumyana; Ballester, Ana Rosa; Tikunov, Yury M.; Bovy, Arnaud G.; Wolters-Arts, Mieke; Rossetto, Priscilla de Barros; Angenent, Gerco C.; de Maagd, Ruud A.

    2012-01-01

    Tomato (Solanum lycopersicum) contains two close homologs of the Arabidopsis thaliana MADS domain transcription factor FRUITFULL (FUL), FUL1 (previously called TDR4) and FUL2 (previously MBP7). Both proteins interact with the ripening regulator RIPENING INHIBITOR (RIN) and are expressed during fruit ripening. To elucidate their function in tomato, we characterized single and double FUL1 and FUL2 knockdown lines. Whereas the single lines only showed very mild alterations in fruit pigmentation, the double silenced lines exhibited an orange-ripe fruit phenotype due to highly reduced lycopene levels, suggesting that FUL1 and FUL2 have a redundant function in fruit ripening. More detailed analyses of the phenotype, transcriptome, and metabolome of the fruits silenced for both FUL1 and FUL2 suggest that the genes are involved in cell wall modification, the production of cuticle components and volatiles, and glutamic acid (Glu) accumulation. Glu is responsible for the characteristic umami taste of the present-day cultivated tomato fruit. In contrast with previously identified ripening regulators, FUL1 and FUL2 do not regulate ethylene biosynthesis but influence ripening in an ethylene-independent manner. Our data combined with those of others suggest that FUL1/2 and TOMATO AGAMOUS-LIKE1 regulate different subsets of the known RIN targets, probably in a protein complex with the latter. PMID:23136376

  1. Tomato Fruit Chromoplasts Behave as Respiratory Bioenergetic Organelles during Ripening1[W][OPEN

    PubMed Central

    Renato, Marta; Pateraki, Irini; Boronat, Albert; Azcón-Bieto, Joaquín

    2014-01-01

    During tomato (Solanum lycopersicum) fruit ripening, chloroplasts differentiate into photosynthetically inactive chromoplasts. It was recently reported that tomato chromoplasts can synthesize ATP through a respiratory process called chromorespiration. Here we show that chromoplast oxygen consumption is stimulated by the electron donors NADH and NADPH and is sensitive to octyl gallate (Ogal), a plastidial terminal oxidase inhibitor. The ATP synthesis rate of isolated chromoplasts was dependent on the supply of NAD(P)H and was fully inhibited by Ogal. It was also inhibited by the proton uncoupler carbonylcyanide m-chlorophenylhydrazone, suggesting the involvement of a chemiosmotic gradient. In addition, ATP synthesis was sensitive to 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, a cytochrome b6f complex inhibitor. The possible participation of this complex in chromorespiration was supported by the detection of one of its components (cytochrome f) in chromoplasts using immunoblot and immunocytochemical techniques. The observed increased expression of cytochrome c6 during ripening suggests that it could act as electron acceptor of the cytochrome b6f complex in chromorespiration. The effects of Ogal on respiration and ATP levels were also studied in tissue samples. Oxygen uptake of mature green fruit and leaf tissues was not affected by Ogal, but was inhibited increasingly in fruit pericarp throughout ripening (up to 26% in red fruit). Similarly, Ogal caused a significant decrease in ATP content of red fruit pericarp. The number of energized mitochondria, as determined by confocal microscopy, strongly decreased in fruit tissue during ripening. Therefore, the contribution of chromoplasts to total fruit respiration appears to increase in late ripening stages. PMID:25125503

  2. Uniform ripening (U) encodes a Golden 2-like transcription factor regulating tomato fruit chloroplast development

    USDA-ARS?s Scientific Manuscript database

    Modern tomato (Solanum lycopersicum) varieties are bred for recessive uniform ripening (u) light green fruit phenotypes to facilitate maturity determinations without information about the underlying gene. We show that U encodes a Golden 2-like (GLK) transcription factor, SlGLK2, which determines the...

  3. Comparative genomics reveals candidate carotenoid pathway regulators of ripening watermelon fruit.

    PubMed

    Grassi, Stefania; Piro, Gabriella; Lee, Je Min; Zheng, Yi; Fei, Zhangjun; Dalessandro, Giuseppe; Giovannoni, James J; Lenucci, Marcello S

    2013-11-12

    Many fruits, including watermelon, are proficient in carotenoid accumulation during ripening. While most genes encoding steps in the carotenoid biosynthetic pathway have been cloned, few transcriptional regulators of these genes have been defined to date. Here we describe the identification of a set of putative carotenoid-related transcription factors resulting from fresh watermelon carotenoid and transcriptome analysis during fruit development and ripening. Our goal is to both clarify the expression profiles of carotenoid pathway genes and to identify candidate regulators and molecular targets for crop improvement. Total carotenoids progressively increased during fruit ripening up to ~55 μg g(-1) fw in red-ripe fruits. Trans-lycopene was the carotenoid that contributed most to this increase. Many of the genes related to carotenoid metabolism displayed changing expression levels during fruit ripening generating a metabolic flux toward carotenoid synthesis. Constitutive low expression of lycopene cyclase genes resulted in lycopene accumulation. RNA-seq expression profiling of watermelon fruit development yielded a set of transcription factors whose expression was correlated with ripening and carotenoid accumulation. Nineteen putative transcription factor genes from watermelon and homologous to tomato carotenoid-associated genes were identified. Among these, six were differentially expressed in the flesh of both species during fruit development and ripening. Taken together the data suggest that, while the regulation of a common set of metabolic genes likely influences carotenoid synthesis and accumulation in watermelon and tomato fruits during development and ripening, specific and limiting regulators may differ between climacteric and non-climacteric fruits, possibly related to their differential susceptibility to and use of ethylene during ripening.

  4. Polyamines Regulate Strawberry Fruit Ripening by Abscisic Acid, Auxin, and Ethylene.

    PubMed

    Guo, Jiaxuan; Wang, Shufang; Yu, Xiaoyang; Dong, Rui; Li, Yuzhong; Mei, Xurong; Shen, Yuanyue

    2018-05-01

    Polyamines (PAs) participate in many plant growth and developmental processes, including fruit ripening. However, it is not clear whether PAs play a role in the ripening of strawberry ( Fragaria ananassa ), a model nonclimacteric plant. Here, we found that the content of the PA spermine (Spm) increased more sharply after the onset of fruit coloration than did that of the PAs putrescine (Put) or spermidine (Spd). Spm dominance in ripe fruit resulted from abundant transcripts of a strawberry S -adenosyl-l-Met decarboxylase gene ( FaSAMDC ), which encodes an enzyme that generates a residue needed for PA biosynthesis. Exogenous Spm and Spd promoted fruit coloration, while exogenous Put and a SAMDC inhibitor inhibited coloration. Based on transcriptome data, up- and down-regulation of FaSAMDC expression promoted and inhibited ripening, respectively, which coincided with changes in several physiological parameters and their corresponding gene transcripts, including firmness, anthocyanin content, sugar content, polyamine content, auxin (indole-3-acetic acid [IAA]) content, abscisic acid (ABA) content, and ethylene emission. Using isothermal titration calorimetry, we found that FaSAMDC also had a high enzymatic activity with a K d of 1.7 × 10 -3 m In conclusion, PAs, especially Spm, regulate strawberry fruit ripening in an ABA-dominated, IAA-participating, and ethylene-coordinated manner, and FaSAMDC plays an important role in ripening. © 2018 American Society of Plant Biologists. All Rights Reserved.

  5. Interference with ethylene perception at receptor level sheds light on auxin and transcriptional circuits associated with the climacteric ripening of apple fruit (Malus x domestica Borkh.).

    PubMed

    Tadiello, Alice; Longhi, Sara; Moretto, Marco; Ferrarini, Alberto; Tononi, Paola; Farneti, Brian; Busatto, Nicola; Vrhovsek, Urska; Molin, Alessandra Dal; Avanzato, Carla; Biasioli, Franco; Cappellin, Luca; Scholz, Matthias; Velasco, Riccardo; Trainotti, Livio; Delledonne, Massimo; Costa, Fabrizio

    2016-12-01

    Apple (Malus x domestica Borkh.) is a model species for studying the metabolic changes that occur at the onset of ripening in fruit crops, and the physiological mechanisms that are governed by the hormone ethylene. In this study, to dissect the climacteric interplay in apple, a multidisciplinary approach was employed. To this end, a comprehensive analysis of gene expression together with the investigation of several physiological entities (texture, volatilome and content of polyphenolic compounds) was performed throughout fruit development and ripening. The transcriptomic profiling was conducted with two microarray platforms: a dedicated custom array (iRIPE) and a whole genome array specifically enriched with ripening-related genes for apple (WGAA). The transcriptomic and phenotypic changes following the application of 1-methylcyclopropene (1-MCP), an ethylene inhibitor leading to important modifications in overall fruit physiology, were also highlighted. The integrative comparative network analysis showed both negative and positive correlations between ripening-related transcripts and the accumulation of specific metabolites or texture components. The ripening distortion caused by the inhibition of ethylene perception, in addition to affecting the ethylene pathway, stimulated the de-repression of auxin-related genes, transcription factors and photosynthetic genes. Overall, the comprehensive repertoire of results obtained here advances the elucidation of the multi-layered climacteric mechanism of fruit ripening, thus suggesting a possible transcriptional circuit governed by hormones and transcription factors. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

  6. Comparative genomics reveals candidate carotenoid pathway regulators of ripening watermelon fruit

    PubMed Central

    2013-01-01

    Background Many fruits, including watermelon, are proficient in carotenoid accumulation during ripening. While most genes encoding steps in the carotenoid biosynthetic pathway have been cloned, few transcriptional regulators of these genes have been defined to date. Here we describe the identification of a set of putative carotenoid-related transcription factors resulting from fresh watermelon carotenoid and transcriptome analysis during fruit development and ripening. Our goal is to both clarify the expression profiles of carotenoid pathway genes and to identify candidate regulators and molecular targets for crop improvement. Results Total carotenoids progressively increased during fruit ripening up to ~55 μg g-1 fw in red-ripe fruits. Trans-lycopene was the carotenoid that contributed most to this increase. Many of the genes related to carotenoid metabolism displayed changing expression levels during fruit ripening generating a metabolic flux toward carotenoid synthesis. Constitutive low expression of lycopene cyclase genes resulted in lycopene accumulation. RNA-seq expression profiling of watermelon fruit development yielded a set of transcription factors whose expression was correlated with ripening and carotenoid accumulation. Nineteen putative transcription factor genes from watermelon and homologous to tomato carotenoid-associated genes were identified. Among these, six were differentially expressed in the flesh of both species during fruit development and ripening. Conclusions Taken together the data suggest that, while the regulation of a common set of metabolic genes likely influences carotenoid synthesis and accumulation in watermelon and tomato fruits during development and ripening, specific and limiting regulators may differ between climacteric and non-climacteric fruits, possibly related to their differential susceptibility to and use of ethylene during ripening. PMID:24219562

  7. The Zinc Finger Transcription Factor SlZFP2 Negatively Regulates Abscisic Acid Biosynthesis and Fruit Ripening in Tomato1

    PubMed Central

    Weng, Lin; Zhao, Fangfang; Li, Rong; Xu, Changjie; Chen, Kunsong

    2015-01-01

    Abscisic acid (ABA) regulates plant development and adaptation to environmental conditions. Although the ABA biosynthesis pathway in plants has been thoroughly elucidated, how ABA biosynthetic genes are regulated at the molecular level during plant development is less well understood. Here, we show that the tomato (Solanum lycopersicum) zinc finger transcription factor SlZFP2 is involved in the regulation of ABA biosynthesis during fruit development. Overexpression of SlZFP2 resulted in multiple phenotypic changes, including more branches, early flowering, delayed fruit ripening, lighter seeds, and faster seed germination, whereas down-regulation of its expression caused problematic fruit set, accelerated ripening, and inhibited seed germination. SlZFP2 represses ABA biosynthesis during fruit development through direct suppression of the ABA biosynthetic genes NOTABILIS, SITIENS, and FLACCA and the aldehyde oxidase SlAO1. We also show that SlZFP2 regulates fruit ripening through transcriptional suppression of the ripening regulator COLORLESS NON-RIPENING. Using bacterial one-hybrid screening and a selected amplification and binding assay, we identified the (A/T)(G/C)TT motif as the core binding sequence of SlZFP2. Furthermore, by RNA sequencing profiling, we found that 193 genes containing the SlZFP2-binding motifs in their promoters were differentially expressed in 2 d post anthesis fruits between the SlZFP2 RNA interference line and its nontransgenic sibling. We propose that SlZFP2 functions as a repressor to fine-tune ABA biosynthesis during fruit development and provides a potentially valuable tool for dissecting the role of ABA in fruit ripening. PMID:25637453

  8. Molecular characterization of banana NAC transcription factors and their interactions with ethylene signalling component EIL during fruit ripening.

    PubMed

    Shan, Wei; Kuang, Jian-fei; Chen, Lei; Xie, Hui; Peng, Huan-huan; Xiao, Yun-yi; Li, Xue-ping; Chen, Wei-xin; He, Quan-guang; Chen, Jian-ye; Lu, Wang-jin

    2012-09-01

    The plant-specific NAC (NAM, ATAF1/2, and CUC2) transcription factors (TFs) play important roles in plant growth, development, and stress responses. However, the precise role of NAC TFs in relation to fruit ripening is poorly understood. In this study, six NAC genes, designated MaNAC1-MaNAC6, were isolated and characterized from banana fruit. Subcellular localization showed that MaNAC1-MaNAC5 proteins localized preferentially to the nucleus, while MaNAC6 was distributed throughout the entire cell. A transactivation assay in yeast demonstrated that MaNAC4 and MaNAC6, as well as their C-terminal regions, possessed trans-activation activity. Gene expression profiles in fruit with four different ripening characteristics, including natural, ethylene-induced, 1-methylcyclopropene (1-MCP)-delayed, and a combination of 1-MCP with ethylene treatment, revealed that the MaNAC genes were differentially expressed in peel and pulp during post-harvest ripening. MaNAC1 and MaNAC2 were apparently upregulated by ethylene in peel and pulp, consistent with the increase in ethylene production. In contrast, MaNAC3 in peel and pulp and MaNAC5 in peel were constitutively expressed, and transcripts of MaNAC4 in peel and pulp and MaNAC6 in peel decreased, while MaNAC5 or MaNAC6 in pulp increased slightly during fruit ripening. Furthermore, the MaNAC2 promoter was activated after ethylene application, further enhancing the involvement of MaNAC2 in fruit ripening. More importantly, yeast two-hybrid and bimolecular fluorescence complementation analyses confirmed that MaNAC1/2 physically interacted with a downstream component of ethylene signalling, ethylene insensitive 3 (EIN3)-like protein, termed MaEIL5, which was downregulated during ripening. Taken together, these results suggest that MaNACs such as MaNAC1/MaNAC2, may be involved in banana fruit ripening via interaction with ethylene signalling components.

  9. Molecular characterization of banana NAC transcription factors and their interactions with ethylene signalling component EIL during fruit ripening

    PubMed Central

    Shan, Wei; Kuang, Jian-fei; Chen, Jian-ye; Lu, Wang-jin

    2012-01-01

    The plant-specific NAC (NAM, ATAF1/2, and CUC2) transcription factors (TFs) play important roles in plant growth, development, and stress responses. However, the precise role of NAC TFs in relation to fruit ripening is poorly understood. In this study, six NAC genes, designated MaNAC1–MaNAC6, were isolated and characterized from banana fruit. Subcellular localization showed that MaNAC1–MaNAC5 proteins localized preferentially to the nucleus, while MaNAC6 was distributed throughout the entire cell. A transactivation assay in yeast demonstrated that MaNAC4 and MaNAC6, as well as their C-terminal regions, possessed trans-activation activity. Gene expression profiles in fruit with four different ripening characteristics, including natural, ethylene-induced, 1-methylcyclopropene (1-MCP)-delayed, and a combination of 1-MCP with ethylene treatment, revealed that the MaNAC genes were differentially expressed in peel and pulp during post-harvest ripening. MaNAC1 and MaNAC2 were apparently upregulated by ethylene in peel and pulp, consistent with the increase in ethylene production. In contrast, MaNAC3 in peel and pulp and MaNAC5 in peel were constitutively expressed, and transcripts of MaNAC4 in peel and pulp and MaNAC6 in peel decreased, while MaNAC5 or MaNAC6 in pulp increased slightly during fruit ripening. Furthermore, the MaNAC2 promoter was activated after ethylene application, further enhancing the involvement of MaNAC2 in fruit ripening. More importantly, yeast two-hybrid and bimolecular fluorescence complementation analyses confirmed that MaNAC1/2 physically interacted with a downstream component of ethylene signalling, ethylene insensitive 3 (EIN3)-like protein, termed MaEIL5, which was downregulated during ripening. Taken together, these results suggest that MaNACs such as MaNAC1/MaNAC2, may be involved in banana fruit ripening via interaction with ethylene signalling components. PMID:22888129

  10. The transcriptional regulatory network mediated by banana (Musa acuminata) dehydration-responsive element binding (MaDREB) transcription factors in fruit ripening.

    PubMed

    Kuang, Jian-Fei; Chen, Jian-Ye; Liu, Xun-Cheng; Han, Yan-Chao; Xiao, Yun-Yi; Shan, Wei; Tang, Yang; Wu, Ke-Qiang; He, Jun-Xian; Lu, Wang-Jin

    2017-04-01

    Fruit ripening is a complex, genetically programmed process involving the action of critical transcription factors (TFs). Despite the established significance of dehydration-responsive element binding (DREB) TFs in plant abiotic stress responses, the involvement of DREBs in fruit ripening is yet to be determined. Here, we identified four genes encoding ripening-regulated DREB TFs in banana (Musa acuminata), MaDREB1, MaDREB2, MaDREB3, and MaDREB4, and demonstrated that they play regulatory roles in fruit ripening. We showed that MaDREB1-MaDREB4 are nucleus-localized, induced by ethylene and encompass transcriptional activation activities. We performed a genome-wide chromatin immunoprecipitation and high-throughput sequencing (ChIP-Seq) experiment for MaDREB2 and identified 697 genomic regions as potential targets of MaDREB2. MaDREB2 binds to hundreds of loci with diverse functions and its binding sites are distributed in the promoter regions proximal to the transcriptional start site (TSS). Most of the MaDREB2-binding targets contain the conserved (A/G)CC(G/C)AC motif and MaDREB2 appears to directly regulate the expression of a number of genes involved in fruit ripening. In combination with transcriptome profiling (RNA sequencing) data, our results indicate that MaDREB2 may serve as both transcriptional activator and repressor during banana fruit ripening. In conclusion, our study suggests a hierarchical regulatory model of fruit ripening in banana and that the MaDREB TFs may act as transcriptional regulators in the regulatory network. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  11. Ethylene biosynthesis and perception during ripening of loquat fruit (Eriobotrya japonica Lindl.).

    PubMed

    Alos, E; Martinez-Fuentes, A; Reig, C; Mesejo, C; Rodrigo, M J; Agustí, M; Zacarías, L

    2017-03-01

    In order to gain insights into the controversial ripening behavior of loquat fruits, in the present study we have analyzed the expression of three genes related to ethylene biosynthesis (ACS1, ACO1 and ACO2), two ethylene receptors (ERS1a and ERS1b), one signal transduction component (CTR1) and one transcription factor (EIL1) in peel and pulp of loquat fruit during natural ripening and also in fruits treated with ethylene (10μLL -1 ) and 1-MCP (10μLL -1 ), an ethylene action inhibitor. In fruits attached to or detached from the tree, a slight increase in ethylene production was detected at the yellow stage, but the respiration rate declined progressively during ripening. Accumulation of transcripts of ethylene biosynthetic genes did not correlate with changes in ethylene production, since the maximum accumulation of ACS1 and ACO1 mRNA was detected in fully coloured fruits. Expression of ethylene receptor and signaling genes followed a different pattern in peel and pulp tissues. After fruit detachment and incubation at 20°C for up to 6days, ACS1 mRNA slightly increased, ACO1 experienced a substantial increment and ACO2 declined. In the peel, these changes were advanced by exogenous ethylene and partially inhibited by 1-MCP. In the pulp, 1-MCP repressed most of the changes in the expression of biosynthetic genes, while ethylene had almost no effects. Expression of ethylene perception and signaling genes was barely affected by ethylene or 1-MCP. Collectively, a differential transcriptional regulation of ethylene biosynthetic genes operates in peel and pulp, and support the notion of non-climacteric ripening in loquat fruits. Ethylene action, however, appears to be required to sustain or maintain the expression of specific genes. Copyright © 2016. Published by Elsevier GmbH.

  12. A DEMETER-like DNA demethylase governs tomato fruit ripening.

    PubMed

    Liu, Ruie; How-Kit, Alexandre; Stammitti, Linda; Teyssier, Emeline; Rolin, Dominique; Mortain-Bertrand, Anne; Halle, Stefanie; Liu, Mingchun; Kong, Junhua; Wu, Chaoqun; Degraeve-Guibault, Charlotte; Chapman, Natalie H; Maucourt, Mickael; Hodgman, T Charlie; Tost, Jörg; Bouzayen, Mondher; Hong, Yiguo; Seymour, Graham B; Giovannoni, James J; Gallusci, Philippe

    2015-08-25

    In plants, genomic DNA methylation which contributes to development and stress responses can be actively removed by DEMETER-like DNA demethylases (DMLs). Indeed, in Arabidopsis DMLs are important for maternal imprinting and endosperm demethylation, but only a few studies demonstrate the developmental roles of active DNA demethylation conclusively in this plant. Here, we show a direct cause and effect relationship between active DNA demethylation mainly mediated by the tomato DML, SlDML2, and fruit ripening- an important developmental process unique to plants. RNAi SlDML2 knockdown results in ripening inhibition via hypermethylation and repression of the expression of genes encoding ripening transcription factors and rate-limiting enzymes of key biochemical processes such as carotenoid synthesis. Our data demonstrate that active DNA demethylation is central to the control of ripening in tomato.

  13. The banana fruit Dof transcription factor MaDof23 acts as a repressor and interacts with MaERF9 in regulating ripening-related genes

    PubMed Central

    Feng, Bi-hong; Han, Yan-chao; Xiao, Yun-yi; Kuang, Jian-fei; Fan, Zhong-qi; Chen, Jian-ye; Lu, Wang-jin

    2016-01-01

    The DNA binding with one finger (Dof) proteins, a family of plant-specific transcription factors, are involved in a variety of plant biological processes. However, little information is available on their involvement in fruit ripening. We have characterized 25 MaDof genes from banana fruit (Musa acuminata), designated as MaDof1–MaDof25. Gene expression analysis in fruit subjected to different ripening conditions revealed that MaDofs were differentially expressed during different stages of ripening. MaDof10, 23, 24, and 25 were ethylene-inducible and nuclear-localized, and their transcript levels increased during fruit ripening. Moreover, yeast two-hybrid and bimolecular fluorescence complementation analyses demonstrated a physical interaction between MaDof23 and MaERF9, a potential regulator of fruit ripening reported in a previous study. We determined that MaDof23 is a transcriptional repressor, whereas MaERF9 is a transcriptional activator. We suggest that they might act antagonistically in regulating 10 ripening-related genes, including MaEXP1/2/3/5, MaXET7, MaPG1, MaPME3, MaPL2, MaCAT, and MaPDC, which are associated with cell wall degradation and aroma formation. Taken together, our findings provide new insight into the transcriptional regulation network controlling banana fruit ripening. PMID:26889012

  14. Peptides interfering with protein-protein interactions in the ethylene signaling pathway delay tomato fruit ripening

    NASA Astrophysics Data System (ADS)

    Bisson, Melanie M. A.; Kessenbrock, Mareike; Müller, Lena; Hofmann, Alexander; Schmitz, Florian; Cristescu, Simona M.; Groth, Georg

    2016-08-01

    The plant hormone ethylene is involved in the regulation of several processes with high importance for agricultural applications, e.g. ripening, aging and senescence. Previous work in our group has identified a small peptide (NOP-1) derived from the nuclear localization signal of the Arabidopsis ethylene regulator ETHYLENE INSENSITIVE-2 (EIN2) C-terminal part as efficient inhibitor of ethylene responses. Here, we show that NOP-1 is also able to efficiently disrupt EIN2-ETR1 complex formation in tomato, indicating that the NOP-1 inhibition mode is conserved across plant species. Surface application of NOP-1 on green tomato fruits delays ripening similar to known inhibitors of ethylene perception (MCP) and ethylene biosynthesis (AVG). Fruits treated with NOP-1 showed similar ethylene production as untreated controls underlining that NOP-1 blocks ethylene signaling by targeting an essential interaction in this pathway, while having no effect on ethylene biosynthesis.

  15. Peptides interfering with protein-protein interactions in the ethylene signaling pathway delay tomato fruit ripening.

    PubMed

    Bisson, Melanie M A; Kessenbrock, Mareike; Müller, Lena; Hofmann, Alexander; Schmitz, Florian; Cristescu, Simona M; Groth, Georg

    2016-08-01

    The plant hormone ethylene is involved in the regulation of several processes with high importance for agricultural applications, e.g. ripening, aging and senescence. Previous work in our group has identified a small peptide (NOP-1) derived from the nuclear localization signal of the Arabidopsis ethylene regulator ETHYLENE INSENSITIVE-2 (EIN2) C-terminal part as efficient inhibitor of ethylene responses. Here, we show that NOP-1 is also able to efficiently disrupt EIN2-ETR1 complex formation in tomato, indicating that the NOP-1 inhibition mode is conserved across plant species. Surface application of NOP-1 on green tomato fruits delays ripening similar to known inhibitors of ethylene perception (MCP) and ethylene biosynthesis (AVG). Fruits treated with NOP-1 showed similar ethylene production as untreated controls underlining that NOP-1 blocks ethylene signaling by targeting an essential interaction in this pathway, while having no effect on ethylene biosynthesis.

  16. Involvement of a banana MADS-box transcription factor gene in ethylene-induced fruit ripening.

    PubMed

    Liu, Juhua; Xu, Biyu; Hu, Lifang; Li, Meiying; Su, Wei; Wu, Jing; Yang, Jinghao; Jin, Zhiqiang

    2009-01-01

    To investigate the regulation of MADS-box genes in banana (Musa acuminata L. AAA group cv. Brazilian) fruit development and postharvest ripening, we isolated from banana fruit a MADS-box gene designated MuMADS1. Amino acid alignment indicated MuMADS1 belongs to the AGAMOUS subfamily, and phylogenetic analysis indicates that this gene is most similar to class D MADS-box genes. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that MuMADS1 is expressed in the stamen and pistil of male and female flowers and in the rhizome, the vegetative reproductive organ of the banana plant. In preharvest banana fruit, MuMADS1 is likely expressed throughout banana fruit development. In postharvest banana ripening, MuMADS1 is associated with ethylene biosynthesis. Expression patterns of MuMADS1 during postharvest ripening as determined by real-time RT-PCR suggest that differential expression of MuMADS1 may not only be induced by ethylene biosynthesis associated with postharvest banana ripening, but also may be induced by exogenous ethylene.

  17. Ripening Behavior of Wild Tomato Species 1

    PubMed Central

    Grumet, Rebecca; Fobes, Jon F.; Herner, Robert C.

    1981-01-01

    Nine wild tomato species were surveyed for variability in ripening characteristics. External signs of ripening, age of fruit at ripening, and ethylene production patterns were compared. Ethylene production was monitored using an ethylene-free air stream system and gas chromatography. Based on these ripening characteristics, the fruits fell into three general categories: those that change color when they ripen, green-fruited species that abscise prior to ripening, and green-fruited species that ripen on the vine. The fruits that change color, Lycopersicon esculentum var. cerasiforme, Lycopersicon pimpinellifolium and Lycopersicon cheesmanii, exhibited a peak of ethylene production similar to the cultivated tomato; there were differences, however, in the timing and magnitude of the ethylene production. Peak levels of ethylene production are correlated with age at maturity. For the two species that abscise prior to ripening, Lycopersicon chilense and Lycopersicon peruvianum, ability to produce ethylene varied with stage of maturity. The two species differed from each other in time of endogenous ethylene production relative to abscission, suggesting differences in the control mechanisms regulating their ripening. For two of the green-fruited species that ripen on the vine, Lycopersicon chmielewskii and Lycopersicon parviflorum, ethylene production was correlated to fruit softening. For Lycopersicon hirsutum and Solanum pennellii, however, ethylene production was not correlated with external ripening changes, making questionable the role of ethylene as the ripening hormone in these fruits. PMID:16662121

  18. A new tomato NAC (NAM/ATAF1/2/CUC2) transcription factor, SlNAC4, functions as a positive regulator of fruit ripening and carotenoid accumulation.

    PubMed

    Zhu, Mingku; Chen, Guoping; Zhou, Shuang; Tu, Yun; Wang, Yi; Dong, Tingting; Hu, Zongli

    2014-01-01

    Fruit ripening in tomato (Solanum lycopersicum) is a complicated development process affected by both endogenous hormonal and genetic regulators and external signals. Although the role of NOR, a member of the NAC domain family, in mediating tomato fruit ripening has been established, its underlying molecular mechanisms remain unclear. To explore further the role of NAC transcription factors in fruit ripening, we characterized a new tomato NAC domain protein, named SlNAC4, which shows high accumulation in sepal and at the onset of fruit ripening. Various stress treatments including wounding, NaCl, dehydration and low temperature significantly increased the expression of SlNAC4. Reduced expression of SlNAC4 by RNA interference (RNAi) in tomato resulted in delayed fruit ripening, suppressed Chl breakdown and decreased ethylene synthesis mediated mainly through reduced expression of ethylene biosynthesis genes of system-2, and reduced carotenoids by alteration of the carotenoid pathway flux. Transgenic tomato fruits also displayed significant down-regulation of multiple ripening-associated genes, indicating that SlNAC4 functions as a positive regulator of fruit ripening by affecting ethylene synthesis and carotenoid accumulation. Moreover, we also noted that SlNAC4 could not be induced by ethylene and may function upstream of the ripening regulator RIN and positively regulate its expression. Yeast two-hybrid assay further revealed that SlNAC4 could interact with both RIN and NOR protein. These results suggested that ethylene-dependent and -independent processes are regulated by SlNAC4 in the fruit ripening regulatory network.

  19. Hypoxia and PGE2 Regulate MiTF-CX During Cervical Ripening

    PubMed Central

    Hari Kishore, Annavarapu; Li, Xiang-Hong

    2012-01-01

    The mechanisms by which the cervix remains closed during the massive uterine expansion of pregnancy are unknown. IL-8 is important for recruitment of immune cells into the cervical stroma, matrix remodeling, and dilation of the cervix during labor. Previously, we have shown that several cytokine genes transcriptionally repressed in the cervix during gestation are activated during cervical ripening and dilation. IL-8 gene expression is repressed in cervical stromal cells during pregnancy by the transcription factor microphthalmia-associated transcription factor (MiTF-CX). Here, we tested the hypothesis that hypoxia and the transcription factor hypoxia inducible factor-1α (HIF-1α) may regulate MiTF-CX and cervical ripening. Using tissues from women during pregnancy before and after cervical ripening, we show that, during cervical ripening, HIF-1α was stabilized and relocalized to the nucleus. Further, we found that hypoxia and two hypoxia mimetics that stabilize HIF-1α activated the transcriptional repressor differentiated embryo chondrocyte-expressed gene 1, which bound to sites in the MiTF-CX promoter crucial for its positive autoregulation. Ectopic overexpression of MiTF-CX abrogated hypoxia-induced up-regulation of IL-8 gene expression. We also show that activation of HIF-1α induced cyclooxygenase-2 and that prostaglandin E2 repressed MiTF-CX. We conclude that hypoxia and stabilization of the transcription factor HIF-1α result in up-regulation of differentiated embryo chondrocyte-expressed gene 1, loss of MiTF, and absence of MiTF binding to the IL-8 promoter, which in turn leads to up-regulation of IL-8 gene expression. Hypoxia also up-regulated cyclooxygenase-2, leading to prostaglandin E2-mediated loss of MiTF in cervical stromal cells. The results support a pivotal role for hypoxia and HIF-1α in the cervical ripening process during pregnancy. PMID:23144021

  20. Transcriptome Analysis of Cell Wall and NAC Domain Transcription Factor Genes during Elaeis guineensis Fruit Ripening: Evidence for Widespread Conservation within Monocot and Eudicot Lineages

    PubMed Central

    Tranbarger, Timothy J.; Fooyontphanich, Kim; Roongsattham, Peerapat; Pizot, Maxime; Collin, Myriam; Jantasuriyarat, Chatchawan; Suraninpong, Potjamarn; Tragoonrung, Somvong; Dussert, Stéphane; Verdeil, Jean-Luc; Morcillo, Fabienne

    2017-01-01

    The oil palm (Elaeis guineensis), a monocotyledonous species in the family Arecaceae, has an extraordinarily oil rich fleshy mesocarp, and presents an original model to examine the ripening processes and regulation in this particular monocot fruit. Histochemical analysis and cell parameter measurements revealed cell wall and middle lamella expansion and degradation during ripening and in response to ethylene. Cell wall related transcript profiles suggest a transition from synthesis to degradation is under transcriptional control during ripening, in particular a switch from cellulose, hemicellulose, and pectin synthesis to hydrolysis and degradation. The data provide evidence for the transcriptional activation of expansin, polygalacturonase, mannosidase, beta-galactosidase, and xyloglucan endotransglucosylase/hydrolase proteins in the ripening oil palm mesocarp, suggesting widespread conservation of these activities during ripening for monocotyledonous and eudicotyledonous fruit types. Profiling of the most abundant oil palm polygalacturonase (EgPG4) and 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) transcripts during development and in response to ethylene demonstrated both are sensitive markers of ethylene production and inducible gene expression during mesocarp ripening, and provide evidence for a conserved regulatory module between ethylene and cell wall pectin degradation. A comprehensive analysis of NAC transcription factors confirmed at least 10 transcripts from diverse NAC domain clades are expressed in the mesocarp during ripening, four of which are induced by ethylene treatment, with the two most inducible (EgNAC6 and EgNAC7) phylogenetically similar to the tomato NAC-NOR master-ripening regulator. Overall, the results provide evidence that despite the phylogenetic distance of the oil palm within the family Arecaceae from the most extensively studied monocot banana fruit, it appears ripening of divergent monocot and eudicot fruit lineages are

  1. Transcriptional Regulation of Fruit Ripening by Tomato FRUITFULL Homologs and Associated MADS Box Proteins[W

    PubMed Central

    Fujisawa, Masaki; Shima, Yoko; Nakagawa, Hiroyuki; Kitagawa, Mamiko; Kimbara, Junji; Nakano, Toshitsugu; Kasumi, Takafumi; Ito, Yasuhiro

    2014-01-01

    The tomato (Solanum lycopersicum) MADS box FRUITFULL homologs FUL1 and FUL2 act as key ripening regulators and interact with the master regulator MADS box protein RIPENING INHIBITOR (RIN). Here, we report the large-scale identification of direct targets of FUL1 and FUL2 by transcriptome analysis of FUL1/FUL2 suppressed fruits and chromatin immunoprecipitation coupled with microarray analysis (ChIP-chip) targeting tomato gene promoters. The ChIP-chip and transcriptome analysis identified FUL1/FUL2 target genes that contain at least one genomic region bound by FUL1 or FUL2 (regions that occur mainly in their promoters) and exhibit FUL1/FUL2-dependent expression during ripening. These analyses identified 860 direct FUL1 targets and 878 direct FUL2 targets; this set of genes includes both direct targets of RIN and nontargets of RIN. Functional classification of the FUL1/FUL2 targets revealed that these FUL homologs function in many biological processes via the regulation of ripening-related gene expression, both in cooperation with and independent of RIN. Our in vitro assay showed that the FUL homologs, RIN, and tomato AGAMOUS-LIKE1 form DNA binding complexes, suggesting that tetramer complexes of these MADS box proteins are mainly responsible for the regulation of ripening. PMID:24415769

  2. Ripening of Semiconductor Nanoplatelets.

    PubMed

    Ott, Florian D; Riedinger, Andreas; Ochsenbein, David R; Knüsel, Philippe N; Erwin, Steven C; Mazzotti, Marco; Norris, David J

    2017-11-08

    Ostwald ripening describes how the size distribution of colloidal particles evolves with time due to thermodynamic driving forces. Typically, small particles shrink and provide material to larger particles, which leads to size defocusing. Semiconductor nanoplatelets, thin quasi-two-dimensional (2D) particles with thicknesses of only a few atomic layers but larger lateral dimensions, offer a unique system to investigate this phenomenon. Experiments show that the distribution of nanoplatelet thicknesses does not defocus during ripening, but instead jumps sequentially from m to (m + 1) monolayers, allowing precise thickness control. We investigate how this counterintuitive process occurs in CdSe nanoplatelets. We develop a microscopic model that treats the kinetics and thermodynamics of attachment and detachment of monomers as a function of their concentration. We then simulate the growth process from nucleation through ripening. For a given thickness, we observe Ostwald ripening in the lateral direction, but none perpendicular. Thicker populations arise instead from nuclei that capture material from thinner nanoplatelets as they dissolve laterally. Optical experiments that attempt to track the thickness and lateral extent of nanoplatelets during ripening appear consistent with these conclusions. Understanding such effects can lead to better synthetic control, enabling further exploration of quasi-2D nanomaterials.

  3. A DEMETER-like DNA demethylase governs tomato fruit ripening

    PubMed Central

    Liu, Ruie; How-Kit, Alexandre; Stammitti, Linda; Teyssier, Emeline; Rolin, Dominique; Mortain-Bertrand, Anne; Halle, Stefanie; Liu, Mingchun; Kong, Junhua; Wu, Chaoqun; Degraeve-Guibault, Charlotte; Chapman, Natalie H.; Maucourt, Mickael; Hodgman, T. Charlie; Tost, Jörg; Bouzayen, Mondher; Hong, Yiguo; Seymour, Graham B.; Giovannoni, James J.; Gallusci, Philippe

    2015-01-01

    In plants, genomic DNA methylation which contributes to development and stress responses can be actively removed by DEMETER-like DNA demethylases (DMLs). Indeed, in Arabidopsis DMLs are important for maternal imprinting and endosperm demethylation, but only a few studies demonstrate the developmental roles of active DNA demethylation conclusively in this plant. Here, we show a direct cause and effect relationship between active DNA demethylation mainly mediated by the tomato DML, SlDML2, and fruit ripening— an important developmental process unique to plants. RNAi SlDML2 knockdown results in ripening inhibition via hypermethylation and repression of the expression of genes encoding ripening transcription factors and rate-limiting enzymes of key biochemical processes such as carotenoid synthesis. Our data demonstrate that active DNA demethylation is central to the control of ripening in tomato. PMID:26261318

  4. Physical, chemical and sensory characteristics of red guava (Psidium guajava) velva at different fruit ripening time

    NASA Astrophysics Data System (ADS)

    Ishartani, D.; Rahman, F. L. F.; Hartanto, R.; Utami, R.; Khasanah, L. U.

    2018-01-01

    This study purposed to determine the effect of red guava fruit ripening time on the physical (overrun and melting rate), chemical (vitamin C, pH, total dissolved solid) and sensory (color, taste, aroma, texture, and overall compare to control (without ripening) velva) characteristic of red guava velva. Red guava fruits were harvested at 90 days after flowering, ripened and then processed into velva. This research used Completely Randomized Design with fruit ripening time (without ripening, 4 days, and 6 days) as single factor. The research was conducted in triplicate. Chemical and physical characteristic data was analysed using One Way Analysis of Varian whether sensory characteristic data was analyzed using Independent Sample T-test. The result showed that fruit ripening time significantly affected the physical, chemical and sensory characteristic of the velva. Vitamin C, pH, and total solid of the velva were increased as the ripening time prolonged. In other hand, increasing of fruit ripening time decreased the overrun and melting rate of the velva. Red guava velva made from 6 days ripening had better sensory characteristics compared to velva made from red guava fruit without ripening or 4 day ripening. This research conclude that 6 days ripening time gives better chemical, physical and sensory characteristics of the velva compare to 4 days ripening time. Red guava fruits ripened for 6 days were recommended as raw material in velva making.

  5. Expression profiling of various genes during the fruit development and ripening of mango.

    PubMed

    Pandit, Sagar S; Kulkarni, Ram S; Giri, Ashok P; Köllner, Tobias G; Degenhardt, Jörg; Gershenzon, Jonathan; Gupta, Vidya S

    2010-06-01

    Mango (Mangifera indica L. cv. Alphonso) development and ripening are the programmed processes; conventional indices and volatile markers help to determine agronomically important stages of fruit life (fruit-setting, harvesting maturity and ripening climacteric). However, more and precise markers are required to understand this programming; apparently, fruit's transcriptome can be a good source of such markers. Therefore, we isolated 18 genes related to the physiology and biochemistry of the fruit and profiled their expression in developing and ripening fruits, flowers and leaves of mango using relative quantitation PCR. In most of the tissues, genes related to primary metabolism, abiotic stress, ethylene response and protein turnover showed high expression as compared to that of the genes related to flavor production. Metallothionin and/or ethylene-response transcription factor showed highest level of transcript abundance in all the tissues. Expressions of mono- and sesquiterpene synthases and 14-3-3 lowered during ripening; whereas, that of lipoxygenase, ethylene-response factor and ubiquitin-protein ligase increased during ripening. Based on these expression profiles, flower showed better positive correlation with developing and ripening fruits than leaf. Most of the genes showed their least expression on the second day of harvest, suggesting that harvesting signals significantly affect the fruit metabolism. Important stages in the fruit life were clearly indicated by the significant changes in the expression levels of various genes. These indications complemented those from the previous analyses of fruit development, ripening and volatile emission, revealing the harmony between physiological, biochemical and molecular activities of the fruit.

  6. Banana MaMADS Transcription Factors Are Necessary for Fruit Ripening and Molecular Tools to Promote Shelf-Life and Food Security.

    PubMed

    Elitzur, Tomer; Yakir, Esther; Quansah, Lydia; Zhangjun, Fei; Vrebalov, Julia; Khayat, Eli; Giovannoni, James J; Friedman, Haya

    2016-05-01

    Genetic solutions to postharvest crop loss can reduce cost and energy inputs while increasing food security, especially for banana (Musa acuminata), which is a significant component of worldwide food commerce. We have functionally characterized two banana E class (SEPALLATA3 [SEP3]) MADS box genes, MaMADS1 and MaMADS2, homologous to the tomato (Solanum lycopersicum) RIN-MADS ripening gene. Transgenic banana plants repressing either gene (via antisense or RNA interference [RNAi]) were created and exhibited specific ripening delay and extended shelf-life phenotypes, including delayed color development and softening. The delay in fruit ripening is associated with a delay in climacteric respiration and reduced synthesis of the ripening hormone ethylene; in the most severe repressed lines, no ethylene was produced and ripening was most delayed. Unlike tomato rin mutants, banana fruits of all transgenic repression lines responded to exogenous ethylene by ripening normally, likely due to incomplete transgene repression and/or compensation by other MADS box genes. Our results show that, although MADS box ripening gene necessity is conserved across diverse taxa (monocots to dicots), unlike tomato, banana ripening requires at least two necessary members of the SEPALLATA MADS box gene group, and either can serve as a target for ripening control. The utility of such genes as tools for ripening control is especially relevant in important parthenocarpic crops such as the vegetatively propagated and widely consumed Cavendish banana, where breeding options for trait improvement are severely limited. © 2016 American Society of Plant Biologists. All Rights Reserved.

  7. Interactions between ethylene and auxin are crucial to the control of grape (Vitis vinifera L.) berry ripening.

    PubMed

    Böttcher, Christine; Burbidge, Crista A; Boss, Paul K; Davies, Christopher

    2013-12-23

    Fruit development is controlled by plant hormones, but the role of hormone interactions during fruit ripening is poorly understood. Interactions between ethylene and the auxin indole-3-acetic acid (IAA) are likely to be crucial during the ripening process, since both hormones have been shown to be implicated in the control of ripening in a range of different fruit species. Grapevine (Vitis vinifera L.) homologues of the TRYPTOPHAN AMINOTRANSFERASE RELATED (TAR) and YUCCA families, functioning in the only characterized pathway of auxin biosynthesis, were identified and the expression of several TAR genes was shown to be induced by the pre-ripening application of the ethylene-releasing compound Ethrel. The induction of TAR expression was accompanied by increased IAA and IAA-Asp concentrations, indicative of an upregulation of auxin biosynthesis and conjugation. Exposure of ex planta, pre-ripening berries to the ethylene biosynthesis inhibitor aminoethoxyvinylglycine resulted in decreased IAA and IAA-Asp concentrations. The delayed initiation of ripening observed in Ethrel-treated berries might therefore represent an indirect ethylene effect mediated by increased auxin concentrations. During berry development, the expression of three TAR genes and one YUCCA gene was upregulated at the time of ripening initiation and/or during ripening. This increase in auxin biosynthesis gene expression was preceded by high expression levels of the ethylene biosynthesis genes 1-aminocyclopropane-1-carboxylate synthase and 1-aminocyclopropane-1-carboxylate oxidase. In grape berries, members of both gene families involved in the two-step pathway of auxin biosynthesis are expressed, suggesting that IAA is produced through the combined action of TAR and YUCCA proteins in developing berries. The induction of TAR expression by Ethrel applications and the developmental expression patterns of auxin and ethylene biosynthesis genes indicate that elevated concentrations of ethylene prior to the

  8. Interactions between ethylene and auxin are crucial to the control of grape (Vitis vinifera L.) berry ripening

    PubMed Central

    2013-01-01

    Background Fruit development is controlled by plant hormones, but the role of hormone interactions during fruit ripening is poorly understood. Interactions between ethylene and the auxin indole-3-acetic acid (IAA) are likely to be crucial during the ripening process, since both hormones have been shown to be implicated in the control of ripening in a range of different fruit species. Results Grapevine (Vitis vinifera L.) homologues of the TRYPTOPHAN AMINOTRANSFERASE RELATED (TAR) and YUCCA families, functioning in the only characterized pathway of auxin biosynthesis, were identified and the expression of several TAR genes was shown to be induced by the pre-ripening application of the ethylene-releasing compound Ethrel. The induction of TAR expression was accompanied by increased IAA and IAA-Asp concentrations, indicative of an upregulation of auxin biosynthesis and conjugation. Exposure of ex planta, pre-ripening berries to the ethylene biosynthesis inhibitor aminoethoxyvinylglycine resulted in decreased IAA and IAA-Asp concentrations. The delayed initiation of ripening observed in Ethrel-treated berries might therefore represent an indirect ethylene effect mediated by increased auxin concentrations. During berry development, the expression of three TAR genes and one YUCCA gene was upregulated at the time of ripening initiation and/or during ripening. This increase in auxin biosynthesis gene expression was preceded by high expression levels of the ethylene biosynthesis genes 1-aminocyclopropane-1-carboxylate synthase and 1-aminocyclopropane-1-carboxylate oxidase. Conclusions In grape berries, members of both gene families involved in the two-step pathway of auxin biosynthesis are expressed, suggesting that IAA is produced through the combined action of TAR and YUCCA proteins in developing berries. The induction of TAR expression by Ethrel applications and the developmental expression patterns of auxin and ethylene biosynthesis genes indicate that elevated

  9. The Basic Leucine Zipper Transcription Factor ABSCISIC ACID RESPONSE ELEMENT-BINDING FACTOR2 Is an Important Transcriptional Regulator of Abscisic Acid-Dependent Grape Berry Ripening Processes1[W][OPEN

    PubMed Central

    Nicolas, Philippe; Lecourieux, David; Kappel, Christian; Cluzet, Stéphanie; Cramer, Grant; Delrot, Serge; Lecourieux, Fatma

    2014-01-01

    In grape (Vitis vinifera), abscisic acid (ABA) accumulates during fruit ripening and is thought to play a pivotal role in this process, but the molecular basis of this control is poorly understood. This work characterizes ABSCISIC ACID RESPONSE ELEMENT-BINDING FACTOR2 (VvABF2), a grape basic leucine zipper transcription factor belonging to a phylogenetic subgroup previously shown to be involved in ABA and abiotic stress signaling in other plant species. VvABF2 transcripts mainly accumulated in the berry, from the onset of ripening to the harvesting stage, and were up-regulated by ABA. Microarray analysis of transgenic grape cells overexpressing VvABF2 showed that this transcription factor up-regulates and/or modifies existing networks related to ABA responses. In addition, grape cells overexpressing VvABF2 exhibited enhanced responses to ABA treatment compared with control cells. Among the VvABF2-mediated responses highlighted in this study, the synthesis of phenolic compounds and cell wall softening were the most strongly affected. VvABF2 overexpression strongly increased the accumulation of stilbenes that play a role in plant defense and human health (resveratrol and piceid). In addition, the firmness of fruits from tomato (Solanum lycopersicum) plants overexpressing VvABF2 was strongly reduced. These data indicate that VvABF2 is an important transcriptional regulator of ABA-dependent grape berry ripening. PMID:24276949

  10. Cold shock treatment extends shelf life of naturally ripened or ethylene-ripened avocado fruits.

    PubMed

    Chen, Jiao; Liu, Xixia; Li, Fenfang; Li, Yixing; Yuan, Debao

    2017-01-01

    Avocado is an important tropical fruit with high commercial value, but has a relatively short storage life. In this study, the effects of cold shock treatment (CST) on shelf life of naturally ripened and ethylene-ripened avocado fruits were investigated. Fruits were immersed in ice water for 30 min, then subjected to natural or ethylene-induced ripening. Fruit color; firmness; respiration rate; ethylene production; and the activities of polygalacturonase (PG), pectin methylesterase (PME), and endo-β-1,4-glucanase were measured. Immersion in ice water for 30 min effectively delayed ripening-associated processes, including peel discoloration, pulp softening, respiration rate, and ethylene production during shelf life. The delay in fruit softening by CST was associated with decreased PG and endo-β-1,4-glucanase activities, but not PME activity. This method could potentially be a useful postharvest technology to extend shelf life of avocado fruits.

  11. Cold shock treatment extends shelf life of naturally ripened or ethylene-ripened avocado fruits

    PubMed Central

    Li, Fenfang; Li, Yixing

    2017-01-01

    Avocado is an important tropical fruit with high commercial value, but has a relatively short storage life. In this study, the effects of cold shock treatment (CST) on shelf life of naturally ripened and ethylene-ripened avocado fruits were investigated. Fruits were immersed in ice water for 30 min, then subjected to natural or ethylene-induced ripening. Fruit color; firmness; respiration rate; ethylene production; and the activities of polygalacturonase (PG), pectin methylesterase (PME), and endo-β-1,4-glucanase were measured. Immersion in ice water for 30 min effectively delayed ripening-associated processes, including peel discoloration, pulp softening, respiration rate, and ethylene production during shelf life. The delay in fruit softening by CST was associated with decreased PG and endo-β-1,4-glucanase activities, but not PME activity. This method could potentially be a useful postharvest technology to extend shelf life of avocado fruits. PMID:29253879

  12. Irradiated Chinese Rugao ham: Changes in volatile N-nitrosamine, biogenic amine and residual nitrite during ripening and post-ripening.

    PubMed

    Wei, Fashan; Xu, Xinglian; Zhou, Guanghong; Zhao, Gaiming; Li, Chunbao; Zhang, Yingjun; Chen, Lingzhen; Qi, Jun

    2009-03-01

    N-nitrosamines, biogenic amines and residual nitrite are harmful substances and often present in cured meat. The effects of gamma-irradiation (γ-irradiation) on these chemicals in dry-cured Chinese Rugao ham during ripening and post-ripening were investigated. Rugao hams were irradiated at a dose of 5kGy before ripening and were then ripened in an aging loft. Although γ-irradiation degraded tyramine, putrescine and spermine, on the other hand, it promoted the formation of spermidine, phenylethylamine, cadaverine and tryptamine. Residual nitrite was significantly reduced by γ-irradiation. N-nitrosodimethylamine (NDMA), N-nitrosodiethylamine (NDEA) and N-nitrosopyrrolidine (NPYR) were found in Chinese Rugao ham during ripening and post-ripening but could be degraded with γ-irradiation. The results suggest that γ-irradiation may be a potential decontamination measure for certain chemical compounds found in dry-cured meat.

  13. A comparative study of ripening among berries of the grape cluster reveals an altered transcriptional programme and enhanced ripening rate in delayed berries

    PubMed Central

    Gouthu, Satyanarayana; O’Neil, Shawn T.; Di, Yanming; Ansarolia, Mitra; Megraw, Molly; Deluc, Laurent G.

    2014-01-01

    Transcriptional studies in relation to fruit ripening generally aim to identify the transcriptional states associated with physiological ripening stages and the transcriptional changes between stages within the ripening programme. In non-climacteric fruits such as grape, all ripening-related genes involved in this programme have not been identified, mainly due to the lack of mutants for comparative transcriptomic studies. A feature in grape cluster ripening (Vitis vinifera cv. Pinot noir), where all berries do not initiate the ripening at the same time, was exploited to study their shifted ripening programmes in parallel. Berries that showed marked ripening state differences in a véraison-stage cluster (ripening onset) ultimately reached similar ripeness states toward maturity, indicating the flexibility of the ripening programme. The expression variance between these véraison-stage berry classes, where 11% of the genes were found to be differentially expressed, was reduced significantly toward maturity, resulting in the synchronization of their transcriptional states. Defined quantitative expression changes (transcriptional distances) not only existed between the véraison transitional stages, but also between the véraison to maturity stages, regardless of the berry class. It was observed that lagging berries complete their transcriptional programme in a shorter time through altered gene expressions and ripening-related hormone dynamics, and enhance the rate of physiological ripening progression. Finally, the reduction in expression variance of genes can identify new genes directly associated with ripening and also assess the relevance of gene activity to the phase of the ripening programme. PMID:25135520

  14. 5′-Methylthioadenosine Nucleosidase and 5-Methylthioribose Kinase Activities and Ethylene Production during Tomato Fruit Development and Ripening 1

    PubMed Central

    Kushad, Mosbah M.; Richardson, Daryl G.; Ferro, Adolph J.

    1985-01-01

    5′-Methylthioadenosine (MTA) nucleosidase and 5-methylthioribose (MTR) kinase activities were measured in crude extracts of tomato fruits (Lycopersicon esculentum Mill cv Rutgers) during fruit development and ripening. The highest activity of MTA nucleosidase (1.2 nanomoles per milligram protein per minute) was observed in small green fruits. The activity decreased during ripening; at the overripe stage only 6.5% of the peak activity remained. MTR kinase activity was low at the small green stage and increased thereafter until it reached peak activity at the breaker stage (0.7 nanomoles per milligram protein per minute) followed by a sharp decline at the later stages of fruit ripening. 1-Amino-cyclopropane-1-carboxylic acid (ACC) levels peaked at the red stage, while ethylene reached its highest level at the light-red stage. Several analogs of MTA and MTR were tested as both enzyme and ethylene inhibitors. Of the MTA analogs examined for their ability to inhibit MTA nucleosidase, 5′-chloroformycin reduced enzyme activity 89%, whereas 5′-chloroadenosine, 5′-isobutylthioadenosine, 5′-isopropylthioadenosine, and 5′-ethylthioadenosine inhibited the reaction with MTA by about 40%. 5′-Chloroformycin and 5′-chloroadenosine inhibited ethylene production over a period of 24 hours by about 64 and 42%, respectively. Other analogs of MTA were not effective inhibitors of ethylene production, whereas aminoethoxyvinylglycine showed a 34% inhibition over the same period of time. Of the MTR analogs tested, 5-isobutylthioribose was the most effective inhibitor of both MTR-kinase (41%) and ethylene production (35%). PMID:16664444

  15. Banana MaMADS Transcription Factors Are Necessary for Fruit Ripening and Molecular Tools to Promote Shelf-Life and Food Security1[OPEN

    PubMed Central

    Elitzur, Tomer; Yakir, Esther; Quansah, Lydia; Zhangjun, Fei; Vrebalov, Julia; Khayat, Eli; Giovannoni, James J.

    2016-01-01

    Genetic solutions to postharvest crop loss can reduce cost and energy inputs while increasing food security, especially for banana (Musa acuminata), which is a significant component of worldwide food commerce. We have functionally characterized two banana E class (SEPALLATA3 [SEP3]) MADS box genes, MaMADS1 and MaMADS2, homologous to the tomato (Solanum lycopersicum) RIN-MADS ripening gene. Transgenic banana plants repressing either gene (via antisense or RNA interference [RNAi]) were created and exhibited specific ripening delay and extended shelf-life phenotypes, including delayed color development and softening. The delay in fruit ripening is associated with a delay in climacteric respiration and reduced synthesis of the ripening hormone ethylene; in the most severe repressed lines, no ethylene was produced and ripening was most delayed. Unlike tomato rin mutants, banana fruits of all transgenic repression lines responded to exogenous ethylene by ripening normally, likely due to incomplete transgene repression and/or compensation by other MADS box genes. Our results show that, although MADS box ripening gene necessity is conserved across diverse taxa (monocots to dicots), unlike tomato, banana ripening requires at least two necessary members of the SEPALLATA MADS box gene group, and either can serve as a target for ripening control. The utility of such genes as tools for ripening control is especially relevant in important parthenocarpic crops such as the vegetatively propagated and widely consumed Cavendish banana, where breeding options for trait improvement are severely limited. PMID:26956665

  16. Malate synthase gene expression during fruit ripening of Cavendish banana (Musa acuminata cv. Williams).

    PubMed

    Pua, Eng-Chong; Chandramouli, Sumana; Han, Ping; Liu, Pei

    2003-01-01

    Malate synthase (MS) is a key enzyme responsible for malic acid synthesis in the glyoxylate cycle, which functions to convert stored lipids to carbohydrates, by catalysing the glyoxylate condensation reaction with acetyl-CoA in the peroxisome. In this study, the cloning of an MS cDNA, designated MaMS-1, from the banana fruit is reported. MaMS-1 was 1801 bp in length encoding a single polypeptide of 556 amino acid residues. Sequence analysis revealed that MaMS-1 possessed the conserved catalytic domain and a putative peroxisomal targeting signal SK(I/L) at the carboxyl terminal. MaMS-1 also shared an extensive sequence homology (79-81.3%) with other plant MS homologues. Southern analysis indicated that MS might be present as multiple members in the banana genome. In Northern analysis, MaMS-1 was expressed specifically in ripening fruit tissue and transcripts were not detected in other organs such as roots, pseudostem, leaves, ovary, male flower, and in fruit at different stages of development. However, the transcript abundance in fruit was affected by stage of ripening, during which transcript was barely detectable at the early stage of ripening (FG and TY), but the level increased markedly in MG and in other fruits at advanced ripening stages. Furthermore, MaMS-1 expression in FG fruit could be stimulated by treatment with 1 microl l(-1) exogenous ethylene, but the stimulatory effect was abolished by the application of an ethylene inhibitor, norbornadiene. Results of this study clearly show that MS expression in banana fruit is temporally regulated during ripening and is ethylene-inducible.

  17. Protein Synthesis in Relation to Ripening of Pome Fruits 1

    PubMed Central

    Frenkel, Chaim; Klein, Isaac; Dilley, D. R.

    1968-01-01

    Protein synthesis by intact Bartlett pear fruits was studied with ripening as measured by flesh softening, chlorophyll degradation, respiration, ethylene synthesis, and malic enzyme activity. Protein synthesis is required for normal ripening, and the proteins synthesized early in the ripening process are, in fact, enzymes required for ripening. 14C-Phenylalanine is differentially incorporated into fruit proteins separated by acrylamide gel electrophoresis of pome fruits taken at successive ripening stages. Capacity for malic enzyme synthesis increases during the early stage of ripening. Fruit ripening and ethylene synthesis are inhibited when protein synthesis is blocked by treatment with cycloheximide at the early-climacteric stage. Cycloheximide became less effective as the climacteric developed. Ethylene did not overcome inhibition of ripening by cycloheximide. The respiratory climacteric is not inhibited by cycloheximide. It is concluded that normal ripening of pome fruits is a highly coordinated process of biochemical differentiation involving directed protein synthesis. PMID:16656897

  18. Berry Flesh and Skin Ripening Features in Vitis vinifera as Assessed by Transcriptional Profiling

    PubMed Central

    Grimplet, Jérôme; Bravo, Gema; Flores, Pilar; Fenoll, José; Hellín, Pilar; Oliveros, Juan Carlos; Martínez-Zapater, José M.

    2012-01-01

    Background Ripening of fleshy fruit is a complex developmental process involving the differentiation of tissues with separate functions. During grapevine berry ripening important processes contributing to table and wine grape quality take place, some of them flesh- or skin-specific. In this study, transcriptional profiles throughout flesh and skin ripening were followed during two different seasons in a table grape cultivar ‘Muscat Hamburg’ to determine tissue-specific as well as common developmental programs. Methodology/Principal Findings Using an updated GrapeGen Affymetrix GeneChip® annotation based on grapevine 12×v1 gene predictions, 2188 differentially accumulated transcripts between flesh and skin and 2839 transcripts differentially accumulated throughout ripening in the same manner in both tissues were identified. Transcriptional profiles were dominated by changes at the beginning of veraison which affect both pericarp tissues, although frequently delayed or with lower intensity in the skin than in the flesh. Functional enrichment analysis identified the decay on biosynthetic processes, photosynthesis and transport as a major part of the program delayed in the skin. In addition, a higher number of functional categories, including several related to macromolecule transport and phenylpropanoid and lipid biosynthesis, were over-represented in transcripts accumulated to higher levels in the skin. Functional enrichment also indicated auxin, gibberellins and bHLH transcription factors to take part in the regulation of pre-veraison processes in the pericarp, whereas WRKY and C2H2 family transcription factors seems to more specifically participate in the regulation of skin and flesh ripening, respectively. Conclusions/Significance A transcriptomic analysis indicates that a large part of the ripening program is shared by both pericarp tissues despite some components are delayed in the skin. In addition, important tissue differences are present from early

  19. Germination response in wheat grains to dihydroactinidiolide, a germination inhibitor in wheat husks, and related compounds.

    PubMed

    Kato, Tadahiro; Imai, Takahito; Kashimura, Kaori; Saito, Naoko; Masaya, Kengo

    2003-04-09

    On the basis of our recent findings that the germination of intact wheat grains with glumes (husks) belonging to dormant varieties was restrained as compared with that of dehusked grains, we have explored the identities of germination inhibitors in the glumes, resulting in the characterization of dihydroactinidiolide (1) and some aromatic compounds. A related natural product, tetrahydroactinidiolide (2), showed similar activity. The present study has demonstrated that the sensitivity in inhibition response of germination of the grains to 1 and 2 declined during after-ripening, in parallel with changes in germinability; the sprouting of after-ripened seeds on a whole spike was preventable by exogenous application of 2 in laboratory conditions, and germination of after-ripened grains was delayed by more than two weeks by the action of 1 or 2. The term "pseudodormancy" is proposed for the phenomenon of delay of germination caused by the inhibitor. After accumulation of additional evidence on inhibition response of actinidiolide-type natural products, structurally related to inhibitor 1, a mechanism concerning germination inhibition by 1 or 2 is proposed on the basis of the concept of nonbonding interaction with the inhibitors at an active site of an acceptor.

  20. Comprehensive ripeness-index for prediction of ripening level in mangoes by multivariate modelling of ripening behaviour

    NASA Astrophysics Data System (ADS)

    Eyarkai Nambi, Vijayaram; Thangavel, Kuladaisamy; Manickavasagan, Annamalai; Shahir, Sultan

    2017-01-01

    Prediction of ripeness level in climacteric fruits is essential for post-harvest handling. An index capable of predicting ripening level with minimum inputs would be highly beneficial to the handlers, processors and researchers in fruit industry. A study was conducted with Indian mango cultivars to develop a ripeness index and associated model. Changes in physicochemical, colour and textural properties were measured throughout the ripening period and the period was classified into five stages (unripe, early ripe, partially ripe, ripe and over ripe). Multivariate regression techniques like partial least square regression, principal component regression and multi linear regression were compared and evaluated for its prediction. Multi linear regression model with 12 parameters was found more suitable in ripening prediction. Scientific variable reduction method was adopted to simplify the developed model. Better prediction was achieved with either 2 or 3 variables (total soluble solids, colour and acidity). Cross validation was done to increase the robustness and it was found that proposed ripening index was more effective in prediction of ripening stages. Three-variable model would be suitable for commercial applications where reasonable accuracies are sufficient. However, 12-variable model can be used to obtain more precise results in research and development applications.

  1. Perturbation of polyamine catabolism affects grape ripening of Vitis vinifera cv. Trincadeira.

    PubMed

    Agudelo-Romero, Patricia; Ali, Kashif; Choi, Young H; Sousa, Lisete; Verpoorte, Rob; Tiburcio, Antonio F; Fortes, Ana M

    2014-01-01

    Grapes are economically the most important fruit worldwide. However, the complexity of biological events that lead to ripening of nonclimacteric fruits is not fully understood, particularly the role of polyamines' catabolism. The transcriptional and metabolic profilings complemented with biochemical data were studied during ripening of Trincadeira grapes submitted to guazatine treatment, a potent inhibitor of polyamine oxidase activity. The mRNA expression profiles of one time point (EL 38) corresponding to harvest stage was compared between mock and guazatine treatments using Affymetrix GrapeGen(®) genome array. A total of 2113 probesets (1880 unigenes) were differentially expressed between these samples. Quantitative RT-PCR validated microarrays results being carried out for EL 35 (véraison berries), EL 36 (ripe berries) and EL 38 (harvest stage berries). Metabolic profiling using HPLC and (1)H NMR spectroscopy showed increase of putrescine, proline, threonine and 1-O-ethyl-β-glucoside in guazatine treated samples. Genes involved in amino acid, carbohydrate and water transport were down-regulated in guazatine treated samples suggesting that the strong dehydrated phenotype obtained in guazatine treated samples may be due to impaired transport mechanisms. Genes involved in terpenes' metabolism were differentially expressed between guazatine and mock treated samples. Altogether, results support an important role of polyamine catabolism in grape ripening namely in cell expansion and aroma development. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  2. Residual effects of low oxygen storage of mature green fruit on ripening processes and ester biosynthesis during ripening in bananas

    USDA-ARS?s Scientific Manuscript database

    Mature green banana (Musa sapientum L. cv. Cavendish) fruit were stored in 0.5%, 2 %, or 21% O2 for 7 days at 20 °C before ripening was initiated by ethylene. Residual effects of low O2 storage in mature green fruit on ripening and ester biosynthesis in fruit were investigated during ripening period...

  3. Developmental gene regulation during tomato fruit ripening and in-vitro sepal morphogenesis

    PubMed Central

    Bartley, Glenn E; Ishida, Betty K

    2003-01-01

    Background Red ripe tomatoes are the result of numerous physiological changes controlled by hormonal and developmental signals, causing maturation or differentiation of various fruit tissues simultaneously. These physiological changes affect visual, textural, flavor, and aroma characteristics, making the fruit more appealing to potential consumers for seed dispersal. Developmental regulation of tomato fruit ripening has, until recently, been lacking in rigorous investigation. We previously indicated the presence of up-regulated transcription factors in ripening tomato fruit by data mining in TIGR Tomato Gene Index. In our in-vitro system, green tomato sepals cultured at 16 to 22°C turn red and swell like ripening tomato fruit while those at 28°C remain green. Results Here, we have further examined regulation of putative developmental genes possibly involved in tomato fruit ripening and development. Using molecular biological methods, we have determined the relative abundance of various transcripts of genes during in vitro sepal ripening and in tomato fruit pericarp at three stages of development. A number of transcripts show similar expression in fruits to RIN and PSY1, ripening-associated genes, and others show quite different expression. Conclusions Our investigation has resulted in confirmation of some of our previous database mining results and has revealed differences in gene expression that may be important for tomato cultivar variation. We present new and intriguing information on genes that should now be studied in a more focused fashion. PMID:12906715

  4. Thermotolerance responses in ripening berries of Vitis vinifera L. cv Muscat Hamburg.

    PubMed

    Carbonell-Bejerano, Pablo; Santa María, Eva; Torres-Pérez, Rafael; Royo, Carolina; Lijavetzky, Diego; Bravo, Gema; Aguirreolea, Jone; Sánchez-Díaz, Manuel; Antolín, M Carmen; Martínez-Zapater, José M

    2013-07-01

    Berry organoleptic properties are highly influenced by ripening environmental conditions. In this study, we used grapevine fruiting cuttings to follow berry ripening under different controlled conditions of temperature and irradiation intensity. Berries ripened at higher temperatures showed reduced anthocyanin accumulation and hastened ripening, leading to a characteristic drop in malic acid and total acidity. The GrapeGen GeneChip® combined with a newly developed GrapeGen 12Xv1 MapMan version were utilized for the functional analysis of berry transcriptomic differences after 2 week treatments from veraison onset. These analyses revealed the establishment of a thermotolerance response in berries under high temperatures marked by the induction of heat shock protein (HSP) chaperones and the repression of transmembrane transporter-encoding transcripts. The thermotolerance response was coincident with up-regulation of ERF subfamily transcription factors and increased ABA levels, suggesting their participation in the maintenance of the acclimation response. Lower expression of amino acid transporter-encoding transcripts at high temperature correlated with balanced amino acid content, suggesting a transcriptional compensation of temperature effects on protein and membrane stability to allow for completion of berry ripening. In contrast, the lower accumulation of anthocyanins and higher malate metabolization measured under high temperature might partly result from imbalance in the expression and function of their specific transmembrane transporters and expression changes in genes involved in their metabolic pathways. These results open up new views to improve our understanding of berry ripening under high temperatures.

  5. Critical roles of DNA demethylation in the activation of ripening-induced genes and inhibition of ripening-repressed genes in tomato fruit

    PubMed Central

    Lang, Zhaobo; Wang, Yihai; Tang, Kai; Tang, Dengguo; Datsenka, Tatsiana; Cheng, Jingfei; Zhang, Yijing; Handa, Avtar K.

    2017-01-01

    DNA methylation is a conserved epigenetic mark important for genome integrity, development, and environmental responses in plants and mammals. Active DNA demethylation in plants is initiated by a family of 5-mC DNA glycosylases/lyases (i.e., DNA demethylases). Recent reports suggested a role of active DNA demethylation in fruit ripening in tomato. In this study, we generated loss-of-function mutant alleles of a tomato gene, SlDML2, which is a close homolog of the Arabidopsis DNA demethylase gene ROS1. In the fruits of the tomato mutants, increased DNA methylation was found in thousands of genes. These genes included not only hundreds of ripening-induced genes but also many ripening-repressed genes. Our results show that SlDML2 is critical for tomato fruit ripening and suggest that active DNA demethylation is required for both the activation of ripening-induced genes and the inhibition of ripening-repressed genes. PMID:28507144

  6. Genome-wide analysis of the NAC transcription factor family and their expression during the development and ripening of the Fragaria × ananassa fruits

    PubMed Central

    Matas-Arroyo, Antonio J.; Caballero, José Luis; Muñoz-Blanco, Juan

    2018-01-01

    NAC proteins are a family of transcription factors which have a variety of important regulatory roles in plants. They present a very well conserved group of NAC subdomains in the N-terminal region and a highly variable domain at the C-terminus. Currently, knowledge concerning NAC family in the strawberry plant remains very limited. In this work, we analyzed the NAC family of Fragaria vesca, and a total of 112 NAC proteins were identified after we curated the annotations from the version 4.0.a1 genome. They were placed into the ligation groups (pseudo-chromosomes) and described its physicochemical and genetic features. A microarray transcriptomic analysis showed six of them expressed during the development and ripening of the Fragaria x ananassa fruit. Their expression patterns were studied in fruit (receptacle and achenes) in different stages of development and in vegetative tissues. Also, the expression level under different hormonal treatments (auxins, ABA) and drought stress was investigated. In addition, they were clustered with other NAC transcription factor with known function related to growth and development, senescence, fruit ripening, stress response, and secondary cell wall and vascular development. Our results indicate that these six strawberry NAC proteins could play different important regulatory roles in the process of development and ripening of the fruit, providing the basis for further functional studies and the selection for NAC candidates suitable for biotechnological applications. PMID:29723301

  7. Analysis of ripening-related gene expression in papaya using an Arabidopsis-based microarray

    PubMed Central

    2012-01-01

    Background Papaya (Carica papaya L.) is a commercially important crop that produces climacteric fruits with a soft and sweet pulp that contain a wide range of health promoting phytochemicals. Despite its importance, little is known about transcriptional modifications during papaya fruit ripening and their control. In this study we report the analysis of ripe papaya transcriptome by using a cross-species (XSpecies) microarray technique based on the phylogenetic proximity between papaya and Arabidopsis thaliana. Results Papaya transcriptome analyses resulted in the identification of 414 ripening-related genes with some having their expression validated by qPCR. The transcription profile was compared with that from ripening tomato and grape. There were many similarities between papaya and tomato especially with respect to the expression of genes encoding proteins involved in primary metabolism, regulation of transcription, biotic and abiotic stress and cell wall metabolism. XSpecies microarray data indicated that transcription factors (TFs) of the MADS-box, NAC and AP2/ERF gene families were involved in the control of papaya ripening and revealed that cell wall-related gene expression in papaya had similarities to the expression profiles seen in Arabidopsis during hypocotyl development. Conclusion The cross-species array experiment identified a ripening-related set of genes in papaya allowing the comparison of transcription control between papaya and other fruit bearing taxa during the ripening process. PMID:23256600

  8. Cloning of an ADP-ribosylation factor gene from banana (Musa acuminata) and its expression patterns in postharvest ripening fruit.

    PubMed

    Wang, Yuan; Wu, Jing; Xu, Bi-Yu; Liu, Ju-Hua; Zhang, Jian-Bin; Jia, Cai-Hong; Jin, Zhi-Qiang

    2010-08-15

    A full-length cDNA encoding an ADP-ribosylation factor (ARF) from banana (Musa acuminata) fruit was cloned and named MaArf. It contains an open reading frame encoding a 181-amino-acid polypeptide. Sequence analysis showed that MaArf shared high similarity with ARF of other plant species. The genomic sequence of MaArf was also obtained using polymerase chain reaction (PCR). Sequence analysis showed that MaArf was a split gene containing five exons and four introns in genomic DNA. Reverse-transcriptase PCR was used to analyze the spatial expression of MaArf. The results showed that MaArf was expressed in all the organs examined: root, rhizome, leaf, flower and fruit. Real-time quantitative PCR was used to explore expression patterns of MaArf in postharvest banana. There was differential expression of MaArf associated with ethylene biosynthesis. In naturally ripened banana, expression of MaArf was in accordance with ethylene biosynthesis. However, in 1-methylcyclopropene-treated banana, the expression of MaArf was inhibited and changed little. When treated with ethylene, MaArf expression in banana fruit significantly increased in accordance with ethylene biosynthesis; the peak of MaArf was 3 d after harvest, 11 d earlier than for naturally ripened banana fruits. These results suggest that MaArf is induced by ethylene in regulating postharvest banana ripening. Finally, subcellular localization assays showed the MaArf protein in the cytoplasm. Copyright 2010 Elsevier GmbH. All rights reserved.

  9. Apple (Malus domestica) MdERF2 negatively affects ethylene biosynthesis during fruit ripening by suppressing MdACS1 transcription.

    PubMed

    Li, Tong; Jiang, Zhongyu; Zhang, Lichao; Tan, Dongmei; Wei, Yun; Yuan, Hui; Li, Tianlai; Wang, Aide

    2016-12-01

    Ripening in climacteric fruit requires the gaseous phytohormone ethylene. Although ethylene signaling has been well studied, knowledge of the transcriptional regulation of ethylene biosynthesis is still limited. Here we show that an apple (Malus domestica) ethylene response factor, MdERF2, negatively affects ethylene biosynthesis and fruit ripening by suppressing the transcription of MdACS1, a gene that is critical for biosynthesis of ripening-related ethylene. Expression of MdERF2 was suppressed by ethylene during ripening of apple fruit, and we observed that MdERF2 bound to the promoter of MdACS1 and directly suppressed its transcription. Moreover, MdERF2 suppressed the activity of the promoter of MdERF3, a transcription factor that we found to bind to the MdACS1 promoter, thereby increasing MdACS1 transcription. We determined that the MdERF2 and MdERF3 proteins directly interact, and this interaction suppresses the binding of MdERF3 to the MdACS1 promoter. Moreover, apple fruit with transiently downregulated MdERF2 expression showed higher ethylene production and faster ripening. Our results indicate that MdERF2 negatively affects ethylene biosynthesis and fruit ripening in apple by suppressing the transcription of MdACS1 via multiple mechanisms, thereby acting as an antagonist of positive ripening regulators. Our findings offer a deep understanding of the transcriptional regulation of ethylene biosynthesis during climacteric fruit ripening. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

  10. Influence of cold storage prior to and after ripening on quality factors and sensory attributes of ‘Hass’ avocados

    USDA-ARS?s Scientific Manuscript database

    Partially-ripened avocados are often held in cold storage in an attempt to enable the consistent delivery of ripe fruit to food service or retail outlets, although the effect on the quality of such fruit is incompletely understood. ‘Hass’ avocados were ripened to near ripeness (13.3 - 17.8 N) at 20 ...

  11. Risk factors for inhibitor development in severe hemophilia a.

    PubMed

    Garagiola, Isabella; Palla, Roberta; Peyvandi, Flora

    2018-05-25

    Although significant advances in hemophilia treatment have improved patient outcomes and quality of life, one of the greatest complications in severe hemophilia A is the development of anti-Factor VIII (FVIII) antibodies that inhibit FVIII activity in almost 30% of previously untreated patients (PUPs). Inhibitors make very difficult the management of patients and increase their morbidity and mortality reducing drastically their quality of life. Numerous studies have investigated the mechanisms leading to the development of FVIII inhibitors. However, the etiology of their onset is complex and not yet fully understood. Inhibitors develop from a multicausal immune response involving both genetic (unmodifiable) and environmental (modifiable) factors. F8 gene mutations are the most important genetic risk factor, with null mutations being associated with the highest risk of inhibitor development. Immune response genes (e.g. the human leukocyte antigen complex) and proteins (e.g. cytokines) were studied without any strong confirmation of their role in modulating of inhibitor development. Type of FVIII product is the most important modifiable risk factor. The plasma-derived products containing von Willebrand factor were recently suggested to determine a lower incidence of inhibitor development than recombinant products in PUPs, in the first 50 exposure days (EDs). Other environmental factors including, age at first treatment, treatment intensity and the danger signal effect (surgery, severe bleeds, vaccinations and infections) has also been postulated as an explanation for environment-related inhibitor risk. This review reports the current knowledge on genetic and environmental risk factors on inhibitor development in patients with severe hemophilia A. Copyright © 2018 Elsevier Ltd. All rights reserved.

  12. Artificial ripening of sugar pine seeds

    Treesearch

    Stanley L. Krugman

    1966-01-01

    Immature sugar pine seeds were collected and ripened either in the cone or in moist vermiculate. Seeds collected before the second week of August could not be artificially ripened and the causes for these failures were investigated. After the second week of August, immature seeds could be brought to maturity. A practical method for a commercial operation should be...

  13. Textural properties of mango cultivars during ripening.

    PubMed

    Jha, Shyam Narayan; Jaiswal, Pranita; Narsaiah, Kairam; Kaur, Poonam Preet; Singh, Ashish Kumar; Kumar, Ramesh

    2013-12-01

    Firmness and toughness of fruit, peel and pulp of seven different mango cultivars were studied over a ripening period of ten days to investigate the effects of harvesting stages (early, mid and late) on fruit quality. Parameters were measured at equatorial region of fruits using TA-Hdi Texture Analyzer. The textural characteristics showed a rapid decline in their behaviour until mangoes got ripened and thereafter, the decline became almost constant indicating the completion of ripening. However, the rate of decline in textural properties was found to be cultivar specific. In general, the changes in textural attributes were found to be significantly influenced by ripening period and stage of harvesting, but firmness attributes (peel, fruit and pulp) of early harvested mangoes did not differ significantly from mid harvested mangoes, while peel, fruit and pulp firmness of late harvested mangoes were found to be significantly lower than early and mid harvested mangoes.

  14. Camembert-type cheese ripening dynamics are changed by the properties of wrapping films.

    PubMed

    Picque, D; Leclercq-Perlat, M N; Guillemin, H; Perret, B; Cattenoz, T; Provost, J J; Corrieu, G

    2010-12-01

    Four gas-permeable wrapping films exhibiting different degrees of water permeability (ranging from 1.6 to 500 g/m(2) per d) were tested to study their effect on soft-mold (Camembert-type) cheese-ripening dynamics compared with unwrapped cheeses. Twenty-three-day trials were performed in 2 laboratory-size (18L) respiratory-ripening cells under controlled temperature (6 ± 0.5°C), relative humidity (75 ± 2%), and carbon dioxide content (0.5 to 1%). The films allowed for a high degree of respiratory activity; no limitation in gas permeability was observed. The wide range of water permeability of the films led to considerable differences in cheese water loss (from 0.5 to 12% on d 23, compared with 15% for unwrapped cheeses), which appeared to be a key factor in controlling cheese-ripening progress. A new relationship between 2 important cheese-ripening descriptors (increase of the cheese core pH and increase of the cheese's creamy underrind thickness) was shown in relation to the water permeability of the wrapping film. High water losses (more than 10 to 12% on d 23) also were observed for unwrapped cheeses, leading to Camembert cheeses that were too dry and poorly ripened. On the other hand, low water losses (from 0.5 to 1% on d 23) led to over-ripening in the cheese underrind, which became runny as a result. Finally, water losses from around 3 to 6% on d 23 led to good ripening dynamics and the best cheese quality. This level of water loss appeared to be ideal in terms of cheese-wrapping film design. Copyright © 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  15. Differential gene expression in ripening banana fruit.

    PubMed Central

    Clendennen, S K; May, G D

    1997-01-01

    During banana (Musa acuminata L.) fruit ripening ethylene production triggers a developmental cascade that is accompanied by a massive conversion of starch to sugars, an associated burst of respiratory activity, and an increase in protein synthesis. Differential screening of cDNA libraries representing banana pulp at ripening stages 1 and 3 has led to the isolation of 11 nonredundant groups of differentially expressed mRNAs. Identification of these transcripts by partial sequence analysis indicates that two of the mRNAs encode proteins involved in carbohydrate metabolism, whereas others encode proteins thought to be associated with pathogenesis, senescence, or stress responses in plants. Their relative abundance in the pulp and tissue-specific distribution in greenhouse-grown banana plants were determined by northern-blot analyses. The relative abundance of transcripts encoding starch synthase, granule-bound starch synthase, chitinase, lectin, and a type-2 metallothionein decreased in pulp during ripening. Transcripts encoding endochitinase, beta-1,3-glucanase, a thaumatin-like protein, ascorbate peroxidase, metallothionein, and a putative senescence-related protein increased early in ripening. The elucidation of the molecular events associated with banana ripening will facilitate a better understanding and control of these processes, and will allow us to attain our long-term goal of producing candidate oral vaccines in transgenic banana plants. PMID:9342866

  16. The ambiguous ripening nature of the fig (Ficus carica L.) fruit: a gene-expression study of potential ripening regulators and ethylene-related genes

    PubMed Central

    Freiman, Zohar E.; Rosianskey, Yogev; Dasmohapatra, Rajeswari; Kamara, Itzhak; Flaishman, Moshe A.

    2015-01-01

    The traditional definition of climacteric and non-climacteric fruits has been put into question. A significant example of this paradox is the climacteric fig fruit. Surprisingly, ripening-related ethylene production increases following pre- or postharvest 1-methylcyclopropene (1-MCP) application in an unexpected auto-inhibitory manner. In this study, ethylene production and the expression of potential ripening-regulator, ethylene-synthesis, and signal-transduction genes are characterized in figs ripening on the tree and following preharvest 1-MCP application. Fig ripening-related gene expression was similar to that in tomato and apple during ripening on the tree, but only in the fig inflorescence–drupelet section. Because the pattern in the receptacle is different for most of the genes, the fig drupelets developed inside the syconium are proposed to function as parthenocarpic true fruit, regulating ripening processes for the whole accessory fruit. Transcription of a potential ripening regulator, FcMADS8, increased during ripening on the tree and was inhibited following 1-MCP treatment. Expression patterns of the ethylene-synthesis genes FcACS2, FcACS4, and FcACO3 could be related to the auto-inhibition reaction of ethylene production in 1-MCP-treated fruit. Along with FcMADS8 suppression, gene expression analysis revealed upregulation of FcEBF1, and downregulation of FcEIL3 and several FcERFs by 1-MCP treatment. This corresponded with the high storability of the treated fruit. One FcERF was overexpressed in the 1-MCP-treated fruit, and did not share the increasing pattern of most FcERFs in the tree-ripened fig. This demonstrates the potential of this downstream ethylene-signal-transduction component as an ethylene-synthesis regulator, responsible for the non-climacteric auto-inhibition of ethylene production in fig. PMID:25956879

  17. Factors affecting ethylene and carbon dioxide concentrations during ripening: Incidence on final dry matter, total soluble solids content and acidity of mango fruit.

    PubMed

    Nordey, Thibault; Léchaudel, Mathieu; Génard, Michel; Joas, Jacques

    2016-06-01

    Ripening of climacteric fruits is associated with pronounced changes in fruit gas composition caused by a concomitant rise in respiration and ethylene production. There is a discrepancy in the literature since some authors reported that changes in fruit gas compositions differ in attached and detached fruits. This study presents for the first time an overview of pre- and post-harvest factors that lead to variations in the climacteric respiration and ethylene production, and attempts to determine their impacts on fruit composition, i.e., dry matter, total soluble solids content and acidity. The impact of growing conditions such as the fruit position in the canopy and the fruit carbon supply; fruit detachment from the tree, including the maturity stage at harvest; and storage conditions after harvest, i.e., relative humidity and temperature were considered as well as changes in fruit skin resistance to gas diffusion during fruit growth and storage. Results showed that fruit gas composition vary with all pre and post-harvest factors studied. Although all mangoes underwent a respiratory climacteric and an autocatalytic ethylene production, whatever pre and post-harvest factors studied, large differences in ethylene production, climacteric respiration and fruit quality were measured. Results suggested that the ripening capacity is not related to the fruit ability to produce great amount of ethylene. In agreement with precedent studies, this work provided several lines of evidence that gas composition of fruit is related to its water balance. Our measurements indicated that skin resistance to gas diffusion increased after the harvest and during storage. It was so suggested that the faster ripening of detached fruit may be explained in part by changes in fruit water balance and skin resistance to gas diffusion caused by fruit detachment. Copyright © 2016 Elsevier GmbH. All rights reserved.

  18. Lipolytic and oxidative changes in 'Chorizo' during ripening.

    PubMed

    Fernández, M C; Rodríguez, J M

    1991-01-01

    Changes in fats during the ripening of 'chorizo'-a Spanish dry sausage- elaborated by traditional and industrial processes have been studied. Total free fatty acids (FFA), carbonyls, volatile fatty acids (acetic acid), TBA number and individual FFA were determined. A marked increase of total free fatty acids was observed, although an increase of rancidity was not detected. Except in one of the batches studied in which a rise in carbonyls was reported, these compounds remained unchanged during the ripening in the rest of the batches analysed. No significant change occurred in the proportions of different free fatty acids during ripening in both types of 'chorizo'. There was a marked increase of acetic acid during the first days of ripening followed by a slow fall in the ultimate stages. Copyright © 1990. Published by Elsevier Ltd.

  19. 21 CFR 133.182 - Soft ripened cheeses.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Soft ripened cheeses. 133.182 Section 133.182 Food... HUMAN CONSUMPTION CHEESES AND RELATED CHEESE PRODUCTS Requirements for Specific Standardized Cheese and Related Products § 133.182 Soft ripened cheeses. (a) The cheeses for which definitions and standards of...

  20. Changes in polysaccharide and protein composition of cell walls in grape berry skin (Cv. Shiraz) during ripening and over-ripening.

    PubMed

    Vicens, Anysia; Fournand, David; Williams, Pascale; Sidhoum, Louise; Moutounet, Michel; Doco, Thierry

    2009-04-08

    Polysaccharide modification is the most fundamental factor that affects firmness of fruit during ripening. In grape, because of the lack of information on the modifications occurring in cell wall polysaccharides in skins, but also because this tissue contains large amounts of organoleptic compounds for winemaking, a study was performed on the evolution and extractability of polysaccharides from grape skins of Shiraz cultivar throughout ripening. A HEPES/phenol extraction technique was used to analyze Shiraz grape cell wall material isolated from skins of berries harvested from one to ten weeks after veraison. Total amounts in cell wall polysaccharides remained constant during ripening (4.2 mg/berry). A slight decrease in galactose content of insoluble polysaccharides was observed, as well as a significant de-esterification of methoxylated uronic acids, indicating that some modifications occur in cell wall polysaccharides. The water-soluble fraction represented a very small fraction of the whole polysaccharides, but its amounts increased more than 2-fold between the first and the last sample. Isolated cell walls were also analyzed for their protein composition. Last, hydroalcoholic extractions in model-wine solution were also performed on fresh skins. This extracted fraction was very similar to the water-soluble one, and increased during the entire period. By comparison with polysaccharide modifications described in flesh cell wall in previous works, it can be assumed that the moderate skin polysaccharide degradation highlights the protective role of that tissue.

  1. Fibroblast growth factor receptor inhibitors.

    PubMed

    Kumar, Suneel B V S; Narasu, Lakshmi; Gundla, Rambabu; Dayam, Raveendra; J A R P, Sarma

    2013-01-01

    Fibroblast growth factor receptors (FGFRs) play an important role in embryonic development, angiogenesis, wound healing, cell proliferation and differentiation. The fibroblast growth factor receptor (FGFR) isoforms have been under intense scrutiny for effective anticancer drug candidates. The fibroblast growth factor (FGF) and its receptor (FGFR) provide another pathway that seems critical to monitoring angiogenesis. Recent findings suggest that FGFR mediates signaling, regulates the PKM2 activity, and plays a crucial role in cancer metabolism. The current review also covers the recent findings on the role of FGFR1 in cancer metabolism. This paper reviews the progress, mechanism, and binding modes of recently known kinase inhibitors such as PD173074, SU series and other inhibitors still under clinical development. Some of the structural classes that will be highlighted in this review include Pyrido[2,3-d]pyrimidines, Indolin- 2-one, Pyrrolo[2,1-f][1,2,4]triazine, Pyrido[2,3-d]pyrimidin-7(8H)-one, and 1,6- Naphthyridin-2(1H)-ones.

  2. Suppression of Ostwald Ripening by Chemical Reactions

    NASA Astrophysics Data System (ADS)

    Zwicker, David; Hyman, Anthony A.; Jülicher, Frank

    2015-03-01

    Emulsions consisting of droplets immersed in a fluid are typically unstable and coarsen over time. One important coarsening process is Ostwald ripening, which is driven by the surface tension of the droplets. Ostwald ripening must thus be suppressed to stabilize emulsions, e.g. to control the properties of pharmaceuticals, food, or cosmetics. Suppression of Ostwald ripening is also important in biological cells, which contain stable liquid-like compartments, e.g. germ granules, Cajal-bodies, and centrosomes. Such systems are often driven away from equilibrium by chemical reactions and can thus be called active emulsions. Here, we show that non-equilibrium chemical reactions can suppress Ostwald Ripening, leading to stable, monodisperse emulsions. We derive analytical approximations of the typical droplet size, droplet count, and time scale of the dynamics from a coarse-grained description of the droplet dynamics. We also compare these results to numerical simulations of the continuous concentration fields. Generally, we thus show how chemical reactions can be used to stabilize emulsions and to control their properties in technology and nature.

  3. Crystallization mechanisms in cream during ripening and initial butter churning.

    PubMed

    Buldo, Patrizia; Kirkensgaard, Jacob J K; Wiking, Lars

    2013-01-01

    The temperature treatment of cream is the time-consuming step in butter production. A better understanding of the mechanisms leading to partial coalescence, such as fat crystallization during ripening and churning of the cream, will contribute to optimization of the production process. In this study, ripening and churning of cream were performed in a rheometer cell and the mechanisms of cream crystallization during churning of the cream, including the effect of ripening time, were investigated to understand how churning time and partial coalescence are affected. Crystallization mechanisms were studied as function of time by differential scanning calorimetry, nuclear magnetic resonance and by X-ray scattering. Microstructure formation was investigated by small deformation rheology and static light scattering. The study demonstrated that viscosity measurements can be used to detect phase inversion of the emulsion during churning of the cream in a rheometer cell. Longer ripening time (e.g., 5h vs. 0 h) resulted in larger butter grains (91 vs. 52 µm), higher viscosity (5.3 vs. 1.3 Pa · s), and solid fat content (41 vs. 13%). Both ripening and churning time had an effect on the thermal behavior of the cream. Despite the increase in solid fat content, no further changes in crystal polymorphism and in melting behavior were observed after 1h of ripening and after churning. The churning time significantly decreased after 0.5h of ripening, from 22.9 min for the cream where no ripening was applied to 16.23 min. Therefore, the crystallization state that promotes partial coalescence (i.e., aggregation of butter grains) is obtained within the first hour of cream ripening at 10 °C. The present study adds knowledge on the fundamental processes of crystallization and polymorphism of milk fat occurring during ripening and churning of cream. In addition, the dairy industry will benefit from these insights on the optimization of butter manufacturing. Copyright © 2013 American

  4. Surface Microflora of Four Smear-Ripened Cheeses

    PubMed Central

    Mounier, Jérôme; Gelsomino, Roberto; Goerges, Stefanie; Vancanneyt, Marc; Vandemeulebroecke, Katrien; Hoste, Bart; Scherer, Siegfried; Swings, Jean; Fitzgerald, Gerald F.; Cogan, Timothy M.

    2005-01-01

    The microbial composition of smear-ripened cheeses is not very clear. A total of 194 bacterial isolates and 187 yeast isolates from the surfaces of four Irish farmhouse smear-ripened cheeses were identified at the midpoint of ripening using pulsed-field gel electrophoresis (PFGE), repetitive sequence-based PCR, and 16S rRNA gene sequencing for identifying and typing the bacteria and Fourier transform infrared spectroscopy and mitochondrial DNA restriction fragment length polymorphism (mtDNA RFLP) analysis for identifying and typing the yeast. The yeast microflora was very uniform, and Debaryomyces hansenii was the dominant species in the four cheeses. Yarrowia lipolytica was also isolated in low numbers from one cheese. The bacteria were highly diverse, and 14 different species, Corynebacterium casei, Corynebacterium variabile, Arthrobacter arilaitensis, Arthrobacter sp., Microbacterium gubbeenense, Agrococcus sp. nov., Brevibacterium linens, Staphylococcus epidermidis, Staphylococcus equorum, Staphylococcus saprophyticus, Micrococcus luteus, Halomonas venusta, Vibrio sp., and Bacillus sp., were identified on the four cheeses. Each cheese had a more or less unique microflora with four to nine species on its surface. However, two bacteria, C. casei and A. arilaitensis, were found on each cheese. Diversity at the strain level was also observed, based on the different PFGE patterns and mtDNA RFLP profiles of the dominant bacterial and yeast species. None of the ripening cultures deliberately inoculated onto the surface were reisolated from the cheeses. This study confirms the importance of the adventitious, resident microflora in the ripening of smear cheeses. PMID:16269673

  5. Brucella melitensis survival during manufacture of ripened goat cheese at two temperatures.

    PubMed

    Méndez-González, Karla Y; Hernández-Castro, Rigoberto; Carrillo-Casas, Erika M; Monroy, Jorge F; López-Merino, Ahide; Suárez-Güemes, Francisco

    2011-12-01

    The aim of the current work was to assess the influence of two temperatures, 4°C and 24°C, on pH and water activity and their association with Brucella melitensis survival during the traditional manufacture of ripened goat cheese. Raw milk from a brucellosis-free goat herd was used for the manufacture of ripened cheese. The cheese was inoculated with 5×10(9) of the B. melitensis 16M strain during the tempering stage. The cheeses were matured for 5, 20, and 50 days at both temperatures. To assess Brucella survival, the pH and a(w) were recorded at each stage of the process (curd cutting, draining whey, immersion in brine, ripening I, ripening II, and ripening III). B. melitensis was detected at ripening stage III (1×10(3) colony-forming unit [CFU]/mL) from cheeses matured at 4°C with a pH of 5.0 and a(w) of 0.90, and at a ripening stage II (1×10(4) CFU/mL) from cheeses ripened at 24°C with a pH of 4.0 and a(w) of 0.89. The remaining stages were free from the inoculated pathogen. In addition, viable B. melitensis was recovered in significant amounts (1-2×10(6) CFU/mL) from the whey fractions of both types of cheese ripened at 24°C and 4°C. These results revealed the effects of high temperature (24°C vs. 4°C) on the low pH (4) and a(w) (0.89) that appeared to be associated with the suppression of B. melitensis at the early stages of cheese ripening. In the ripened goat cheeses, B. melitensis survived under a precise combination of temperature during maturation, ripening time, and a(w) in the manufacturing process.

  6. Quality comparison of hydroponic tomatoes (Lycopersicon esculentum) ripened on and off vine

    NASA Technical Reports Server (NTRS)

    Arias, R.; Lee, T. C.; Specca, D.; Janes, H.

    2000-01-01

    There is a general belief that the quality of tomatoes ripened on vine is better than tomatoes ripened off the vine, influencing among other parameters, the price of this commodity. We compared the quality of hydroponic tomatoes ripened on and off vine by chemical, physical, and sensory evaluation to find what attributes are affected and to what extent. Lycopene, beta-carotene, total and soluble solids, moisture content, ascorbic acid, acidity, pH, texture, and color were analyzed. Tomatoes ripened on vine had significantly more lycopene, beta-carotene, soluble and total solids, higher a* and lower L*, and were firmer. However, a 100-judge panel rated only the color and overall liking of the vine-ripened tomatoes as more intense than the fruit ripened off vine. Therefore, the chemical and physical differences were mostly not large enough to influence the panelist's perception. The characterization of tomatoes ripened on and off vine may help to guide post-harvest handling and treatment and to improve the quality of tomatoes ripened off vine.

  7. A SHATTERPROOF-like gene controls ripening in non-climacteric strawberries, and auxin and abscisic acid antagonistically affect its expression.

    PubMed

    Daminato, Margherita; Guzzo, Flavia; Casadoro, Giorgio

    2013-09-01

    Strawberries (Fragaria×ananassa) are false fruits the ripening of which follows the non-climacteric pathway. The role played by a C-type MADS-box gene [SHATTERPROOF-like (FaSHP)] in the ripening of strawberries has been studied by transiently modifying gene expression through either over-expression or RNA-interference-mediated down-regulation. The altered expression of the FaSHP gene caused a change in the time taken by the over-expressing and the down- regulated fruits to attain the pink stage, which was slightly shorter and much longer, respectively, compared to controls. In parallel with the modified ripening times, the metabolome components and the expression of ripening-related genes also appeared different in the transiently modified fruits. Differences in the response time of the analysed genes suggest that FaSHP can control the expression of ripening genes either directly or indirectly through other transcription factor-encoding genes. Because fleshy strawberries are false fruits these results indicate that C-type MADS-box genes like SHATTERPROOF may act as modulators of ripening in fleshy fruit-like structures independently of their anatomical origin. Treatment of strawberries with either auxin or abscisic acid had antagonistic impacts on both the expression of FaSHP and the expression of ripening-related genes and metabolome components.

  8. Abscisic Acid Is a Major Regulator of Grape Berry Ripening Onset: New Insights into ABA Signaling Network

    PubMed Central

    Pilati, Stefania; Bagagli, Giorgia; Sonego, Paolo; Moretto, Marco; Brazzale, Daniele; Castorina, Giulia; Simoni, Laura; Tonelli, Chiara; Guella, Graziano; Engelen, Kristof; Galbiati, Massimo; Moser, Claudio

    2017-01-01

    Grapevine is a world-wide cultivated economically relevant crop. The process of berry ripening is non-climacteric and does not rely on the sole ethylene signal. Abscisic acid (ABA) is recognized as an important hormone of ripening inception and color development in ripening berries. In order to elucidate the effect of this signal at the molecular level, pre-véraison berries were treated ex vivo for 20 h with 0.2 mM ABA and berry skin transcriptional modulation was studied by RNA-seq after the treatment and 24 h later, in the absence of exogenous ABA. This study highlighted that a small amount of ABA triggered its own biosynthesis and had a transcriptome-wide effect (1893 modulated genes) characterized by the amplification of the transcriptional response over time. By comparing this dataset with the many studies on ripening collected within the grapevine transcriptomic compendium Vespucci, an extended overlap between ABA- and ripening modulated gene sets was observed (71% of the genes), underpinning the role of this hormone in the regulation of berry ripening. The signaling network of ABA, encompassing ABA metabolism, transport and signaling cascade, has been analyzed in detail and expanded based on knowledge from other species in order to provide an integrated molecular description of this pathway at berry ripening onset. Expression data analysis was combined with in silico promoter analysis to identify candidate target genes of ABA responsive element binding protein 2 (VvABF2), a key upstream transcription factor of the ABA signaling cascade which is up-regulated at véraison and also by ABA treatments. Two transcription factors, VvMYB143 and VvNAC17, and two genes involved in protein degradation, Armadillo-like and Xerico-like genes, were selected for in vivo validation by VvABF2-mediated promoter trans-activation in tobacco. VvNAC17 and Armadillo-like promoters were induced by ABA via VvABF2, while VvMYB143 responded to ABA in a VvABF2-independent manner. This

  9. Interactions between yeasts and bacteria in the smear surface-ripened cheeses.

    PubMed

    Corsetti, A; Rossi, J; Gobbetti, M

    2001-09-19

    In the initial phase of ripening, the microflora of bacterial smear surface-ripened cheeses such as Limburger, Taleggio, Brick, Münster and Saint-Paulin and that of surface mould-ripened cheeses such as Camembert and Brie may be similar, but at the end of the ripening, bacteria such as Brevibacterium spp., Arthrobacter spp., Micrococcus spp., Corynebacterium spp. and moulds such as Penicillium camemberti are, respectively, the dominant microorganisms. Yeasts such as Candida spp., Cryptococcus spp., Debaryomyces spp., Geotrichum candidum, Pichia spp., Rhodotorula spp., Saccharomyces spp. and Yarrowia lipolytica are often and variably isolated from the smear surface-ripened cheeses. Although not dominant within the microorganisms of the smear surface-ripened cheeses, yeasts establish significant interactions with moulds and especially bacteria, including surface bacteria and lactic acid bacteria. Some aspects of the interactions between yeasts and bacteria in such type of cheeses are considered in this paper.

  10. Characterization of Ethylene Biosynthesis Associated with Ripening in Banana Fruit1

    PubMed Central

    Liu, Xuejun; Shiomi, Shinjiro; Nakatsuka, Akira; Kubo, Yasutaka; Nakamura, Reinosuke; Inaba, Akitsugu

    1999-01-01

    We investigated the characteristics of ethylene biosynthesis associated with ripening in banana (Musa sp. [AAA group, Cavendish subgroup] cv Grand Nain) fruit. MA-ACS1 encoding 1-aminocyclopropane-1-carboxylic acid (ACC) synthase in banana fruit was the gene related to the ripening process and was inducible by exogenous ethylene. At the onset of the climacteric period in naturally ripened fruit, ethylene production increased greatly, with a sharp peak concomitant with an increase in the accumulation of MA-ACS1 mRNA, and then decreased rapidly. At the onset of ripening, the in vivo ACC oxidase activity was enhanced greatly, followed by an immediate and rapid decrease. Expression of the MA-ACO1 gene encoding banana ACC oxidase was detectable at the preclimacteric stage, increased when ripening commenced, and then remained high throughout the later ripening stage despite of a rapid reduction in the ACC oxidase activity. This discrepancy between enzyme activity and gene expression of ACC oxidase could be, at least in part, due to reduced contents of ascorbate and iron, cofactors for the enzyme, during ripening. Addition of these cofactors to the incubation medium greatly stimulated the in vivo ACC oxidase activity during late ripening stages. The results suggest that ethylene production in banana fruit is regulated by transcription of MA-ACS1 until climacteric rise and by reduction of ACC oxidase activity possibly through limited in situ availability of its cofactors once ripening has commenced, which in turn characterizes the sharp peak of ethylene production. PMID:10594112

  11. Identification of genes differentially expressed during ripening of banana.

    PubMed

    Manrique-Trujillo, Sandra Mabel; Ramírez-López, Ana Cecilia; Ibarra-Laclette, Enrique; Gómez-Lim, Miguel Angel

    2007-08-01

    The banana (Musa acuminata, subgroup Cavendish 'Grand Nain') is a climacteric fruit of economic importance. A better understanding of the banana ripening process is needed to improve fruit quality and to extend shelf life. Eighty-four up-regulated unigenes were identified by differential screening of a banana fruit cDNA subtraction library at a late ripening stage. The ripening stages in this study were defined according to the peel color index (PCI). Unigene sequences were analyzed with different databases to assign a putative identification. The expression patterns of 36 transcripts confirmed as positive by differential screening were analyzed comparing the PCI 1, PCI 5 and PCI 7 ripening stages. Expression profiles were obtained for unigenes annotated as orcinol O-methyltransferase, putative alcohol dehydrogenase, ubiquitin-protein ligase, chorismate mutase and two unigenes with non-significant matches with any reported sequence. Similar expression profiles were observed in banana pulp and peel. Our results show differential expression of a group of genes involved in processes associated with fruit ripening, such as stress, detoxification, cytoskeleton and biosynthesis of volatile compounds. Some of the identified genes had not been characterized in banana fruit. Besides providing an overview of gene expression programs and metabolic pathways at late stages of banana fruit ripening, this study contributes to increasing the information available on banana fruit ESTs.

  12. Profiling Taste and Aroma Compound Metabolism during Apricot Fruit Development and Ripening

    PubMed Central

    Xi, Wanpeng; Zheng, Huiwen; Zhang, Qiuyun; Li, Wenhui

    2016-01-01

    Sugars, organic acids and volatiles of apricot were determined by HPLC and GC-MS during fruit development and ripening, and the key taste and aroma components were identified by integrating flavor compound contents with consumers’ evaluation. Sucrose and glucose were the major sugars in apricot fruit. The contents of all sugars increased rapidly, and the accumulation pattern of sugars converted from glucose-predominated to sucrose-predominated during fruit development and ripening. Sucrose synthase (SS), sorbitol oxidase (SO) and sorbitol dehydrogenase (SDH) are under tight developmental control and they might play important roles in sugar accumulation. Almost all organic acids identified increased during early development and then decrease rapidly. During early development, fruit mainly accumulated quinate and malate, with the increase of citrate after maturation, and quinate, malate and citrate were the predominant organic acids at the ripening stage. The odor activity values (OAV) of aroma volatiles showed that 18 aroma compounds were the characteristic components of apricot fruit. Aldehydes and terpenes decreased significantly during the whole development period, whereas lactones and apocarotenoids significantly increased with fruit ripening. The partial least squares regression (PLSR) results revealed that β-ionone, γ-decalactone, sucrose and citrate are the key characteristic flavor factors contributing to consumer acceptance. Carotenoid cleavage dioxygenases (CCD) may be involved in β-ionone formation in apricot fruit. PMID:27347931

  13. Identification of potential target genes for the tomato fruit-ripening regulator RIN by chromatin immunoprecipitation.

    PubMed

    Fujisawa, Masaki; Nakano, Toshitsugu; Ito, Yasuhiro

    2011-01-30

    During ripening, climacteric fruits increase their ethylene level and subsequently undergo various physiological changes, such as softening, pigmentation and development of aroma and flavor. These changes occur simultaneously and are caused by the highly synchronized expression of numerous genes at the onset of ripening. In tomatoes, the MADS-box transcription factor RIN has been regarded as a key regulator responsible for the onset of ripening by acting upstream of both ethylene- and non-ethylene-mediated controls. However, except for LeACS2, direct targets of RIN have not been clarified, and little is known about the transcriptional cascade for ripening. Using immunoprecipitated (IPed) DNA fragments recovered by chromatin immunoprecipitation (ChIP) with anti-RIN antibody from ripening tomato fruit, we analyzed potential binding sites for RIN (CArG-box sites) in the promoters of representative ripening-induced genes by quantitative PCR. Results revealed nearly a 5- to 20-fold enrichment of CArG boxes in the promoters of LeACS2, LeACS4, PG, TBG4, LeEXP1, and LeMAN4 and of RIN itself, indicating direct interaction of RIN with their promoters in vivo. Moreover, sequence analysis and genome mapping of 51 cloned IPed DNAs revealed potential RIN binding sites. Quantitative PCR revealed that four of the potential binding sites were enriched 4- to 17-fold in the IPed DNA pools compared with the controls, indicating direct interaction of RIN with these sites in vivo. Near one of the four CArG boxes we found a gene encoding a protein similar to thioredoxin y1. An increase in the transcript level of this gene was observed with ripening in normal fruit but not in the rin mutant, suggesting that RIN possibly induces its expression. The presented results suggest that RIN controls fruit softening and ethylene production by the direct transcriptional regulation of cell-wall-modifying genes and ethylene biosynthesis genes during ripening. Moreover, the binding of RIN to its own

  14. Home cervical ripening with dinoprostone gel in nulliparous women with singleton pregnancies.

    PubMed

    Stock, Sarah J; Taylor, Rebecca; Mairs, Rebecca; Azaghdani, Abdulhamid; Hor, Kahyee; Smith, Imogen; Dundas, Kirsty; Kissack, Chris; Norman, Jane E; Denison, Fiona

    2014-08-01

    To evaluate whether home cervical ripening is safe and results in shorter hospital stay. This was a retrospective cohort study of women with singleton pregnancies having induction of labor for postmaturity at a single center between January 2007 and June 2010. Women were offered home cervical ripening with 1 mg dinoprostone gel if they were nulliparous, had uncomplicated singleton pregnancies, and the indication for induction was postmaturity. Nine hundred seven of 1,536 (59.1%) nulliparous women having induction of labor for postmaturity were eligible for home cervical ripening. The median number of hours at home was 11.76 hours (range 0-24.82 hours). There were no cases of birth outside of the hospital, uterine rupture, or significant neonatal morbidity or neonatal death related to home cervical ripening. Eighty-five (5.5%) women who underwent hospital cervical ripening because of maternal preference or social issues formed a hospital cervical ripening comparison group. There was no significant difference in the total number of hours before delivery spent in the hospital between the two groups (26.25; 95% confidence interval [CI] 25.27-27.23 in home cervical ripening group compared with 24.28; 95% CI 22.5-26.0 in the hospital group; P=.26). Clinical outcomes are comparable in nulliparous women who receive a single dose of dinoprostone gel for home cervical ripening compared with those who undergo hospital cervical ripening. However, preadmission home cervical ripening with 1 mg dinoprostone does not decrease the number of hours women spend in the hospital. II.

  15. Inhibitors of Ethylene Biosynthesis and Signaling.

    PubMed

    Schaller, G Eric; Binder, Brad M

    2017-01-01

    Ethylene is a gas biosynthesized by plants which has many physiological and developmental effects on their growth. Ethylene affects agriculturally and horticulturally important traits such as fruit ripening, post-harvest physiology, senescence, and abscission, and so ethylene action is often inhibited to improve the shelf life of fruits, vegetables, and cut flowers. Chemical inhibitors of ethylene action are also useful for research to characterize the mechanisms of ethylene biosynthesis and signal transduction, and the role that ethylene plays in various physiological processes. Here, we describe the use of three inhibitors commonly used for the study of ethylene action in plants: 2-aminoethoxyvinyl glycine (AVG), silver ions (Ag), and the gaseous compound 1-methylcyclopropene (1-MCP). AVG is an inhibitor of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, a key enzyme involved in ethylene biosynthesis. Silver and 1-MCP are both inhibitors of the ethylene receptors. Inhibitor use as well as off-target effects are described with a focus on ethylene responses in dark-grown Arabidopsis seedlings. Methods for the use of these inhibitors can be applied to other plant growth assays.

  16. Proteomics as an approach to the understanding of the molecular physiology of fruit development and ripening.

    PubMed

    Palma, José M; Corpas, Francisco J; del Río, Luís A

    2011-08-12

    Fruit ripening is a developmental complex process which occurs in higher plants and involves a number of stages displayed from immature to mature fruits that depend on the plant species and the environmental conditions. Nowadays, the importance of fruit ripening comes mainly from the link between this physiological process in plants and the economic repercussions as a result of one of the human activities, the agricultural industry. In most cases, fruit ripening is accompanied by colour changes due to different pigment content and increases in sugar levels, among others. Major physiological modifications that affect colour, texture, flavour, and aroma are under the control of both external (light and temperature) and internal (developmental gene regulation and hormonal control) factors. Due to the huge amount of metabolic changes that take place during ripening in fruits from higher plants, the accomplishment of new throughput methods which can provide a global evaluation of this process would be desirable. Differential proteomics of immature and mature fruits would be a useful tool to gain information on the molecular changes which occur during ripening, but also the investigation of fruits at different ripening stages will provide a dynamic picture of the whole transformation of fruits. This subject is furthermore of great interest as many fruits are essential for human nutrition. Thus far different maturation profiles have been reported specific for each crop species. In this work, a thorough review of the proteomic database from fruit development and maturation of important crop species will be updated to understand the molecular physiology of fruits at ripening stages. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Microarray analysis of gene expression profiles in ripening pineapple fruits.

    PubMed

    Koia, Jonni H; Moyle, Richard L; Botella, Jose R

    2012-12-18

    Pineapple (Ananas comosus) is a tropical fruit crop of significant commercial importance. Although the physiological changes that occur during pineapple fruit development have been well characterized, little is known about the molecular events that occur during the fruit ripening process. Understanding the molecular basis of pineapple fruit ripening will aid the development of new varieties via molecular breeding or genetic modification. In this study we developed a 9277 element pineapple microarray and used it to profile gene expression changes that occur during pineapple fruit ripening. Microarray analyses identified 271 unique cDNAs differentially expressed at least 1.5-fold between the mature green and mature yellow stages of pineapple fruit ripening. Among these 271 sequences, 184 share significant homology with genes encoding proteins of known function, 53 share homology with genes encoding proteins of unknown function and 34 share no significant homology with any database accession. Of the 237 pineapple sequences with homologs, 160 were up-regulated and 77 were down-regulated during pineapple fruit ripening. DAVID Functional Annotation Cluster (FAC) analysis of all 237 sequences with homologs revealed confident enrichment scores for redox activity, organic acid metabolism, metalloenzyme activity, glycolysis, vitamin C biosynthesis, antioxidant activity and cysteine peptidase activity, indicating the functional significance and importance of these processes and pathways during pineapple fruit development. Quantitative real-time PCR analysis validated the microarray expression results for nine out of ten genes tested. This is the first report of a microarray based gene expression study undertaken in pineapple. Our bioinformatic analyses of the transcript profiles have identified a number of genes, processes and pathways with putative involvement in the pineapple fruit ripening process. This study extends our knowledge of the molecular basis of pineapple fruit

  18. Microarray analysis of gene expression profiles in ripening pineapple fruits

    PubMed Central

    2012-01-01

    Background Pineapple (Ananas comosus) is a tropical fruit crop of significant commercial importance. Although the physiological changes that occur during pineapple fruit development have been well characterized, little is known about the molecular events that occur during the fruit ripening process. Understanding the molecular basis of pineapple fruit ripening will aid the development of new varieties via molecular breeding or genetic modification. In this study we developed a 9277 element pineapple microarray and used it to profile gene expression changes that occur during pineapple fruit ripening. Results Microarray analyses identified 271 unique cDNAs differentially expressed at least 1.5-fold between the mature green and mature yellow stages of pineapple fruit ripening. Among these 271 sequences, 184 share significant homology with genes encoding proteins of known function, 53 share homology with genes encoding proteins of unknown function and 34 share no significant homology with any database accession. Of the 237 pineapple sequences with homologs, 160 were up-regulated and 77 were down-regulated during pineapple fruit ripening. DAVID Functional Annotation Cluster (FAC) analysis of all 237 sequences with homologs revealed confident enrichment scores for redox activity, organic acid metabolism, metalloenzyme activity, glycolysis, vitamin C biosynthesis, antioxidant activity and cysteine peptidase activity, indicating the functional significance and importance of these processes and pathways during pineapple fruit development. Quantitative real-time PCR analysis validated the microarray expression results for nine out of ten genes tested. Conclusions This is the first report of a microarray based gene expression study undertaken in pineapple. Our bioinformatic analyses of the transcript profiles have identified a number of genes, processes and pathways with putative involvement in the pineapple fruit ripening process. This study extends our knowledge of the

  19. Detection of artificially ripened mango using spectrometric analysis

    NASA Astrophysics Data System (ADS)

    Mithun, B. S.; Mondal, Milton; Vishwakarma, Harsh; Shinde, Sujit; Kimbahune, Sanjay

    2017-05-01

    Hyperspectral sensing has been proven to be useful to determine the quality of food in general. It has also been used to distinguish naturally and artificially ripened mangoes by analyzing the spectral signature. However the focus has been on improving the accuracy of classification after performing dimensionality reduction, optimum feature selection and using suitable learning algorithm on the complete visible and NIR spectrum range data, namely 350nm to 1050nm. In this paper we focus on, (i) the use of low wavelength resolution and low cost multispectral sensor to reliably identify artificially ripened mango by selectively using the spectral information so that classification accuracy is not hampered at the cost of low resolution spectral data and (ii) use of visible spectrum i.e. 390nm to 700 nm data to accurately discriminate artificially ripened mangoes. Our results show that on a low resolution spectral data, the use of logistic regression produces an accuracy of 98.83% and outperforms other methods like classification tree, random forest significantly. And this is achieved by analyzing only 36 spectral reflectance data points instead of the complete 216 data points available in visual and NIR range. Another interesting experimental observation is that we are able to achieve more than 98% classification accuracy by selecting only 15 irradiance values in the visible spectrum. Even the number of data needs to be collected using hyper-spectral or multi-spectral sensor can be reduced by a factor of 24 for classification with high degree of confidence

  20. Distribution of phenolic compounds and antioxidant capacity in apples tissues during ripening.

    PubMed

    Alberti, Aline; Zielinski, Acácio Antonio Ferreira; Couto, Marcelo; Judacewski, Priscila; Mafra, Luciana Igarashi; Nogueira, Alessandro

    2017-05-01

    The effect of variety and ripening stage on the distribution of phenolic compounds and in vitro antioxidant capacity of Gala, Fuji Suprema and Eva apples were evaluated. Hydroxycinnamic acids, flavonoids, flavanols, flavonols, dihydrochalcones and antioxidant activity (FRAP and DPPH) were assessed in the epicarp, mesocarp and endocarp of three varieties at three ripening stages (unripe, ripe and senescent). The Fuji Suprema variety distinguished by its content of flavonols at senescent stage, while Eva variety distinguished by its content of dihydrochalcones (unripe stage) and anthocyanins (ripe stage). In general, phenolic acids and flavonoids decreased with ripening in the epicarp and endocarp. However, in the mesocarp, the effect of ripening was related with the apple variety. Hierarchical cluster analysis confirmed the influence of ripening in the apple tissue. The evolution of these compounds during ripening occurred irregularly and it was influenced by the variety.

  1. Ethylene-producing bacteria that ripen fruit.

    PubMed

    Digiacomo, Fabio; Girelli, Gabriele; Aor, Bruno; Marchioretti, Caterina; Pedrotti, Michele; Perli, Thomas; Tonon, Emil; Valentini, Viola; Avi, Damiano; Ferrentino, Giovanna; Dorigato, Andrea; Torre, Paola; Jousson, Olivier; Mansy, Sheref S; Del Bianco, Cristina

    2014-12-19

    Ethylene is a plant hormone widely used to ripen fruit. However, the synthesis, handling, and storage of ethylene are environmentally harmful and dangerous. We engineered E. coli to produce ethylene through the activity of the ethylene-forming enzyme (EFE) from Pseudomonas syringae. EFE converts a citric acid cycle intermediate, 2-oxoglutarate, to ethylene in a single step. The production of ethylene was placed under the control of arabinose and blue light responsive regulatory systems. The resulting bacteria were capable of accelerating the ripening of tomatoes, kiwifruit, and apples.

  2. Fruit ripening using hyperspectral imaging

    NASA Astrophysics Data System (ADS)

    ., Swetha; Chidangil, Santhosh; Karpate, Tanvi; Asundi, Anand

    2017-06-01

    The ripening of fruits is associated with changes, in some cases subtle, in the color of the fruit. Traditionally spectroscopy used to measure these subtle changes and infer the ripeness of fruits. Spectrometers provides high-resolution but only measure a small area of the fruit. That might not be a good indicator of the overall ripeness. In this paper, we propose a compact tunable LED based hyper spectral imaging system that scans through a set of wavelengths and images, the reflectance from the whole fruit. Based on the type of fruit, only specific wavelengths need to be scanned. Following a validation using a Rubik's cube, an example banana going through its ripening cycles is used to demonstrate the system.

  3. Transcriptomic analysis of apple fruit ripening and texture attributes

    USDA-ARS?s Scientific Manuscript database

    Molecular events regulating cultivar-specific apple fruit ripening and sensory quality are largely unknown. Such knowledge is essential for genomic-assisted apple breeding and postharvest quality management. The ripening behavior and texture attributes of two apple cultivars, ‘Pink Lady’ and ‘Honey...

  4. The effect of application of cold natural smoke on the ripening of Cheddar cheese.

    PubMed

    Shakeel-Ur-Rehman; Farkye, N Y; Drake, M A

    2003-06-01

    The present study was undertaken to study the effects of application of natural wood smoke on ripening of Cheddar cheese, and to determine the effects of smoking before or after ripening on cheese quality. A 20-kg block of Cheddar cheese obtained immediately after pressing was divided into six approximately 3-kg blocks and ripened at 8 degrees C for up to 270 d. One 3-kg block was taken after 1 d, 1, 3, 6, or 9 mo and smoked for 20 min, then returned to the ripening room for further ripening. Cheeses were sampled at intervals for lactobacilli counts, moisture, pH, and proteolysis. Sensory analysis was conducted on 6 and 9-mo-old cheeses by a trained sensory panel (n = 7). Results show that application of natural wood smoke did not significantly affect cheese pH or primary proteolysis during ripening. However, secondary proteolysis as assessed by the concentrations of free amino acids was generally higher in smoked cheeses than in control cheeses after 6 mo of ripening. Cheese smoked after 6 mo of ripening had better smoked flavor than that smoked after 9 mo of ripening. Cheese smoked after 3 mo of age and further ripened for 6 mo had the highest smoked flavor intensity. It is concluded that it is best to smoke cheese after ripening for at least 3 mo.

  5. Effect of the potassium permanganate during papaya fruit ripening: Ethylene production

    NASA Astrophysics Data System (ADS)

    Corrêa, S. F.; Filho, M. B.; da Silva, M. G.; Oliveira, J. G.; Aroucha, E. M. M.; Silva, R. F.; Pereira, M. G.; Vargas, H.

    2005-06-01

    The effect of potassium permanganate (KMnO4) on the ripening process of papaya fruits by monitoring the ethylene emission rates is reported. The ethylene emission was monitored by a photoacoustic spectrometer. Two experimental conditions were applied, being one of them just putting the fruit alone inside the sampling chamber and the second, modifying the atmosphere by the presence of KMnO4. The use of the ethylene absorber reduces the autocatalytic process of ethylene during papaya fruit ripening. For 20 g of KMnO4 the maximal intensity of the ethylene emission decreases by a factor two. Using the same amount of KMnO4, a reduction of about 2.2% in the concentration of ethylene for a mixture of 1ppmv of ethylene in synthetic air was observed.

  6. Red blotch disease alters grape berry development and metabolism by interfering with the transcriptional and hormonal regulation of ripening

    PubMed Central

    Blanco-Ulate, Barbara; Hopfer, Helene; Figueroa-Balderas, Rosa; Ye, Zirou; Rivero, Rosa M.; Albacete, Alfonso; Pérez-Alfocea, Francisco; Koyama, Renata; Anderson, Michael M.; Smith, Rhonda J.; Ebeler, Susan E.

    2017-01-01

    Abstract Grapevine red blotch-associated virus (GRBaV) is a major threat to the wine industry in the USA. GRBaV infections (aka red blotch disease) compromise crop yield and berry chemical composition, affecting the flavor and aroma properties of must and wine. In this study, we combined genome-wide transcriptional profiling with targeted metabolite analyses and biochemical assays to characterize the impact of the disease on red-skinned berry ripening and metabolism. Using naturally infected berries collected from two vineyards, we were able to identify consistent berry responses to GRBaV across different environmental and cultural conditions. Specific alterations of both primary and secondary metabolism occurred in GRBaV-infected berries during ripening. Notably, GRBaV infections of post-véraison berries resulted in the induction of primary metabolic pathways normally associated with early berry development (e.g. thylakoid electron transfer and the Calvin cycle), while inhibiting ripening-associated pathways, such as a reduced metabolic flux in the central and peripheral phenylpropanoid pathways. We show that this metabolic reprogramming correlates with perturbations at multiple regulatory levels of berry development. Red blotch caused the abnormal expression of transcription factors (e.g. NACs, MYBs, and AP2-ERFs) and elements of the post-transcriptional machinery that function during red-skinned berry ripening. Abscisic acid, ethylene, and auxin pathways, which control both the initiation of ripening and stress responses, were also compromised. We conclude that GRBaV infections disrupt normal berry development and stress responses by altering transcription factors and hormone networks, which result in the inhibition of ripening pathways involved in the generation of color, flavor, and aroma compounds. PMID:28338755

  7. Short communication: Effect of genetic type on antioxidant activity of Caciocavallo cheese during ripening.

    PubMed

    Perna, Annamaria; Intaglietta, Immacolata; Simonetti, Amalia; Gambacorta, Emilio

    2015-06-01

    The aim of this work was to investigate the antioxidant activity of Caciocavallo cheese made from the milk of 2 breeds, Italian Brown and Italian Holstein, and ripened for 1, 30, 60, 90, and 150 d. The antioxidant activity of cheese was measured using the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), ferric-reducing antioxidant power (FRAP), and thiol assays. Statistical analysis showed a significant effect of the studied factors. Italian Brown cheese had higher antioxidant activity than Italian Holstein cheese, and antioxidant activity increased during ripening of both cheeses types. Moreover, antioxidant activity varied during ripening depending on the rate of formation of soluble peptides. To date, few studies have evaluated the effect of genetic type on antioxidant capacity of the pasta filata cheeses; thus, this study forms the basis of new knowledge that could lead to the production of a pasta filata cheese with specific nutraceutical characteristics. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  8. Transcriptional transitions in Alphonso mango (Mangifera indica L.) during fruit development and ripening explain its distinct aroma and shelf life characteristics.

    PubMed

    Deshpande, Ashish B; Anamika, Krishanpal; Jha, Vineet; Chidley, Hemangi G; Oak, Pranjali S; Kadoo, Narendra Y; Pujari, Keshav H; Giri, Ashok P; Gupta, Vidya S

    2017-08-18

    Alphonso is known as the "King of mangos" due to its unique flavor, attractive color, low fiber pulp and long shelf life. We analyzed the transcriptome of Alphonso mango through Illumina sequencing from seven stages of fruit development and ripening as well as flower. Total transcriptome data from these stages ranged between 65 and 143 Mb. Importantly, 20,755 unique transcripts were annotated and 4,611 were assigned enzyme commission numbers, which encoded 142 biological pathways. These included ethylene and flavor related secondary metabolite biosynthesis pathways, as well as those involved in metabolism of starch, sucrose, amino acids and fatty acids. Differential regulation (p-value ≤ 0.05) of thousands of transcripts was evident in various stages of fruit development and ripening. Novel transcripts for biosynthesis of mono-terpenes, sesqui-terpenes, di-terpenes, lactones and furanones involved in flavor formation were identified. Large number of transcripts encoding cell wall modifying enzymes was found to be steady in their expression, while few were differentially regulated through these stages. Novel 79 transcripts of inhibitors of cell wall modifying enzymes were simultaneously detected throughout Alphonso fruit development and ripening, suggesting controlled activity of these enzymes involved in fruit softening.

  9. A DEMETER-like DNA demethylase governs tomato fruit ripening

    USDA-ARS?s Scientific Manuscript database

    This work shows that active DNA demethylation governs ripening, an important plant developmental process. Our work defines a molecular mechanism, which has until now been missing, to explain the correlation between genomic DNA demethylation and fruit ripening. It demonstrates a direct cause-and-effe...

  10. Effect of Gamma Radiation on the Ripening of Bartlett Pears 1

    PubMed Central

    Maxie, E. C.; Sommer, N. F.; Muller, Carlos J.; Rae, Henry L.

    1966-01-01

    Gamma radiation at doses of 300 Krad or more inhibits the ripening of Bartlett pears (Pyrus communis L.). Immediately after irradiation there is a transitory burst of C2H4, which subsequently declines in fruits subjected to inhibitory doses. Ethylene production associated with ripening begins at the same time in unirradiated fruits and those subjected to noninhibitory doses, but the latter produces much more C2H4 at the climacteric peak. Fruits subjected to inhibitory doses produce low levels of C2H4 unless subjected to exogenously applied C2H4, whereupon they produce enough of the gas to induce ripening in unirradiated fruits. Pears subjected to 300 and 400 Krad of gamma rays did not ripen even when held in a flowing atmosphere containing 1000 ppm of C2H4 for 8 days at 20°. It is concluded that the action of gamma rays on Bartlett pears involves both an inhibition of C2H4 production and a decreased sensitivity of the fruit to the ripening action of the gas. Ripening of Bartlett pears is inhibited by gamma radiation only when applied to preclimacteric fruit. PMID:16656274

  11. Metabolite Profiling Reveals Developmental Inequalities in Pinot Noir Berry Tissues Late in Ripening.

    PubMed

    Vondras, Amanda M; Commisso, Mauro; Guzzo, Flavia; Deluc, Laurent G

    2017-01-01

    Uneven ripening in Vitis vinifera is increasingly recognized as a phenomenon of interest, with substantial implications for fruit and wine composition and quality. This study sought to determine whether variation late in ripening (∼Modified Eichhorn-Lorenz stage 39) was associated with developmental differences that were observable as fruits within a cluster initiated ripening (véraison). Four developmentally distinct ripening classes of berries were tagged at cluster véraison, sampled at three times late in ripening, and subjected to untargeted HPLC-MS to measure variation in amino acids, sugars, organic acids, and phenolic metabolites in skin, pulp, and seed tissues separately. Variability was described using predominantly two strategies. In the first, multivariate analysis (Orthogonal Projections to Latent Structures-Discriminant Analysis, OPLS-DA) was used to determine whether fruits were still distinguishable per their developmental position at véraison and to identify which metabolites accounted for these distinctions. The same technique was used to assess changes in each tissue over time. In a second strategy and for each annotated metabolite, the variance across the ripening classes at each time point was measured to show whether intra-cluster variance (ICV) was growing, shrinking, or constant over the period observed. Indeed, berries could be segregated by OPLS-DA late in ripening based on their developmental position at véraison, though the four ripening classes were aggregated into two larger ripening groups. Further, not all tissues were dynamic over the period examined. Although pulp tissues could be segregated by time sampled, this was not true for seed and only moderately so for skin. Ripening group differences in seed and skin, rather than the time fruit was sampled, were better able to define berries. Metabolites also experienced significant reductions in ICV between single pairs of time points, but never across the entire experiment

  12. Physiological, molecular and ultrastructural analyses during ripening and over-ripening of banana (Musa spp., AAA group, Cavendish sub-group) fruit suggest characteristics of programmed cell death.

    PubMed

    Ramírez-Sánchez, Maricruz; Huber, Donald J; Vallejos, C Eduardo; Kelley, Karen

    2018-01-01

    Programmed cell death (PCD) is a part of plant development that has been studied for petal senescence and vegetative tissue but has not been thoroughly investigated for fleshy fruits. The purpose of this research was to examine ripening and over-ripening in banana fruit to determine if there were processes in common to previously described PCD. Loss of cellular integrity (over 40%) and development of senescence related dark spot (SRDS) occurred after day 8 in banana peel. Nuclease and protease activity in the peel increased during ripening starting from day 2, and decreased during over-ripening. The highest activity was for proteases and nucleases with apparent molecular weights of 86 kDa and 27 kDa, respectively. Images of SRDS showed shrinkage of the upper layers of cells, visually suggesting cell death. Decrease of electron dense areas was evident in TEM micrographs of nuclei. This study shows for the first time that ripening and over-ripening of banana peel share physiological and molecular processes previously described in plant PCD. SRDS could represent a morphotype of PCD that characterizes a structural and biochemical failure in the upper layers of the peel, thereafter spreading to lower and adjacent layers of cells. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  13. Early stages of Ostwald ripening

    NASA Astrophysics Data System (ADS)

    Shneidman, Vitaly A.

    2013-07-01

    The Becker-Döring (BD) nucleation equation is known to predict a narrow double-exponential front (DEF) in the distribution of growing particles over sizes, which is due to early transient effects. When mass conservation is included, nucleation is eventually exhausted while independent growth is replaced by ripening. Despite the enormous difference in the associated time scales, and the resulting demand on numerics, within the generalized BD model the early DEF is shown to be crucial for the selection of the unique self-similar Lifshitz-Slyozov-Wagner asymptotic regime. Being preserved till the latest stages of growth, the DEF provides a universal part of the initial conditions for the ripening problem, regardless of the mass exchange mechanism between the nucleus and the matrix.

  14. Tomato fruit carotenoid biosynthesis is adjusted to actual ripening progression by a light-dependent mechanism.

    PubMed

    Llorente, Briardo; D'Andrea, Lucio; Ruiz-Sola, M Aguila; Botterweg, Esther; Pulido, Pablo; Andilla, Jordi; Loza-Alvarez, Pablo; Rodriguez-Concepcion, Manuel

    2016-01-01

    Carotenoids are isoprenoid compounds that are essential for plants to protect the photosynthetic apparatus against excess light. They also function as health-promoting natural pigments that provide colors to ripe fruit, promoting seed dispersal by animals. Work in Arabidopsis thaliana unveiled that transcription factors of the phytochrome-interacting factor (PIF) family regulate carotenoid gene expression in response to environmental signals (i.e. light and temperature), including those created when sunlight reflects from or passes though nearby vegetation or canopy (referred to as shade). Here we show that PIFs use a virtually identical mechanism to modulate carotenoid biosynthesis during fruit ripening in tomato (Solanum lycopersicum). However, instead of integrating environmental information, PIF-mediated signaling pathways appear to fulfill a completely new function in the fruit. As tomatoes ripen, they turn from green to red due to chlorophyll breakdown and carotenoid accumulation. When sunlight passes through the flesh of green fruit, a self-shading effect within the tissue maintains high levels of PIFs that directly repress the master gene of the fruit carotenoid pathway, preventing undue production of carotenoids. This effect is attenuated as chlorophyll degrades, causing degradation of PIF proteins and boosting carotenoid biosynthesis as ripening progresses. Thus, shade signaling components may have been co-opted in tomato fruit to provide information on the actual stage of ripening (based on the pigment profile of the fruit at each moment) and thus finely coordinate fruit color change. We show how this mechanism may be manipulated to obtain carotenoid-enriched fruits. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

  15. iTRAQ-Based Quantitative Proteomics of Developing and Ripening Muscadine Grape Berry

    PubMed Central

    Kambiranda, Devaiah; Katam, Ramesh; Basha, Sheikh M.; Siebert, Shalom

    2014-01-01

    Grapes are among the widely cultivated fruit crops in the world. Grape berries like other nonclimacteric fruits undergo a complex set of dynamic, physical, physiological, and biochemical changes during ripening. Muscadine grapes are widely cultivated in the southern United States for fresh fruit and wine. To date, changes in the metabolites composition of muscadine grapes have been well documented; however, the molecular changes during berry development and ripening are not fully known. The aim of this study was to investigate changes in the berry proteome during ripening in muscadine grape cv. Noble. Isobaric tags for relative and absolute quantification (iTRAQ) MS/MS was used to detect statistically significant changes in the berry proteome. A total of 674 proteins were detected, and 76 were differentially expressed across four time points in muscadine berry. Proteins obtained were further analyzed to provide information about its potential functions during ripening. Several proteins involved in abiotic and biotic stimuli and sucrose and hexose metabolism were upregulated during berry ripening. Quantitative real-time PCR analysis validated the protein expression results for nine proteins. Identification of vicilin-like antimicrobial peptides indicates additional disease tolerance proteins are present in muscadines for berry protection during ripening. The results provide new information for characterization and understanding muscadine berry proteome and grape ripening. PMID:24251720

  16. Red blotch disease alters grape berry development and metabolism by interfering with the transcriptional and hormonal regulation of ripening.

    PubMed

    Blanco-Ulate, Barbara; Hopfer, Helene; Figueroa-Balderas, Rosa; Ye, Zirou; Rivero, Rosa M; Albacete, Alfonso; Pérez-Alfocea, Francisco; Koyama, Renata; Anderson, Michael M; Smith, Rhonda J; Ebeler, Susan E; Cantu, Dario

    2017-02-01

    Grapevine red blotch-associated virus (GRBaV) is a major threat to the wine industry in the USA. GRBaV infections (aka red blotch disease) compromise crop yield and berry chemical composition, affecting the flavor and aroma properties of must and wine. In this study, we combined genome-wide transcriptional profiling with targeted metabolite analyses and biochemical assays to characterize the impact of the disease on red-skinned berry ripening and metabolism. Using naturally infected berries collected from two vineyards, we were able to identify consistent berry responses to GRBaV across different environmental and cultural conditions. Specific alterations of both primary and secondary metabolism occurred in GRBaV-infected berries during ripening. Notably, GRBaV infections of post-véraison berries resulted in the induction of primary metabolic pathways normally associated with early berry development (e.g. thylakoid electron transfer and the Calvin cycle), while inhibiting ripening-associated pathways, such as a reduced metabolic flux in the central and peripheral phenylpropanoid pathways. We show that this metabolic reprogramming correlates with perturbations at multiple regulatory levels of berry development. Red blotch caused the abnormal expression of transcription factors (e.g. NACs, MYBs, and AP2-ERFs) and elements of the post-transcriptional machinery that function during red-skinned berry ripening. Abscisic acid, ethylene, and auxin pathways, which control both the initiation of ripening and stress responses, were also compromised. We conclude that GRBaV infections disrupt normal berry development and stress responses by altering transcription factors and hormone networks, which result in the inhibition of ripening pathways involved in the generation of color, flavor, and aroma compounds. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  17. 7 CFR 932.109 - Canned ripe olives of the tree-ripened type.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Canned ripe olives of the tree-ripened type. 932.109... OLIVES GROWN IN CALIFORNIA Rules and Regulations § 932.109 Canned ripe olives of the tree-ripened type. (a) Canned ripe olives of the tree-ripened type means packaged olives, not oxidized in processing...

  18. 7 CFR 932.109 - Canned ripe olives of the tree-ripened type.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 8 2014-01-01 2014-01-01 false Canned ripe olives of the tree-ripened type. 932.109... OLIVES GROWN IN CALIFORNIA Rules and Regulations § 932.109 Canned ripe olives of the tree-ripened type. (a) Canned ripe olives of the tree-ripened type means packaged olives, not oxidized in processing...

  19. 7 CFR 932.109 - Canned ripe olives of the tree-ripened type.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 8 2012-01-01 2012-01-01 false Canned ripe olives of the tree-ripened type. 932.109... OLIVES GROWN IN CALIFORNIA Rules and Regulations § 932.109 Canned ripe olives of the tree-ripened type. (a) Canned ripe olives of the tree-ripened type means packaged olives, not oxidized in processing...

  20. 7 CFR 932.109 - Canned ripe olives of the tree-ripened type.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 8 2011-01-01 2011-01-01 false Canned ripe olives of the tree-ripened type. 932.109... OLIVES GROWN IN CALIFORNIA Rules and Regulations § 932.109 Canned ripe olives of the tree-ripened type. (a) Canned ripe olives of the tree-ripened type means packaged olives, not oxidized in processing...

  1. 7 CFR 932.109 - Canned ripe olives of the tree-ripened type.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 8 2013-01-01 2013-01-01 false Canned ripe olives of the tree-ripened type. 932.109... OLIVES GROWN IN CALIFORNIA Rules and Regulations § 932.109 Canned ripe olives of the tree-ripened type. (a) Canned ripe olives of the tree-ripened type means packaged olives, not oxidized in processing...

  2. Search for Transcriptional and Metabolic Markers of Grape Pre-Ripening and Ripening and Insights into Specific Aroma Development in Three Portuguese Cultivars

    PubMed Central

    Sousa, Lisete; Pais, Maria Salomé; Kopka, Joachim; Fortes, Ana Margarida

    2013-01-01

    Background Grapes (Vitis species) are economically the most important fruit crop worldwide. However, the complexity of molecular and biochemical events that lead to ripening of berries as well as how aroma is developed are not fully understood. Methodology/Principal Findings In an attempt to identify the common mechanisms associated with the onset of ripening independently of the cultivar, grapes of Portuguese elite cultivars, Trincadeira, Aragonês, and Touriga Nacional, were studied. The mRNA expression profiles corresponding to veraison (EL35) and mature berries (EL36) were compared. Across the three varieties, 9,8% (2255) probesets corresponding to 1915 unigenes were robustly differentially expressed at EL 36 compared to EL 35. Eleven functional categories were represented in this differential gene set. Information on gene expression related to primary and secondary metabolism was verified by RT-qPCR analysis of selected candidate genes at four developmental stages (EL32, EL35, EL36 and EL 38). Gene expression data were integrated with metabolic profiling data from GC-EI-TOF/MS and headspace GC-EI-MS platforms. Conclusions/Significance Putative molecular and metabolic markers of grape pre-ripening and ripening related to primary and secondary metabolism were established and revealed a substantial developmental reprogramming of cellular metabolism. Altogether the results provide valuable new information on the main metabolic events leading to grape ripening. Furthermore, we provide first hints about how the development of a cultivar specific aroma is controlled at transcriptional level. PMID:23565246

  3. Identification, cloning and characterization of the tomato TCP transcription factor family.

    PubMed

    Parapunova, Violeta; Busscher, Marco; Busscher-Lange, Jacqueline; Lammers, Michiel; Karlova, Rumyana; Bovy, Arnaud G; Angenent, Gerco C; de Maagd, Ruud A

    2014-06-06

    TCP proteins are plant-specific transcription factors, which are known to have a wide range of functions in different plant species such as in leaf development, flower symmetry, shoot branching, and senescence. Only a small number of TCP genes has been characterised from tomato (Solanum lycopersicum). Here we report several functional features of the members of the entire family present in the tomato genome. We have identified 30 Solanum lycopersicum SlTCP genes, most of which have not been described before. Phylogenetic analysis clearly distinguishes two homology classes of the SlTCP transcription factor family - class I and class II. Class II differentiates in two subclasses, the CIN-TCP subclass and the CYC/TB1 subclass, involved in leaf development and axillary shoots formation, respectively. The expression patterns of all members were determined by quantitative PCR. Several SlTCP genes, like SlTCP12, SlTCP15 and SlTCP18 are preferentially expressed in the tomato fruit, suggesting a role during fruit development or ripening. These genes are regulated by RIN (RIPENING INHIBITOR), CNR (COLORLESS NON-RIPENING) and SlAP2a (APETALA2a) proteins, which are transcription factors with key roles in ripening. With a yeast one-hybrid assay we demonstrated that RIN binds the promoter fragments of SlTCP12, SlTCP15 and SlTCP18, and that CNR binds the SlTCP18 promoter. This data strongly suggests that these class I SlTCP proteins are involved in ripening. Furthermore, we demonstrate that SlTCPs bind the promoter fragments of members of their own family, indicating that they regulate each other. Additional yeast one-hybrid studies performed with Arabidopsis transcription factors revealed binding of the promoter fragments by proteins involved in the ethylene signal transduction pathway, contributing to the idea that these SlTCP genes are involved in the ripening process. Yeast two-hybrid data shows that SlTCP proteins can form homo and heterodimers, suggesting that they act

  4. Identification, cloning and characterization of the tomato TCP transcription factor family

    PubMed Central

    2014-01-01

    Background TCP proteins are plant-specific transcription factors, which are known to have a wide range of functions in different plant species such as in leaf development, flower symmetry, shoot branching, and senescence. Only a small number of TCP genes has been characterised from tomato (Solanum lycopersicum). Here we report several functional features of the members of the entire family present in the tomato genome. Results We have identified 30 Solanum lycopersicum SlTCP genes, most of which have not been described before. Phylogenetic analysis clearly distinguishes two homology classes of the SlTCP transcription factor family - class I and class II. Class II differentiates in two subclasses, the CIN-TCP subclass and the CYC/TB1 subclass, involved in leaf development and axillary shoots formation, respectively. The expression patterns of all members were determined by quantitative PCR. Several SlTCP genes, like SlTCP12, SlTCP15 and SlTCP18 are preferentially expressed in the tomato fruit, suggesting a role during fruit development or ripening. These genes are regulated by RIN (RIPENING INHIBITOR), CNR (COLORLESS NON-RIPENING) and SlAP2a (APETALA2a) proteins, which are transcription factors with key roles in ripening. With a yeast one-hybrid assay we demonstrated that RIN binds the promoter fragments of SlTCP12, SlTCP15 and SlTCP18, and that CNR binds the SlTCP18 promoter. This data strongly suggests that these class I SlTCP proteins are involved in ripening. Furthermore, we demonstrate that SlTCPs bind the promoter fragments of members of their own family, indicating that they regulate each other. Additional yeast one-hybrid studies performed with Arabidopsis transcription factors revealed binding of the promoter fragments by proteins involved in the ethylene signal transduction pathway, contributing to the idea that these SlTCP genes are involved in the ripening process. Yeast two-hybrid data shows that SlTCP proteins can form homo and heterodimers, suggesting

  5. Ostwald ripening of clays and metamorphic minerals

    USGS Publications Warehouse

    Eberl, D.D.; Srodon, J.; Kralik, M.; Taylor, B.E.; Peterman, Z.E.

    1990-01-01

    Analyses of particle size distributions indicate that clay minerals and other diagenetic and metamorphic minerals commonly undergo recrystallization by Ostwald ripening. The shapes of their particle size distributions can yield the rate law for this process. One consequence of Ostwald ripening is that a record of the recrystallization process is preserved in the various particle sizes. Therefore, one can determine the detailed geologic history of clays and other recrystallized minerals by separating, from a single sample, the various particle sizes for independent chemical, structural, and isotopic analyses.

  6. The influence of ripening temperature on ‘Hass’ fruit quality

    USDA-ARS?s Scientific Manuscript database

    Previous research demonstrated the importance of temperature management during avocado (Persea americana Mill) fruit ripening; however this work was focused on maintaining the fruit at elevated temperatures continuously during the ripening process. We examined the influence of short duration high t...

  7. Changes in Gene Expression during Tomato Fruit Ripening 1

    PubMed Central

    Biggs, M. Scott; Harriman, Robert W.; Handa, Avtar K.

    1986-01-01

    Total proteins from pericarp tissue of different chronological ages from normally ripening tomato (Lycopersicon esculentum Mill. cv Rutgers) fruits and from fruits of the isogenic ripening-impaired mutants rin, nor, and Nr were extracted and separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Analysis of the stained bands revealed increases in 5 polypeptides (94, 44, 34, 20, and 12 kilodaltons), decreases in 12 polypeptides (106, 98, 88, 76, 64, 52, 48, 45, 36, 28, 25, and 15 kilodaltons), and fluctuations in 5 polypeptides (85, 60, 26, 21, and 16 kilodaltons) as normal ripening proceeded. Several polypeptides present in ripening normal pericarp exhibited very low or undetectable levels in developing mutant pericarp. Total RNAs extracted from various stages of Rutgers pericarp and from 60 to 65 days old rin, nor, and Nr pericarp were fractionated into poly(A)+ and poly(A)− RNAs. Peak levels of total RNA, poly(A)+ RNA, and poly(A)+ RNA as percent of total RNA occurred between the mature green to breaker stages of normal pericarp. In vitro translation of poly(A)+ RNAs from normal pericarp in rabbit reticulocyte lysates revealed increases in mRNAs for 9 polypeptides (116, 89, 70, 42, 38, 33, 31, 29, and 26 kilodaltons), decreases in mRNAs for 2 polypeptides (41 and 35 kilodaltons), and fluctuations in mRNAs for 5 polypeptides (156, 53, 39, 30, and 14 kilodaltons) during normal ripening. Analysis of two-dimensional separation of in vitro translated polypeptides from poly(A)+ RNAs isolated from different developmental stages revealed even more extensive changes in mRNA populations during ripening. In addition, a polygalacturonase precursor (54 kilodaltons) was immunoprecipitated from breaker, turning, red ripe, and 65 days old Nr in vitro translation products. Images Fig. 1 Fig. 3 Fig. 5 Fig. 6 Fig. 7 PMID:16664828

  8. Some Properties of Fresh and Ripened Traditional Akcakatik Cheese

    PubMed Central

    2018-01-01

    Akcakatik cheese (yogurt cheese) is produced by drying strained yogurt with or without adding cloves or black cumin. The main objective of this study was to detect the properties of both fresh and ripened Akcakatik cheeses and to compare them. For this purpose the biogenic amine content, volatile flavor compounds, protein degradation level, chemical properties and some microbiological properties of 15 Akcakatik cheese samples were investigated. Titratable acidity, total dry matter, NaCl, total nitrogen, water soluble nitrogen, ripened index, histamine, diacetyl and acetaldehyde levels were found to be higher in ripened cheese samples than in fresh cheese samples. On the other hand, the clove and black cumin ratios were found to be higher in the fresh cheese samples. Sodium dodecyl sulphate polyacrylamide gel electropherograms of cheese samples showed that protein degradation was higher in ripened cheese samples than in fresh samples, as expected. The dominant Lactic acid bacteria (LAB) flora of Akcakatik cheese samples were found to be Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. PMID:29725229

  9. Identification and antioxidant properties of polyphenols in lotus seed epicarp at different ripening stages.

    PubMed

    Liu, Ying; Ma, Shuang-shuang; Ibrahim, S A; Li, Er-hu; Yang, Hong; Huang, Wen

    2015-10-15

    In this study, polyphenols from lotus seed epicarp (PLSE) at three different ripening stages were purified by column chromatography and identified by RP-HPLC and HPLC-ESI-MS(2). The antioxidant activities of PLSE were also investigated. We found that the contents of PLSE at the green ripening stage, half ripening stage and full ripening stage are 13.08%, 10.95% and 6.73% respectively. The levels of catechin, epicatechin, hyperoside, and isoquercitrin in PLSE at the three different ripening stages were different. Moreover, the amounts of catechin and epicatechin decreased, while the contents of hyperoside and isoquercitrin increased as the seed ripened. We found that PLSE at three different ripening stages had good scavenging abilities on DPPH and ABTS(+) radicals. However, the scavenging ability decreased with maturation. Our results may be valuable with regard to the utilization of lotus seed epicarp as a functional food material. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Natural Variation in Banana Varieties Highlights the Role of Melatonin in Postharvest Ripening and Quality.

    PubMed

    Hu, Wei; Yang, Hai; Tie, Weiwei; Yan, Yan; Ding, Zehong; Liu, Yang; Wu, Chunlai; Wang, Jiashui; Reiter, Russel J; Tan, Dun-Xian; Shi, Haitao; Xu, Biyu; Jin, Zhiqiang

    2017-11-22

    This study aimed to investigate the role of melatonin in postharvest ripening and quality in various banana varieties with contrasting ripening periods. During the postharvest life, endogenous melatonin showed similar performance with ethylene in connection to ripening. In comparison to ethylene, melatonin was more correlated with postharvest banana ripening. Exogenous application of melatonin resulted in a delay of postharvest banana ripening. Moreover, this effect is concentration-dependent, with 200 and 500 μM treatments more effective than the 50 μM treatment. Exogenous melatonin also led to elevated endogenous melatonin content, reduced ethylene production through regulation of the expression of MaACO1 and MaACS1, and delayed sharp changes of quality indices. Taken together, this study highlights that melatonin is an indicator for banana fruit ripening in various varieties, and the repression of ethylene biosynthesis and postharvest ripening by melatonin can be used for biological control of postharvest fruit ripening and quality.

  11. Transcriptional analysis of late ripening stages of grapevine berry

    PubMed Central

    2011-01-01

    Background The composition of grapevine berry at harvest is a major determinant of wine quality. Optimal oenological maturity of berries is characterized by a high sugar/acidity ratio, high anthocyanin content in the skin, and low astringency. However, harvest time is still mostly determined empirically, based on crude biochemical composition and berry tasting. In this context, it is interesting to identify genes that are expressed/repressed specifically at the late stages of ripening and which may be used as indicators of maturity. Results Whole bunches and berries sorted by density were collected in vineyard on Chardonnay (white cultivar) grapevines for two consecutive years at three stages of ripening (7-days before harvest (TH-7), harvest (TH), and 10-days after harvest (TH+10)). Microvinification and sensory analysis indicate that the quality of the wines made from the whole bunches collected at TH-7, TH and TH+10 differed, TH providing the highest quality wines. In parallel, gene expression was studied with Qiagen/Operon microarrays using two types of samples, i.e. whole bunches and berries sorted by density. Only 12 genes were consistently up- or down-regulated in whole bunches and density sorted berries for the two years studied in Chardonnay. 52 genes were differentially expressed between the TH-7 and TH samples. In order to determine whether these genes followed a similar pattern of expression during the late stages of berry ripening in a red cultivar, nine genes were selected for RT-PCR analysis with Cabernet Sauvignon grown under two different temperature regimes affecting the precocity of ripening. The expression profiles and their relationship to ripening were confirmed in Cabernet Sauvignon for seven genes, encoding a carotenoid cleavage dioxygenase, a galactinol synthase, a late embryogenesis abundant protein, a dirigent-like protein, a histidine kinase receptor, a valencene synthase and a putative S-adenosyl-L-methionine:salicylic acid carboxyl

  12. Decrease in fruit moisture content heralds and might launch the onset of ripening processes.

    PubMed

    Frenkel, Chaim; Hartman, Thomas G

    2012-10-01

    It is known that fruit ripening is a genetically programmed event but it is not entirely clear what metabolic cue(s) stimulate the onset of ripening, ethylene action notwithstanding. Here, we examined the conjecture that fruit ripening might be evoked by an autonomously induced decrease in tissue water status. We found decline in water content occurring at the onset of ripening in climacteric and nonclimacteric fruit, suggesting that this phenomenon might be universal. This decline in water content persisted throughout the ripening process in some fruit, whereas in others it reversed during the progression of the ripening process. Applied ethylene also induced a decrease in water content in potato (Solanum tuberosum) tubers. In ethylene-mutant tomato (Solanum lycopersicum) fruit (antisense to1-aminocyclopropane carboxylate synthase), cold-induced decline in water content stimulated onset of ripening processes apparently independently of ethylene action, suggesting cause-and-effect relationship between decreasing water content and onset of ripening. The decline in tissue water content, occurring naturally or induced by ethylene, was strongly correlated with a decrease in hydration (swelling) efficacy of cell wall preparations suggesting that hydration dynamics of cell walls might account for changes in tissue moisture content. Extent of cell wall swelling was, in turn, related to the degree of oxidative cross-linking of wall-bound phenolic acids, suggesting that oxidant-induced wall restructuring might mediate cell wall and, thus, fruit tissue hydration status. We propose that oxidant-induced cell wall remodeling and consequent wall dehydration might evoke stress signaling for the onset of ripening processes. This study suggests that decline in fruit water content is an early event in fruit ripening. This information may be used to gauge fruit maturity for appropriate harvest date and for processing. Control of fruit hydration state might be used to regulate the

  13. Effects of Pu-erh ripened tea on hyperuricemic mice studied by serum metabolomics.

    PubMed

    Zhao, Ran; Chen, Dong; Wu, Hualing

    2017-11-15

    To evaluate effects of Pu-erh ripened tea in hyperuricemic mice, a mouse hyperuricemia model was developed by oral administration of potassium oxonate for 7 d. Serum metabolomics, based on gas chromatography-mass spectrometry, was used to generate metabolic profiles from normal control, hyperuricemic and allopurinol-treated hyperuricemic mice, as well as hyperuricemic mice given Pu-erh ripened tea at three doses. Pu-erh ripened tea significantly lowered serum uric acid levels. Twelve potential biomarkers associated with hyperuricemia were identified. Pu-erh ripened tea and allopurinol differed in their metabolic effects in the hyperuricemic mice. Levels of glutamic acid, indolelactate, L-allothreonine, nicotinoylglycine, isoleucine, l-cysteine and glycocyamine, all involved in amino acid metabolism, were significantly changed in hyperuricemic mice treated Pu-erh ripened tea. Thus, modulating amino acid metabolism might be the primary mechanism of anti-hyperuricemia by Pu-erh ripened tea. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Olive oil quality and ripening in super-high-density Arbequina orchard.

    PubMed

    Benito, Marta; Lasa, José Manuel; Gracia, Pilar; Oria, Rosa; Abenoza, María; Varona, Luis; Sánchez-Gimeno, Ana Cristina

    2013-07-01

    The aim of this work was to evaluate the evolution of the quality of extra virgin olive oil obtained from a super-high-density Arbequina orchard, under a drip irrigation system, throughout the ripening process. For this objective, physicochemical, nutritional and sensory parameters were studied. In addition, the oxidative stability, pigment content and colour evolution of olive oil were analysed during the ripening process. Free acidity increased slightly throughout the ripening process, while peroxide value and extinction coefficient decreased. Total phenol content and oxidative stability showed a similar trend, increasing at the beginning of ripening up to a maximum and thereafter decreasing. α-Tocopherol and pigment contents decreased with ripening, leading to changes in colour coordinates. Sensory parameters were correlated with total phenol content, following a similar trend throughout the maturation process. By sampling and monitoring the ripeness index weekly, it would be possible to determine an optimal harvesting time for olives according to the industrial yield and the physicochemical, nutritional and sensory properties of the olive oil. © 2012 Society of Chemical Industry.

  15. Complex interactive effects of ripening degree, malaxation duration and temperature on Oblica cv. virgin olive oil phenols, volatiles and sensory quality.

    PubMed

    Lukić, Igor; Žanetić, Mirella; Jukić Špika, Maja; Lukić, Marina; Koprivnjak, Olivera; Brkić Bubola, Karolina

    2017-10-01

    The interactive effects of ripening degree, malaxation duration and temperature on Oblica cv. (Olea europaea L.) virgin olive oil phenols, volatiles, and sensory quality were investigated. Olives were picked at three ripening degrees with International Olive Council indices of 0.68, 2.48 and 4.10, and processed by malaxation at 22 and 30°C, and at both temperatures for 30 and 60min. Ripening exhibited the strongest effect, and malaxation duration the weakest. Phenols were generally found to decrease during ripening; however 3,4-DHPEA-EDA and p-HPEA-EDA increased. Similar behaviour was observed for (E)-2-hexenal. Higher malaxation temperature induced an increase in particular important phenols and C6 alcohols, while C6 aldehydes mostly decreased. Interactions between the factors were established, mostly between ripening degree and malaxation temperature: the effect of the latter was most pronounced for ripe olives, especially for 3,4-DHPEA-EDA, p-HPEA-EDA and C6 volatiles. Sensory attributes were generally in agreement with the chemical composition. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Sphingolipid Distribution, Content and Gene Expression during Olive-Fruit Development and Ripening

    PubMed Central

    Inês, Carla; Parra-Lobato, Maria C.; Paredes, Miguel A.; Labrador, Juana; Gallardo, Mercedes; Saucedo-García, Mariana; Gavilanes-Ruiz, Marina; Gomez-Jimenez, Maria C.

    2018-01-01

    Plant sphingolipids are involved in the building of the matrix of cell membranes and in signaling pathways of physiological processes and environmental responses. However, information regarding their role in fruit development and ripening, a plant-specific process, is unknown. The present study seeks to determine whether and, if so, how sphingolipids are involved in fleshy-fruit development and ripening in an oil-crop species such as olive (Olea europaea L. cv. Picual). Here, in the plasma-membranes of live protoplasts, we used fluorescence to examine various specific lipophilic stains in sphingolipid-enriched regions and investigated the composition of the sphingolipid long-chain bases (LCBs) as well as the expression patterns of sphingolipid-related genes, OeSPT, OeSPHK, OeACER, and OeGlcCerase, during olive-fruit development and ripening. The results demonstrate increased sphingolipid content and vesicle trafficking in olive-fruit protoplasts at the onset of ripening. Moreover, the concentration of LCB [t18:1(8Z), t18:1 (8E), t18:0, d18:2 (4E/8Z), d18:2 (4E/8E), d18:1(4E), and 1,4-anhydro-t18:1(8E)] increases during fruit development to reach a maximum at the onset of ripening, although these molecular species decreased during fruit ripening. On the other hand, OeSPT, OeSPHK, and OeGlcCerase were expressed differentially during fruit development and ripening, whereas OeACER gene expression was detected only at the fully ripe stage. The results provide novel data about sphingolipid distribution, content, and biosynthesis/turnover gene transcripts during fleshy-fruit ripening, indicating that all are highly regulated in a developmental manner. PMID:29434611

  17. Changes in the sensory perception of 'Hass' avocado during fruit maturation and ripening

    USDA-ARS?s Scientific Manuscript database

    Avocado flavor improves dramatically during maturation and also changes as a result of ripening, but prior research had focused almost entirely on alterations in oil content as a means of explaining the differences in flavor. Other factors beyond just oil content are likely involved that act to alt...

  18. Growth of Aerobic Ripening Bacteria at the Cheese Surface Is Limited by the Availability of Iron

    PubMed Central

    Back, Alexandre; Irlinger, Françoise

    2012-01-01

    The microflora on the surface of smear-ripened cheeses is composed of various species of bacteria and yeasts that contribute to the production of the desired organoleptic properties. The objective of the present study was to show that iron availability is a limiting factor in the growth of typical aerobic ripening bacteria in cheese. For that purpose, we investigated the effect of iron or siderophore addition in model cheeses that were coinoculated with a yeast and a ripening bacterium. Both iron and the siderophore desferrioxamine B stimulated the growth of ripening bacteria belonging to the genera Arthrobacter, Corynebacterium, and Brevibacterium. The extent of stimulation was strain dependent, and generally, the effect of desferrioxamine B was greater than that of iron. Measurements of the expression of genes related to the metabolism of iron by Arthrobacter arilaitensis Re117 by real-time reverse transcription-PCR showed that these genes were transcribed during growth in cheese. The addition of desferrioxamine B increased the expression of two genes encoding iron-siderophore ABC transport binding proteins. The addition of iron decreased the expression of siderophore biosynthesis genes and of part of the genes encoding iron-siderophore ABC transport components. It was concluded that iron availability is a limiting factor in the growth of typical cheese surface bacteria. The selection of strains with efficient iron acquisition systems may be useful for the development of defined-strain surface cultures. Furthermore, the importance of iron metabolism in the microbial ecology of cheeses should be investigated since it may result in positive or negative microbial interactions. PMID:22367081

  19. Overview of a Surface-Ripened Cheese Community Functioning by Meta-Omics Analyses

    PubMed Central

    Teissandier, Aurélie; Onésime, Djamila; Loux, Valentin; Monnet, Christophe; Irlinger, Françoise; Landaud, Sophie; Leclercq-Perlat, Marie-Noëlle; Bento, Pascal; Fraud, Sébastien; Gibrat, Jean-François; Aubert, Julie; Fer, Frédéric; Guédon, Eric; Pons, Nicolas; Kennedy, Sean; Beckerich, Jean-Marie; Swennen, Dominique; Bonnarme, Pascal

    2015-01-01

    Cheese ripening is a complex biochemical process driven by microbial communities composed of both eukaryotes and prokaryotes. Surface-ripened cheeses are widely consumed all over the world and are appreciated for their characteristic flavor. Microbial community composition has been studied for a long time on surface-ripened cheeses, but only limited knowledge has been acquired about its in situ metabolic activities. We applied metagenomic, metatranscriptomic and biochemical analyses to an experimental surface-ripened cheese composed of nine microbial species during four weeks of ripening. By combining all of the data, we were able to obtain an overview of the cheese maturation process and to better understand the metabolic activities of the different community members and their possible interactions. Furthermore, differential expression analysis was used to select a set of biomarker genes, providing a valuable tool that can be used to monitor the cheese-making process. PMID:25867897

  20. Determining the minimum ripening time of artisanal Minas cheese, a traditional Brazilian cheese

    PubMed Central

    Martins, José M.; Galinari, Éder; Pimentel-Filho, Natan J.; Ribeiro, José I.; Furtado, Mauro M.; Ferreira, Célia L.L.F.

    2015-01-01

    Physical, physicochemical, and microbiological changes were monitored in 256 samples of artisanal Minas cheese from eight producers from Serro region (Minas Gerais, Brazil) for 64 days of ripening to determine the minimum ripening time for the cheese to reach the safe microbiological limits established by Brazilian legislation. The cheeses were produced between dry season (April–September) and rainy season (October–March); 128 cheeses were ripened at room temperature (25 ± 4 °C), and 128 were ripened under refrigeration (8 ± 1 °C), as a control. No Listeria monocytogenes was found, but one cheese under refrigeration had Salmonella at first 15 days of ripening. However, after 22 days, the pathogen was not detected. Seventeen days was the minimum ripening time at room temperature to reduce at safe limits of total coliforms > 1000 cfu.g −1 ), Escherichia coli and Staphylococcus aureus (> 100 cfu.g −1 ) in both periods of manufacture. Otherwise under refrigeration, as expected, the minimum ripening time was longer, 33 days in the dry season and 63 days in the rainy season. To sum up, we suggest that the ripening of artisanal Minas cheese be done at room temperature, since this condition shortens the time needed to reach the microbiological quality that falls within the safety parameters required by Brazilian law, and at the same time maintain the appearance and flavor characteristics of this traditional cheese. PMID:26221111

  1. Acquired high titre factor VIII inhibitor with underlying polyarteritis nodosa.

    PubMed

    Snowden, J A; Hutchings, M; Spearing, R; Patton, W N

    1997-05-01

    We here present the case of a 70-year-old woman referred to our unit for investigation of bleeding. Investigations confirmed a high titre acquired Factor VIII inhibitor. In association there was relapse of systemic illness associated with anti-neutrophil cytoplasmic antibodies (atypical pattern) for which she had been treated five years previously. Immunosuppression was attempted, but it failed to have an impact both on the inhibitor titre and on the underlying disorder. The patient died from multi-organ failure and massive chest hemorrhage. Post-mortem showed necrotizing vasculitis of medium sized vessels at several sites, including the kidney, consistent with a diagnosis of polyarteritis nodosa. Although it is well recognised that Factor VIII inhibitors are found in conjunction with autoimmune disorders, this case is significant in that it is the first associated with histologically proven polyarteritis nodosa type vasculitis. The case illustrates the difficulties in the investigation and management of patients with acquired high titre Factor VIII inhibitors.

  2. Transcript and metabolite analysis in Trincadeira cultivar reveals novel information regarding the dynamics of grape ripening

    PubMed Central

    2011-01-01

    metabolism. These results were integrated with transcriptional profiling obtained using genome array to provide new information regarding the network of events leading to grape ripening. Conclusions Altogether the data obtained provides the most extensive survey obtained so far for gene expression and metabolites accumulated during grape ripening. Moreover, it highlighted information obtained in a poorly known variety exhibiting particular characteristics that may be cultivar specific or dependent upon climatic conditions. Several genes were identified that had not been previously reported in the context of grape ripening namely genes involved in carbohydrate and amino acid metabolisms as well as in growth regulators; metabolism, epigenetic factors and signaling pathways. Some of these genes were annotated as receptors, transcription factors, and kinases and constitute good candidates for functional analysis in order to establish a model for ripening control of a non-climacteric fruit. PMID:22047180

  3. Transcript and metabolite analysis in Trincadeira cultivar reveals novel information regarding the dynamics of grape ripening.

    PubMed

    Fortes, Ana M; Agudelo-Romero, Patricia; Silva, Marta S; Ali, Kashif; Sousa, Lisete; Maltese, Federica; Choi, Young H; Grimplet, Jerome; Martinez-Zapater, José M; Verpoorte, Robert; Pais, Maria S

    2011-11-02

    results were integrated with transcriptional profiling obtained using genome array to provide new information regarding the network of events leading to grape ripening. Altogether the data obtained provides the most extensive survey obtained so far for gene expression and metabolites accumulated during grape ripening. Moreover, it highlighted information obtained in a poorly known variety exhibiting particular characteristics that may be cultivar specific or dependent upon climatic conditions. Several genes were identified that had not been previously reported in the context of grape ripening namely genes involved in carbohydrate and amino acid metabolisms as well as in growth regulators; metabolism, epigenetic factors and signaling pathways. Some of these genes were annotated as receptors, transcription factors, and kinases and constitute good candidates for functional analysis in order to establish a model for ripening control of a non-climacteric fruit.

  4. Circadian oscillatory transcriptional programs in grapevine ripening fruits

    PubMed Central

    2014-01-01

    Background Temperature and solar radiation influence Vitis vinifera L. berry ripening. Both environmental conditions fluctuate cyclically on a daily period basis and the strength of this fluctuation affects grape ripening too. Additionally, a molecular circadian clock regulates daily cyclic expression in a large proportion of the plant transcriptome modulating multiple developmental processes in diverse plant organs and developmental phases. Circadian cycling of fruit transcriptomes has not been characterized in detail despite their putative relevance in the final composition of the fruit. Thus, in this study, gene expression throughout 24 h periods in pre-ripe berries of Tempranillo and Verdejo grapevine cultivars was followed to determine whether different ripening transcriptional programs are activated during certain times of day in different grape tissues and genotypes. Results Microarray analyses identified oscillatory transcriptional profiles following circadian variations in the photocycle and the thermocycle. A higher number of expression oscillating transcripts were detected in samples carrying exocarp tissue including biotic stress-responsive transcripts activated around dawn. Thermotolerance-like responses and regulation of circadian clock-related genes were observed in all studied samples. Indeed, homologs of core clock genes were identified in the grapevine genome and, among them, VvREVEILLE1 (VvRVE1), showed a consistent circadian expression rhythm in every grape berry tissue analysed. Light signalling components and terpenoid biosynthetic transcripts were specifically induced during the daytime in Verdejo, a cultivar bearing white-skinned and aromatic berries, whereas transcripts involved in phenylpropanoid biosynthesis were more prominently regulated in Tempranillo, a cultivar bearing black-skinned berries. Conclusions The transcriptome of ripening fruits varies in response to daily environmental changes, which might partially be under the control

  5. Non-climacteric ripening and sorbitol homeostasis in plum fruits.

    PubMed

    Kim, Ho-Youn; Farcuh, Macarena; Cohen, Yuval; Crisosto, Carlos; Sadka, Avi; Blumwald, Eduardo

    2015-02-01

    During ripening fruits undergo several physiological and biochemical modifications that influence quality-related properties, such as texture, color, aroma and taste. We studied the differences in ethylene and sugar metabolism between two genetically related Japanese plum cultivars with contrasting ripening behaviors. 'Santa Rosa' (SR) behaved as a typical climacteric fruit, while the bud sport mutant 'Sweet Miriam' (SM) displayed a non-climacteric ripening pattern. SM fruit displayed a delayed ripening that lasted 120 days longer than that of the climacteric fruit. At the full-ripe stage, both cultivars reached similar final size and weight but the non-climacteric fruits were firmer than the climacteric fruits. Fully ripe non-climacteric plum fruits, showed an accumulation of sorbitol that was 2.5 times higher than that of climacteric fruits, and the increase in sorbitol were also paralleled to an increase in sucrose catabolism. These changes were highly correlated with decreased activity and expression of NAD(+)-dependent sorbitol dehydrogenase and sorbitol oxidase and increased sorbitol-6-phosphate dehydrogenase activity, suggesting an enhanced sorbitol synthesis in non-climacteric fruits. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  6. Polyphenol Compound as a Transcription Factor Inhibitor.

    PubMed

    Park, Seyeon

    2015-10-30

    A target-based approach has been used to develop novel drugs in many therapeutic fields. In the final stage of intracellular signaling, transcription factor-DNA interactions are central to most biological processes and therefore represent a large and important class of targets for human therapeutics. Thus, we focused on the idea that the disruption of protein dimers and cognate DNA complexes could impair the transcriptional activation and cell transformation regulated by these proteins. Historically, natural products have been regarded as providing the primary leading compounds capable of modulating protein-protein or protein-DNA interactions. Although their mechanism of action is not fully defined, polyphenols including flavonoids were found to act mostly as site-directed small molecule inhibitors on signaling. There are many reports in the literature of screening initiatives suggesting improved drugs that can modulate the transcription factor interactions responsible for disease. In this review, we focus on polyphenol compound inhibitors against dimeric forms of transcription factor components of intracellular signaling pathways (for instance, c-jun/c-fos (Activator Protein-1; AP-1), c-myc/max, Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and β-catenin/T cell factor (Tcf)).

  7. Comparative Transcriptome Analysis of Climacteric Fruit of Chinese Pear (Pyrus ussuriensis) Reveals New Insights into Fruit Ripening

    PubMed Central

    Tan, Dongmei; Jiang, Zhongyu; Wei, Yun; Li, Juncai; Wang, Aide

    2014-01-01

    The fruit of Pyrus ussuriensis is typically climacteric. During ripening, the fruits produce a large amount of ethylene, and their firmness drops rapidly. Although the molecular basis of climacteric fruit ripening has been studied in depth, some aspects remain unclear. Here, we compared the transcriptomes of pre- and post-climacteric fruits of Chinese pear (P. ussuriensis c.v. Nanguo) using RNA-seq. In total, 3,279 unigenes were differentially expressed between the pre- and post-climacteric fruits. Differentially expressed genes (DEGs) were subjected to Gene Ontology analysis, and 31 categories were significantly enriched in the groups ‘biological process’, ‘molecular function’ and ‘cellular component’. The DEGs included genes related to plant hormones, such as ethylene, ABA, auxin, GA and brassinosteroid, and transcription factors, such as MADS, NAC, WRKY and HSF. Moreover, genes encoding enzymes related to DNA methylation, cytoskeletal proteins and heat shock proteins (HSPs) showed differential expression between the pre- and post-climacteric fruits. Select DEGs were subjected to further analysis using quantitative RT-PCR (qRT-PCR), and the results were consistent with those of RNA-seq. Our data suggest that in addition to ethylene, other hormones play important roles in regulating fruit ripening and may interact with ethylene signaling during this process. DNA methylation-related methyltransferase and cytoskeletal protein genes are also involved in fruit ripening. Our results provide useful information for future research on pear fruit ripening. PMID:25215597

  8. How Do Fruits Ripen?

    ERIC Educational Resources Information Center

    Sargent, Steven A.

    2005-01-01

    A fruit is alive, and for it to ripen normally, many biochemical reactions must occur in a proper order. After pollination, proper nutrition, growing conditions, and certain plant hormones cause the fruit to develop and grow to proper size. During this time, fruits store energy in the form of starch and sugars, called photosynthates because they…

  9. Fruit development and ripening.

    PubMed

    Seymour, Graham B; Østergaard, Lars; Chapman, Natalie H; Knapp, Sandra; Martin, Cathie

    2013-01-01

    Fruiting structures in the angiosperms range from completely dry to highly fleshy organs and provide many of our major crop products, including grains. In the model plant Arabidopsis, which has dry fruits, a high-level regulatory network of transcription factors controlling fruit development has been revealed. Studies on rare nonripening mutations in tomato, a model for fleshy fruits, have provided new insights into the networks responsible for the control of ripening. It is apparent that there are strong similarities between dry and fleshy fruits in the molecular circuits governing development and maturation. Translation of information from tomato to other fleshy-fruited species indicates that regulatory networks are conserved across a wide spectrum of angiosperm fruit morphologies. Fruits are an essential part of the human diet, and recent developments in the sequencing of angiosperm genomes have provided the foundation for a step change in crop improvement through the understanding and harnessing of genome-wide genetic and epigenetic variation.

  10. Comparative Transcriptome Analyses between a Spontaneous Late-Ripening Sweet Orange Mutant and Its Wild Type Suggest the Functions of ABA, Sucrose and JA during Citrus Fruit Ripening

    PubMed Central

    Zhang, Ya-Jian; Wang, Xing-Jian; Wu, Ju-Xun; Chen, Shan-Yan; Chen, Hong; Chai, Li-Jun; Yi, Hua-Lin

    2014-01-01

    A spontaneous late-ripening mutant of ‘Jincheng’ (C. sinensis L. Osbeck) sweet orange exhibited a delay of fruit pigmentation and harvesting. In this work, we studied the processes of orange fruit ripening through the comparative analysis between the Jincheng mutant and its wild type. This study revealed that the fruit quality began to differ on 166th days after anthesis. At this stage, fruits were subjected to transcriptome analysis by RNA sequencing. 13,412 differentially expressed unigenes (DEGs) were found. Of these unigenes, 75.8% were down-regulated in the wild type, suggesting that the transcription level of wild type was lower than that of the mutant during this stage. These DEGs were mainly clustered into five pathways: metabolic pathways, plant-pathogen interaction, spliceosome, biosynthesis of plant hormones and biosynthesis of phenylpropanoids. Therefore, the expression profiles of the genes that are involved in abscisic acid, sucrose, and jasmonic acid metabolism and signal transduction pathways were analyzed during the six fruit ripening stages. The results revealed the regulation mechanism of sweet orange fruit ripening metabolism in the following four aspects: First, the more mature orange fruits were, the lower the transcription levels were. Second, the expression level of PME boosted with the maturity of the citrus fruit. Therefore, the expression level of PME might represent the degree of the orange fruit ripeness. Third, the interaction of PP2C, PYR/PYL, and SnRK2 was peculiar to the orange fruit ripening process. Fourth, abscisic acid, sucrose, and jasmonic acid all took part in orange fruit ripening process and might interact with each other. These findings provide an insight into the intricate process of sweet orange fruit ripening. PMID:25551568

  11. The role of alternative oxidase in tomato fruit ripening and its regulatory interaction with ethylene

    PubMed Central

    Lin, Hong-Hui

    2012-01-01

    Although the alternative oxidase (AOX) has been proposed to play a role in fruit development, the function of AOX in fruit ripening is unclear. To gain further insight into the role of AOX in tomato fruit ripening, transgenic tomato plants 35S-AOX1a and 35S-AOX-RNAi were generated. Tomato plants with reduced LeAOX levels exhibited retarded ripening; reduced carotenoids, respiration, and ethylene production; and the down-regulation of ripening-associated genes. Moreover, no apparent respiratory climacteric occurred in the AOX-reduced tomato fruit, indicating that AOX might play an important role in climacteric respiration. In contrast, the fruit that overexpressed LeAOX1a accumulated more lycopene, though they displayed a similar pattern of ripening to wild-type fruit. Ethylene application promoted fruit ripening and anticipated ethylene production and respiration, including the alternative pathway respiration. Interestingly, the transgenic plants with reduced LeAOX levels failed to ripen after 1-methylcyclopropene (1-MCP) treatment, while such inhibition was notably less effective in 35S-AOX1a fruit. These findings indicate that AOX is involved in respiratory climacteric and ethylene-mediated fruit ripening of tomato. PMID:22915749

  12. A close insight to factor VIII inhibitor in the congenital hemophilia A.

    PubMed

    Tabriznia-Tabrizi, Shamsoreza; Gholampour, Marzie; Mansouritorghabeh, Hassan

    2016-09-01

    Hemophilia A (HA) has an X-linked pattern of inheritance and is the most common of the hemorrhagic disorders. HA is caused by a decreased or deficiency of the functional clotting factor VIII (FVIII) and effects 1 in 5000-10,000 male births. The common treatment for hemophilia is replacement therapy by plasma-derived or recombinant FVIII. Approximately 20-30% of people with a severe type of HA develop an inhibitor and this phenomenon is the main challenge in the management of these patients. Genetic factors and environmental determinants contribute to inhibitor development. Here, the roles of various genetic and environmental factors such as the type of FVIII concentrate used, the number of exposure days, and peak treatment time will be discussed in detail. It seems this information is helpful for hematologists. A literature review was done in January 2016 on PubMed and Scopus using the following keywords:' h(a)emophilia A & factor VIII inhibitor', 'h(a)emophilia A & factor VIII alloantibody', 'h(a)emophilia A & inhibitor'. There was no time limitation; however, there was an English language limitation placed on the articles selected. Expert commentary: Influential genetic and environmental factors in developing inhibitors have been discussed. Most of the risk factors are related to previously untreated patients with hemophili.

  13. Physiological characteristics and related gene expression of after-ripening on seed dormancy release in rice.

    PubMed

    Du, W; Cheng, J; Cheng, Y; Wang, L; He, Y; Wang, Z; Zhang, H

    2015-11-01

    After-ripening is a common method used for dormancy release in rice. In this study, the rice variety Jiucaiqing (Oryza sativa L. subsp. japonica) was used to determine dormancy release following different after-ripening times (1, 2 and 3 months). Germination speed, germination percentage and seedling emergence increased with after-ripening; more than 95% germination and 85% seedling emergence were observed following 1 month of after-ripening within 10 days of imbibition, compared with <45% germination and 20% seedling emergence in freshly harvested seed. Hence, 3 months of after-ripening could be considered a suitable treatment period for rice dormancy release. Dormancy release by after-ripening is mainly correlated with a rapid decline in ABA content and increase in IAA content during imbibition. Subsequently, GA(1)/ABA, GA(7)/ABA, GA(12)/ABA, GA(20)/ABA and IAA/ABA ratios significantly increased, while GA(3)/ABA, GA(4)/ABA and GAs/IAA ratio significantly decreased in imbibed seeds following 3 months of after-ripening, thereby altering α-amylase activity during seed germination. Peak α-amylase activity occurred at an earlier germination stage in after-ripened seeds than in freshly harvested seeds. Expression of ABA, GA and IAA metabolism genes and dormancy-related genes was regulated by after-ripening time upon imbibition. Expression of OsCYP707A5, OsGA2ox1, OsGA2ox2, OsGA2ox3, OsILR1, OsGH3-2, qLTG3-1 and OsVP1 increased, while expression of Sdr4 decreased in imbibed seeds following 3 months of after-ripening. Dormancy release through after-ripening might be involved in weakening tissues covering the embryo via qLTG3-1 and decreased ABA signalling and sensitivity via Sdr4 and OsVP1. © 2015 German Botanical Society and The Royal Botanical Society of the Netherlands.

  14. Edible coatings influence fruit ripening, quality, and aroma biosynthesis in mango fruit.

    PubMed

    Dang, Khuyen T H; Singh, Zora; Swinny, Ewald E

    2008-02-27

    The effects of different edible coatings on mango fruit ripening and ripe fruit quality parameters including color, firmness, soluble solids concentrations, total acidity, ascorbic acid, total carotenoids, fatty acids, and aroma volatiles were investigated. Hard mature green mango (Mangifera indica L. cv. Kensigton Pride) fruits were coated with aqueous mango carnauba (1:1 v/v), Semperfresh (0.6%), Aloe vera gel (1:1, v/v), or A. vera gel (100%). Untreated fruit served as the control. Following the coating, fruits were allowed to dry at room temperature and packed in soft-board trays to ripen at 21+/-1 degrees C and 55.2+/-11.1% relative humidity until the eating soft stage. Mango carnauba was effective in retarding fruit ripening, retaining fruit firmness, and improving fruit quality attributes including levels of fatty acids and aroma volatiles. Semperfresh and A. vera gel (1:1 or 100%) slightly delayed fruit ripening but reduced fruit aroma volatile development. A. vera gel coating did not exceed the commercial mango carnauba and Semperfresh in retarding fruit ripening and improving aroma volatile biosynthesis.

  15. Anthrax lethal factor inhibitors as potential countermeasure of the infection.

    PubMed

    Kumar, B V S Suneel; Malik, Siddharth; Grandhi, Pradeep; Dayam, Raveendra; Sarma, J A R P

    2014-01-01

    Anthrax Lethal Factor (LF) is a zinc-dependent metalloprotease, one of the virulence factor of anthrax infection. Three forms of the anthrax infection have been identified: cutaneous (through skin), gastrointestinal (through alimentary tract), and pulmonary (by inhalation of spores). Anthrax toxin is composed of protective antigen (PA), lethal factor (LF), and edema factor (EF). Protective antigen mediates the entry of Lethal Factor/Edema Factor into the cytosol of host cells. Lethal factor (LF) inactivates mitogen-activated protein kinase kinase inducing cell death, and EF is an adenylyl cyclase impairing host defenses. In the past few years, extensive studies are undertaken to design inhibitors targeting LF. The current review focuses on the small molecule inhibitors targeting LF activity and its structure activity relationships (SAR).

  16. Detection and Viability of Lactococcus lactis throughout Cheese Ripening

    PubMed Central

    Cocolin, Luca

    2014-01-01

    Recent evidences highlighted the presence of Lactococcus lactis during late cheese ripening. For this reason, the role of this microorganism, well known as dairy starter, should be reconsidered throughout cheese manufacturing and ripening. Thus, the main objective of this study was to develop a RT-qPCR protocol for the detection, quantification and determination of the viability of L. lactis in ripened cheese samples by direct analysis of microbial nucleic acids. Standard curves were constructed for the specific quantification of L. lactis in cheese matrices and good results in terms of selectivity, correlation coefficient and efficiency were obtained. Thirty-three ripened cheeses were analyzed and, on the basis of RNA analysis, twelve samples showed 106 to 108 CFU of L. lactis per gram of product, thirteen from 103 to 105 CFU/g, and in eight cheeses, L. lactis was not detected. Traditional plating on M17 medium led to loads ranging from 105 to 109 CFU/g, including the cheese samples where no L. lactis was found by RT-qPCR. From these cheeses, none of the colonies isolated on M17 medium was identified as L. lactis species. These data could be interpreted as a lack of selectivity of M17 medium where colony growth is not always related to lactococcal species. At the same time, the absence or low abundance of L. lactis isolates on M17 medium from cheese where L. lactis was detected by RT-qPCR support the hypothesis that L. lactis starter populations are mainly present in viable but not culturable state during ripening and, for this reason, culture-dependent methods have to be supplemented with direct analysis of cheese. PMID:25503474

  17. Methods for assessing pre-induction cervical ripening

    PubMed Central

    Ezebialu, Ifeanyichukwu U; Eke, Ahizechukwu C; Eleje, George U; Nwachukwu, Chukwuemeka E

    2015-01-01

    Background Induction of labour is the artificial initiation of labour in a pregnant woman after the age of fetal viability but without any objective evidence of active phase labour and with intact fetal membranes. The need for induction of labour may arise due to a problem in the mother, her fetus or both, and the procedure may be carried out at or before term. Obstetricians have long known that for this to be successful, it is important that the uterine cervix (the neck of the womb) has favourable characteristics in terms of readiness to go into the labour state. Objectives To compare Bishop score with any other method for assessing pre-induction cervical ripening in women admitted for induction of labour. Search methods We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (31 March 2015) and reference lists of retrieved studies to identify randomised controlled trials (RCTs). Selection criteria All RCTs comparing Bishop score with any other methods of pre-induction cervical assessment in women admitted for induction of labour. Cluster-RCTs were eligible for inclusion but none were identified. Quasi-RCTs and studies using a cross-over design were not eligible for inclusion. Studies published in abstract form were eligible for inclusion if they provided sufficient information. Comparisons could include the following. Bishop score versus transvaginal ultrasound (TVUS). Bishop score versus Insulin-like growth factor binding protein-1 (IGFBP-1). Bishop score versus vaginal fetal fibronectin (fFN). However, we only identified data for a comparison of Bishop score versus TVUS. Data collection and analysis Two review authors independently assessed the trials for inclusion, extracted the data and assessed trial quality. Data were checked for accuracy. Main results We included two trials that recruited a total of 234 women. The overall risk of bias was low for the two studies. Both studies compared Bishop score withTVUS. The two included studies did

  18. Changes in proteins during the ripening of Spanish dried beef 'Cecina'.

    PubMed

    García, I; Díez, V; Zumalacárregui, J M

    1997-08-01

    Changes in the solubility of sarcoplasmic and myofibrillar proteins were tracked in Semitendinosus and Rectus femoris muscles during the ripening process of Spanish 'Cecina'. The extractability of both types of proteins decreased during the ripening. This phenomenon was more marked in the initial stages of processing. Electrophoretic studies of the myofibrillar proteins showed the virtual disappearance of the myosin heavy chain, troponin C and myosin light chain 2 from the smoking phase onward and the appearance of three components of molecular weight of about 65, 70 and 75 kda during ripening. The remaining proteins did not suffer appreciable changes.

  19. Delayed ripening of banana fruit by salicylic acid.

    PubMed

    Srivastava; Dwivedi

    2000-09-08

    Salicylic acid treatment has been found to delay the ripening of banana fruits (Musa acuminata). Fruit softening, pulp:peel ratio, reducing sugar content, invertase and respiration rate have been found to decrease in salicylic acid treated fruits as compared with control ones. The activities of major cell wall degrading enzymes, viz. cellulase, polygalacturonase and xylanase were found to be decreased in presence of salicylic acid. The major enzymatic antioxidants namely, catalase and peroxidase, were also found to be decreased in presence of salicylic acid during banana fruit ripening.

  20. 2D-DIGE analysis of mango (Mangifera indica L.) fruit reveals major proteomic changes associated with ripening.

    PubMed

    Andrade, Jonathan de Magalhães; Toledo, Tatiana Torres; Nogueira, Silvia Beserra; Cordenunsi, Beatriz Rosana; Lajolo, Franco Maria; do Nascimento, João Roberto Oliveira

    2012-06-18

    A comparative proteomic investigation between the pre-climacteric and climacteric mango fruits (cv. Keitt) was performed to identify protein species with variable abundance during ripening. Proteins were phenol-extracted from fruits, cyanine-dye-labeled, and separated on 2D gels at pH 4-7. Total spot count of about 373 proteins spots was detected in each gel and forty-seven were consistently different between pre-climacteric and climacteric fruits and were subjected to LC-MS/MS analysis. Functional classification revealed that protein species involved in carbon fixation and hormone biosynthesis decreased during ripening, whereas those related to catabolism and the stress-response, including oxidative stress and abiotic and pathogen defense factors, accumulated. In relation to fruit quality, protein species putatively involved in color development and pulp softening were also identified. This study on mango proteomics provides an overview of the biological processes that occur during ripening. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Overexpression of a novel MADS-box gene SlFYFL delays senescence, fruit ripening and abscission in tomato

    PubMed Central

    Xie, Qiaoli; Hu, Zongli; Zhu, Zhiguo; Dong, Tingting; Zhao, Zhiping; Cui, Baolu; Chen, Guoping

    2014-01-01

    MADS-domain proteins are important transcription factors involved in many biological processes of plants. In our study, a tomato MADS-box gene, SlFYFL, was isolated. SlFYFL is expressed in all tissues of tomato and significantly higher in mature leave, fruit of different stages, AZ (abscission zone) and sepal. Delayed leaf senescence and fruit ripening, increased storability and longer sepals were observed in 35S:FYFL tomato. The accumulation of carotenoid was reduced, and ethylene content, ethylene biosynthetic and responsive genes were down-regulated in 35S:FYFL fruits. Abscission zone (AZ) did not form normally and abscission zone development related genes were declined in AZs of 35S:FYFL plants. Yeast two-hybrid assay revealed that SlFYFL protein could interact with SlMADS-RIN, SlMADS1 and SlJOINTLESS, respectively. These results suggest that overexpression of SlFYFL regulate fruit ripening and development of AZ via interactions with the ripening and abscission zone-related MADS box proteins. PMID:24621662

  2. Inhibitor development after liver transplantation in congenital factor VII deficiency.

    PubMed

    See, W-S Q; Chang, K-O; Cheuk, D K-L; Leung, Y-Y R; Chan, G C-F; Chan, S-C; Ha, S-Y

    2016-09-01

    Congenital factor VII (FVII) deficiency is the commonest type of the rare bleeding disorders. Very few cases of congenital FVII deficiency developed inhibitor and liver transplant is considered as definitive treatment. In the literature, twelve patients with congenital FVII deficiency developed inhibitors. Two had spontaneous resolution of inhibitors and one did not respond to high dose recombinant factor VIIa (rFVIIa) and died. Regarding liver transplant in congenital FVII patients, seven patients underwent liver transplant with good prognosis. We report a 5-year-old girl with confirmed severe congenital FVII deficiency since neonatal period. She suffered from recurrent intracranial bleeding despite rFVIIa replacement. After auxiliary liver transplant at the age of 4, she continued to show persistent deranged clotting profile and was found to have inhibitor towards FVII. Interestingly, she was still responsive to rFVIIa replacement. © 2016 John Wiley & Sons Ltd.

  3. Recombinant factor concentrates may increase inhibitor development: a single centre cohort study.

    PubMed

    Strauss, T; Lubetsky, A; Ravid, B; Bashari, D; Luboshitz, J; Lalezari, S; Misgav, M; Martinowitz, U; Kenet, G

    2011-07-01

    Recent reports have raised concerns regarding potential risk factors for inhibitor development. In Israel, all haemophilia patients (n = 479) are followed by the National Hemophilia Center. Most children are neonatally exposed to factor concentrate (due to circumcision performed at the age of 8 days). The impact of early exposure and recombinant FVIII products (rFVIII) administration (approved in Israel since 1996) upon inhibitor occurrence in our cohort of haemophilia A (HA) patients was analysed. Two hundred ninety-two consecutive paediatric cases with a first symptomatic onset of HA were enrolled and followed over a median time of 7 years [min-max: 9 months to 17 years]. Study endpoint was inhibitor development against factor VIII. In addition, the treatment regimens applied, i.e. bolus administration or 'continuous infusion' and the family history of inhibitor development were investigated. During the follow-up period 31/292 children (10.6%) developed high titre inhibitors. Inhibitors occurred in 14/43 (32.5%) HA patients neonatally exposed to rFVIII, as compared to 22/249 previously treated with Plasma Derived (PD) products (8.8%). The odds ratio for inhibitor formation in rFVIII treated HA patients was 3.43 (95% CI: 1.36-8.65). Transient inhibitor evolved among 2/43 paediatric HA patients, only among those treated with rFVIII. The risk of inhibitor detection significantly increased among HA children treated by continuous infusion (P = 0.025). Our experience shows that the risk of inhibitor formation may be increased by early exposure to recombinant concentrates. The multiple variables affecting inhibitor incidence deserve further attention by larger prospective studies. © 2011 Blackwell Publishing Ltd.

  4. A hydrothermal after-ripening time model for seed dormancy loss in Bromus tectorum L.

    Treesearch

    Necia B. Bair; Susan E. Meyer; Phil S. Allen

    2006-01-01

    After-ripening, the loss of dormancy under dry conditions, is associated with a decrease in mean base water potential for germination of Bromus tectorum L. seeds. After-ripening rate is a linear function of temperature above a base temperature, so that dormancy loss can be quantified using a thermal after-ripening time (TAR) model. To incorporate storage water...

  5. Analysis of strawberry ripening by dynamic speckle measurements

    NASA Astrophysics Data System (ADS)

    Mulone, C.; Budini, N.; Vincitorio, F. M.; Freyre, C.; López Díaz, A. J.; Ramil Rego, A.

    2013-11-01

    This work seeks to determine the age of a fruit from observation of its dynamic speckle pattern. A mobile speckle pattern originates on the fruit's surface due to the interference of the wavefronts reflected from moving scatterers. For this work we analyzed two series of photographs of a strawberry speckle pattern, at different stages of ripening, acquired with a CMOS camera. The first day, we took ten photographs at an interval of one second. The same procedure was repeated the next day. From each series of images we extracted several statistical descriptors of pixel-to-pixel gray level variation during the observation time. By comparing these values from the first to the second day we noticed a diminution of the speckle activity. This decay demonstrated that after only one day the ripening process of the strawberry can be detected by dynamic speckle pattern analysis. For this study we employed a simple new algorithm to process the data obtained from the photographs. This algorithm allows defining a global mobility index that indicates the evolution of the fruit's ripening.

  6. [Subchronic toxicity testing of mold-ripened cheese].

    PubMed

    Schoch, U; Lüthy, J; Schlatter, C

    1984-08-01

    The biological effects of known mycotoxins of Penicillium roqueforti or P. camemberti and other still unknown, but potentially toxic metabolites in mould ripened cheese (commercial samples of Blue- and Camembert cheese) were investigated. High amounts of mycelium (equivalents of 100 kg cheese/man and day) were fed to mice in a subchronic feeding trial. The following parameters were determined: development of body weight, organ weights, hematology, blood plasma enzymes. No signs of adverse effects produced by cheese mycotoxins could be detected after 28 days. No still unknown toxic metabolites could be demonstrated. From these results no health hazard from the consumption of mould ripened cheese, even in high amounts, appears to exist.

  7. Evolution of ellagitannin content and profile during fruit ripening in Fragaria spp.

    PubMed

    Gasperotti, Mattia; Masuero, Domenico; Guella, Graziano; Palmieri, Luisa; Martinatti, Paolo; Pojer, Elisa; Mattivi, Fulvio; Vrhovsek, Urska

    2013-09-11

    Ellagitannins and ellagic acid conjugates are polyphenols present in the human diet, in particular strawberries (Fragaria spp.). The first aim of this study was isolation and structural characterization of casuarictin and 3-O-methyl ellagic acid 3'-O-α-rhamnopyranoside, which were found to be abundant in Fragaria spp., along with agrimoniin. The second aim was accurate profiling and quantification of 26 ellagitannins and ellagic acid conjugates in six Fragaria x ananassa cultivars and two Fragaria vesca species. The third aim was to describe the ellagitannins behavior during fruit ripening from the green stage to over-ripeness. It was shown that there are major qualitative and quantitative differences in the amount and profile of ellagitannins and ellagic acid conjugates between Fragaria spp. Genotype is a major factor in defining ellagitannin concentration and patterns between strawberries, and variable behavior of the genotypes was observed, in the context of a significant drop in ellagitannins during ripening.

  8. Hemodialysis in a patient with severe hemophilia A and factor VIII inhibitor.

    PubMed

    Gopalakrishnan, Natarajan; Usha, Thiruvengadam; Thopalan, Balasubramaniyan; Dhanapriya, Jeyachandran; Dineshkumar, Thanigachalam; Thirumalvalavan, Kaliaperumal; Sakthirajan, Ramanathan

    2016-10-01

    Hemophilia A is a hereditary X-linked recessive disease caused by mutations in the gene encoding factor VIII (FVIII), occurring in 1 out of 10,000 persons. Life expectancy and quality of life have dramatically improved recently in patients with hemophilia. Chronic kidney disease and need for renal replacement therapy in these patients are rare. The development of inhibitors to FVIII is the most serious complication of hemophilia and makes treatment of bleeds very challenging. We describe here a 28-year-old male patient with severe hemophilia A with presence of factor VIII inhibitor, who had end stage renal disease. Central venous access device was inserted along with infusion of factor eight inhibitor bypass activity before and after the procedure. He is currently on thrice weekly hemodialysis and doing well for 6 months without bleeding episodes. To our knowledge, hemophilia A with factor VIII inhibitor managed with hemodialysis has not been reported so far. © 2016 International Society for Hemodialysis.

  9. Kinetic synergistic transitions in the Ostwald ripening processes

    NASA Astrophysics Data System (ADS)

    Sachkov, I. N.; Turygina, V. F.; Dolganov, A. N.

    2018-01-01

    There is proposed approach to mathematical description of the kinetic transitions in Ostwald ripening processes of volatile substance in nonuniformly heated porous materials. It is based upon the finite element method. There are implemented computer software. The main feature of the software is to calculate evaporation and condensation fluxes on the walls of a nonuniformly heated cylindrical capillary. Kinetic transitions are detected for three modes of volatile substances migration which are different by condensation zones location. There are controlling dimensionless parameters of the kinetic transition which are revealed during research. There is phase diagram of the Ostwald ripening process modes realization.

  10. Von Willebrand factor-containing factor VIII concentrates and inhibitors in haemophilia A. A critical literature review.

    PubMed

    Franchini, Massimo; Lippi, Giuseppe

    2010-11-01

    The development of inhibitors that neutralise the function of factor VIII (FVIII) is currently not only the most challenging complication associated with the treatment of haemophilia A but it also increases the disease-related morbidity as bleeding episodes do not respond to standard therapy. The main short-term goal of the treatment of inhibitor patients is to control bleeding episodes while the long-term one is to permanently eradicate the inhibitor by immune tolerance induction, particularly in the case of high-titer antibodies. Due to some in vitro studies and clinical observations, some investigators have suggested that FVIII concentrates containing von Willebrand factor (VWF) may be less immunogenic than high-purity or recombinant FVIII products. It has also been suggested that success rates for immune tolerance induction are higher when plasma-derived FVIII products are used. The currently available data from laboratory and clinical studies on the role of VWF in inhibitor development and eradication in haemophilia A is critically analysed in this review. As a result, we have not found definitive evidence supporting a role for product type on inhibitor incidence and inhibitor eradication in haemophilia A patients.

  11. Small molecule inhibitors of anthrax edema factor.

    PubMed

    Jiao, Guan-Sheng; Kim, Seongjin; Moayeri, Mahtab; Thai, April; Cregar-Hernandez, Lynne; McKasson, Linda; O'Malley, Sean; Leppla, Stephen H; Johnson, Alan T

    2018-01-15

    Anthrax is a highly lethal disease caused by the Gram-(+) bacteria Bacillus anthracis. Edema toxin (ET) is a major contributor to the pathogenesis of disease in humans exposed to B. anthracis. ET is a bipartite toxin composed of two proteins secreted by the vegetative bacteria, edema factor (EF) and protective antigen (PA). Our work towards identifying a small molecule inhibitor of anthrax edema factor is the subject of this letter. First we demonstrate that the small molecule probe 5'-Fluorosulfonylbenzoyl 5'-adenosine (FSBA) reacts irreversibly with EF and blocks enzymatic activity. We then show that the adenosine portion of FSBA can be replaced to provide more drug-like molecules which are up to 1000-fold more potent against EF relative to FSBA, display low cross reactivity when tested against a panel of kinases, and are nanomolar inhibitors of EF in a cell-based assay of cAMP production. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Ostwald ripening theory

    NASA Technical Reports Server (NTRS)

    Baird, J. K.

    1986-01-01

    The Ostwald-ripening theory is deduced and discussed starting from the fundamental principles such as Ising model concept, Mayer cluster expansion, Langer condensation point theory, Ginzburg-Landau free energy, Stillinger cutoff-pair potential, LSW-theory and MLSW-theory. Mathematical intricacies are reduced to an understanding version. Comparison of selected works, from 1949 to 1984, on solution of diffusion equation with and without sink/sources term(s) is presented. Kahlweit's 1980 work and Marqusee-Ross' 1954 work are more emphasized. Odijk and Lekkerkerker's 1985 work on rodlike macromolecules is introduced in order to simulate interested investigators.

  13. Influence of calcium and phosphorus, lactose, and salt-to-moisture ratio on Cheddar cheese quality: changes in residual sugars and water-soluble organic acids during ripening.

    PubMed

    Upreti, P; McKay, L L; Metzger, L E

    2006-02-01

    Cheddar cheese ripening involves the conversion of lactose to glucose and galactose or galactose-6-phosphate by starter and nonstarter lactic acid bacteria. Under ideal conditions (i.e., where bacteria grow under no stress of pH, water activity, and salt), these sugars are mainly converted to lactic acid. However, during ripening of cheese, survival and growth of bacteria occurs under the stressed condition of low pH, low water activity, and high salt content. This forces bacteria to use alternate biochemical pathways resulting in production of other organic acids. The objective of this study was to determine if the level and type of organic acids produced during ripening was influenced by calcium (Ca) and phosphorus (P), residual lactose, and salt-to-moisture ratio (S/M) of cheese. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), lactose at pressing (2.4 vs. 0.78%), and S/M (6.4 vs. 4.8%) were manufactured. The cheeses were analyzed for organic acids (citric, orotic, pyruvic, lactic, formic, uric, acetic, propanoic, and butyric acids) and residual sugars (lactose, galactose) during 48 wk of ripening using an HPLC-based method. Different factors influenced changes in concentration of residual sugars and organic acids during ripening and are discussed in detail. Our results indicated that the largest decrease in lactose and the largest increase in lactic acid occurred between salting and d 1 of ripening. It was interesting to observe that although the lactose content in cheese was influenced by several factors (Ca and P, residual lactose, and S/M), the concentration of lactic acid was influenced only by S/M. More lactic acid was produced in low S/M treatments compared with high S/M treatments. Although surprising for Cheddar cheese, a substantial amount (0.2 to 0.4%) of galactose was observed throughout ripening in all treatments. Minor changes in the levels of citric, uric, butyric, and propanoic acids were observed during

  14. Microbiology, biochemistry, and volatile composition of Tulum cheese ripened in goat's skin or plastic bags.

    PubMed

    Hayaloglu, A A; Cakmakci, S; Brechany, E Y; Deegan, K C; McSweeney, P L H

    2007-03-01

    Tulum cheeses were manufactured from raw ewe's milk and ripened in goat's skin bags (tulums) or plastic containers to understand the effect of ripening container on the chemical composition, biochemistry, microbiology, and volatile composition of Tulum cheeses during 150 d of ripening. Chemical compositions of the cheeses ripened in tulums were significantly different and the moisture contents decreased rapidly in those cheeses because of the porous structure of the tulum. Higher microbial counts were detected in the cheeses ripened in plastic than in cheeses ripened in tulums. Differences in nitrogenous compounds and total free AA of the cheeses were not significant. Total concentrations of free AA in cheeses increased with age and Glu, Ala, Val, Leu, and Phe were the most abundant AA in the cheeses. Urea-PAGE of pH 4.6-insoluble fractions of the cheeses during ripening showed similar degradation patterns in all cheeses. Peptide profiles by reversed-phase HPLC of pH 4.6- and ethanol-soluble or ethanol-insoluble fractions of the cheeses revealed only minor differences in the concentrations of some peptides among the cheeses; however, age-related changes in peptide concentrations were significantly different among the cheeses. Cheeses were analyzed at 90 d of ripening for volatile compounds by solid-phase microextraction gas chromatography-mass spectrometry. One hundred volatile components were identified, including 11 acids, 16 esters, 12 methyl ketones, 7 aldehydes, 22 alcohols, 7 sulfur compounds, 6 terpenes, and 19 miscellaneous compounds. The main components were short-chain fatty acids, 2-butanone, diacetyl, and primary alcohols. Quantitative differences in several volatile compounds were evident among the cheeses. Cheeses ripened in tulums or plastic had similar aroma patterns, but the concentrations of some components were different.

  15. Artificial Ripening of Sweetgum Seeds

    Treesearch

    F. T. Bonner

    1969-01-01

    Untrained collectors of hardwood seeds often start picking the seeds too early in the year. The immature seeds germinate poorly, if at all, and nursery production suffers. In the study reported here, sweetgum (Liquidambar styraciflua L.) seeds collected in central Mississippi in mid-August were ripe. Seeds collected as early as July 19 were artificially ripened by...

  16. Evaluation of chemical parameters in soft mold-ripened cheese during ripening by mid-infrared spectroscopy.

    PubMed

    Martín-del-Campo, S T; Picque, D; Cosío-Ramírez, R; Corrieu, G

    2007-06-01

    The suitability of mid-infrared spectroscopy (MIR) to follow the evolution throughout ripening of specific physicochemical parameters in Camembert-type cheeses was evaluated. The infrared spectra were obtained directly from raw cheese samples deposited on an attenuated total reflectance crystal. Significant correlations were observed between physicochemical data, pH, acid-soluble nitrogen, nonprotein nitrogen, ammonia (NH4+), lactose, and lactic acid. Dry matter showed significant correlation only with lactose and nonprotein nitrogen. Principal components analysis factorial maps of physicochemical data showed a ripening evolution in 2 steps, from d 1 to d 7 and from d 8 to d 27, similar to that observed previously from infrared spectral data. Partial least squares regressions made it possible to obtain good prediction models for dry matter, acid-soluble nitrogen, nonprotein nitrogen, lactose, lactic acid, and NH4+ values from spectral data of raw cheese. The values of 3 statistical parameters (coefficient of determination, root mean square error of cross validation, and ratio prediction deviation) are satisfactory. Less precise models were obtained for pH.

  17. Gr and hp-1 tomato mutants unveil unprecedented interactions between arbuscular mycorrhizal symbiosis and fruit ripening.

    PubMed

    Chialva, Matteo; Zouari, Inès; Salvioli, Alessandra; Novero, Mara; Vrebalov, Julia; Giovannoni, James J; Bonfante, Paola

    2016-07-01

    Systemic responses to an arbuscular mycorrhizal fungus reveal opposite phenological patterns in two tomato ripening mutants depending whether ethylene or light reception is involved. The availability of tomato ripening mutants has revealed many aspects of the genetics behind fleshy fruit ripening, plant hormones and light signal reception. Since previous analyses revealed that arbuscular mycorrhizal symbiosis influences tomato berry ripening, we wanted to test the hypothesis that an interplay might occur between root symbiosis and fruit ripening. With this aim, we screened seven tomato mutants affected in the ripening process for their responsiveness to the arbuscular mycorrhizal fungus Funneliformis mosseae. Following their phenological responses we selected two mutants for a deeper analysis: Green ripe (Gr), deficient in fruit ethylene perception and high-pigment-1 (hp-1), displaying enhanced light signal perception throughout the plant. We investigated the putative interactions between ripening processes, mycorrhizal establishment and systemic effects using biochemical and gene expression tools. Our experiments showed that both mutants, notwithstanding a normal mycorrhizal phenotype at root level, exhibit altered arbuscule functionality. Furthermore, in contrast to wild type, mycorrhization did not lead to a higher phosphate concentration in berries of both mutants. These results suggest that the mutations considered interfere with arbuscular mycorrhiza inducing systemic changes in plant phenology and fruits metabolism. We hypothesize a cross talk mechanism between AM and ripening processes that involves genes related to ethylene and light signaling.

  18. Expression patterns of ethylene biosynthesis genes from bananas during fruit ripening and in relationship with finger drop

    PubMed Central

    Hubert, Olivier; Mbéguié-A-Mbéguié, Didier

    2012-01-01

    Background and aims Banana finger drop is defined as dislodgement of individual fruits from the hand at the pedicel rupture area. For some banana varieties, this is a major feature of the ripening process, in addition to ethylene production and sugar metabolism. The few studies devoted to assessing the physiological and molecular basis of this process revealed (i) the similarity between this process and softening, (ii) the early onset of related molecular events, between the first and fourth day after ripening induction, and (iii) the putative involvement of ethylene as a regulatory factor. This study was conducted with the aim of identifying, through a candidate gene approach, a quality-related marker that could be used as a tool in breeding programmes. Here we examined the relationship between ripening ethylene biosynthesis (EB) and finger drop in order to gain further insight into the upstream regulatory steps of the banana finger drop process and to identify putative related candidate genes. Methods Postharvest ripening of green banana fruit was induced by acetylene treatment and fruit taken at 1–4 days after ripening induction, and total RNA extracted from the median area [control zone (CZ)] and the pedicel rupture area [drop zone (DZ)] of peel tissue. Then the expression patterns of EB genes (MaACO1, MaACO2, MaACS1, MaACS2, MaACS3 and MaACS4) were comparatively examined in CZ and DZ via real-time quantitative polymerase chain reaction. Principal results Differential expression of EB gene was observed in CZ and DZ during the postharvest period examined in this study. MaACO1, MaACS2 and MaACS1 were more highly induced in DZ than in the control, while a slight induction of the MaACS4 gene was observed. No marked differences between the two zones were observed for the MaACO2 gene. Conclusions The finger drop process enhanced EB gene expression including developmental- and ripening-induced genes (MaACO1), specific ripening-induced genes (MaACS1) and wound

  19. Dynamic changes in the date palm fruit proteome during development and ripening

    PubMed Central

    Marondedze, Claudius; Gehring, Christoph; Thomas, Ludivine

    2014-01-01

    Date palm (Phoenix dactylifera) is an economically important fruit tree in the Middle East and North Africa and is characterized by large cultivar diversity, making it a good model for studies on fruit development and other important traits. Here in gel comparative proteomics combined with tandem mass spectrometry were used to study date fruit development and ripening. Total proteins were extracted using a phenol-based protocol. A total of 189 protein spots were differentially regulated (p≤0.05). The identified proteins were classified into 14 functional categories. The categories with the most proteins were ‘disease and defense’ (16.5%) and ‘metabolism’ (15.4%). Twenty-nine proteins have not previously been identified in other fleshy fruits and 64 showed contrasting expression patterns in other fruits. Abundance of most proteins with a role in abiotic stress responses increased during ripening with the exception of heat shock proteins. Proteins with a role in anthocyanin biosynthesis, glycolysis, tricarboxylic acid cycle and cell wall degradation were upregulated particularly from the onset of ripening and during ripening. In contrast, expression of pentose phosphate- and photosynthesis-related proteins decreased during fruit maturation. Although date palm is considered a climacteric species, the analysis revealed downregulation of two enzymes involved in ethylene biosynthesis, suggesting an ethylene-independent ripening of ‘Barhi’ fruits. In summary, this proteomics study provides insights into physiological processes during date fruit development and ripening at the systems level and offers a reference proteome for the study of regulatory mechanisms that can inform molecular and biotechnological approaches to further improvements of horticultural traits including fruit quality and yield. PMID:26504545

  20. Oral direct thrombin inhibitors or oral factor Xa inhibitors for the treatment of deep vein thrombosis.

    PubMed

    Robertson, Lindsay; Kesteven, Patrick; McCaslin, James E

    2015-06-30

    Deep vein thrombosis (DVT) is a condition in which a clot forms in the deep veins, most commonly of the leg. It occurs in approximately 1 in 1,000 people. If left untreated, the clot can travel up to the lungs and cause a potentially life-threatening pulmonary embolism (PE). Previously, a DVT was treated with the anticoagulants heparin and vitamin K antagonists. However, two forms of novel oral anticoagulants (NOACs) have been developed: oral direct thrombin inhibitors (DTI) and oral factor Xa inhibitors. The new drugs have characteristics that may be favourable over conventional treatment, including oral administration, a predictable effect, lack of frequent monitoring or re-dosing and few known drug interactions. To date, no Cochrane review has measured the effectiveness and safety of these drugs in the treatment of DVT. To assess the effectiveness of oral DTIs and oral factor Xa inhibitors for the treatment of DVT. The Cochrane Peripheral Vascular Diseases Group Trials Search Co-ordinator searched the Specialised Register (last searched January 2015) and the Cochrane Register of Studies (last searched January 2015). We searched clinical trials databases for details of ongoing or unpublished studies and the reference lists of relevant articles retrieved by electronic searches for additional citations. We included randomised controlled trials in which people with a DVT confirmed by standard imaging techniques, were allocated to receive an oral DTI or an oral factor Xa inhibitor for the treatment of DVT. Two review authors (LR, JM) independently extracted the data and assessed the risk of bias in the trials. Any disagreements were resolved by discussion with the third review author (PK). We performed meta-analyses when we considered heterogeneity low. The two primary outcomes were recurrent VTE and PE. Other outcomes included all-cause mortality and major bleeding. We calculated all outcomes using an odds ratio (OR) with a 95% confidence interval (CI). We included

  1. Influence of ripening stages on antioxidant properties of papaya fruit (Carica papaya L.)

    NASA Astrophysics Data System (ADS)

    Addai, Zuhair Radhi; Abdullah, Aminah; Mutalib, Sahilah Abd.

    2013-11-01

    Papaya (Carica papaya L. cv Eksotika) is one of the most commonly consumed tropical fruits by humans, especially Malaysians. The objective of this study was to determine the phenolic compounds and antioxidants activity in different ripening stages of papaya fruit. The fruits were harvested at five different, stages RS1, RS2, RS3, RS4, and RS5 corresponding to 12, 14, 16, 18, and 20 weeks after anthesis, respectively. Papayas fruit at five different stage of ripening were obtained from farms at Pusat Flora Cheras, JabatanPertanian and Hulu Langat Semenyih, Selangor, Malaysia. The antioxidants activity were analyzed using the total phenolic content (TPC), total flavonoid content (TFC), ferric reducing antioxidant Power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH). The analyses were conducted in triplicate and the data were subjected to statistical analysis using SPSS. The results showed significant differences (P< 0.05) were found at different stages of ripening. The total phenol content TPC, TFC, FRAP and DPPH values increased significantly (P<0.05) with the ripening process. The results showed the important role of the ripening stage in increasing the antioxidant content of papaya fruits.

  2. How chemical ripeners of sugarcane affect the starch and color quality of juices

    USDA-ARS?s Scientific Manuscript database

    The application of chemical ripeners is an important component of sugarcane cultivation management to increase sucrose concentrations. Unfortunately, very little information is available concerning the effects of ripener on quality parameters which are critical to both factory and refinery processi...

  3. Temporal Sequence of Cell Wall Disassembly in Rapidly Ripening Melon Fruit1

    PubMed Central

    Rose, Jocelyn K.C.; Hadfield, Kristen A.; Labavitch, John M.; Bennett, Alan B.

    1998-01-01

    The Charentais variety of melon (Cucumis melo cv Reticulatus F1 Alpha) was observed to undergo very rapid ripening, with the transition from the preripe to overripe stage occurring within 24 to 48 h. During this time, the flesh first softened and then exhibited substantial disintegration, suggesting that Charentais may represent a useful model system to examine the temporal sequence of changes in cell wall composition that typically take place in softening fruit. The total amount of pectin in the cell wall showed little reduction during ripening but its solubility changed substantially. Initial changes in pectin solubility coincided with a loss of galactose from tightly bound pectins, but preceded the expression of polygalacturonase (PG) mRNAs, suggesting early, PG-independent modification of pectin structure. Depolymerization of polyuronides occurred predominantly in the later ripening stages, and after the appearance of PG mRNAs, suggesting the existence of PG-dependent pectin degradation in later stages. Depolymerization of hemicelluloses was observed throughout ripening, and degradation of a tightly bound xyloglucan fraction was detected at the early onset of softening. Thus, metabolism of xyloglucan that may be closely associated with cellulose microfibrils may contribute to the initial stages of fruit softening. A model is presented of the temporal sequence of cell wall changes during cell wall disassembly in ripening Charentais melon. PMID:9625688

  4. Activation of ethylene-responsive p-hydroxyphenylpyruvate dioxygenase leads to increased tocopherol levels during ripening in mango

    PubMed Central

    Singh, Rajesh K.; Ali, Sharique A.; Nath, Pravendra; Sane, Vidhu A.

    2011-01-01

    Mango is characterized by high tocopherol and carotenoid content during ripening. From a cDNA screen of differentially expressing genes during mango ripening, a full-length p-hydroxyphenylpyruvate dioxygenase (MiHPPD) gene homologue was isolated that encodes a key enzyme in the biosynthesis of tocopherols. The gene encoded a 432-amino-acid protein. Transcript analysis during different stages of ripening revealed that the gene is ripening related and rapidly induced by ethylene. The increase in MiHPPD transcript accumulation was followed by an increase in tocopherol levels during ripening. The ripening-related increase in MiHPPD expression was also seen in response to abscisic acid and to alesser extent to indole-3-acetic acid. The expression of MiHPPD was not restricted to fruits but was also seen in other tissues such as leaves particularly during senescence. The strong ethylene induction of MiHPPD was also seen in young leaves indicating that ethylene induction of MiHPPD is tissue independent. Promoter analysis of MiHPPD gene in tomato discs and leaves of stable transgenic lines of Arabidopsis showed that the cis elements for ripening-related, ethylene-responsive, and senescence-related expression resided within the 1590 nt region upstream of the ATG codon. Functionality of the gene was demonstrated by the ability of the expressed protein in bacteria to convert p-hydroxyphenylpyruvate to homogentisate. These results provide the first evidence for HPPD expression during ripening of a climacteric fruit. PMID:21430290

  5. Impact of vitamin E and selenium on antioxidant capacity and lipid oxidation of cheddar cheese in accelerated ripening.

    PubMed

    Batool, Maryam; Nadeem, Muhammad; Imran, Muhammad; Gulzar, Nabila; Shahid, Muhammad Qamar; Shahbaz, Muhammad; Ajmal, Muhammad; Khan, Imran Taj

    2018-04-11

    Ripening of cheddar cheese is a time taking process, duration of the ripening may be as long as one year. Long ripening time is a big hindrance in the popularity of cheese in developing countries. Further, energy resources in these countries are either insufficient or very expensive. Therefore, those methods of cheese ripening should be discovered which can significantly reduce the ripening time without compromising the quality characteristics of cheddar cheese. In accelerated ripening, cheese is usually ripened at higher temperature than traditional ripening temperatures. Ripening of cheddar cheese at high temperature with the addition of vitamin E and selenium is not previously studied. This investigation aimed to study the antioxidant activity of selenium and vitamin E in accelerated ripening using cheddar cheese as an oxidation substrate. The ripening of cheddar cheese was performed at 18 °C and to prevent lipid oxidation, vitamin E and selenium were used alone and in combination. The treatments were as: cheddar cheese without any addition of vitamin E and selenium (T1), cheddar cheese added with 100 mg/kg vitamin E (T 2 ), 200 mg/kg vitamin E (T 3 ), 800 μg/kg selenium (T 4 ), 1200 μg/kg selenium (T 5 ), vitamin E 100 mg/kg + 800 μg/kg selenium (T 6 ) and vitamin E 200 mg/kg + 1200 μg/kg selenium (T 7 ). Traditional cheddar cheese ripne ripened at 4-6 °C for 9 months was used as positive control. Cheese samples were ripened at 18 °C for a period of 12 weeks and analyzed for chemical and oxidative stability characteristics at 0, 6 and 12 weeks of storage. All these treatments were compared with a cheddar cheese without vitamin E, selenium and ripened at 4 °C or 12 weeks. Vacuum packaged cheddar cheese was ripened 18 °C for a period of 12 weeks and analyzed for chemical and oxidative stability characteristics at 0, 4 and 8 weeks of storage period. Addition of Vitamin E and selenium did not have any effect on moisture, fat and

  6. Carbohydrate Analysis: Can We Control the Ripening of Bananas?

    NASA Astrophysics Data System (ADS)

    Deal, S. Todd; Farmer, Catherine E.; Cerpovicz, Paul F.

    2002-04-01

    We have developed an experiment for nutritional/introductory biochemistry courses that focuses on carbohydrate analysis--specifically, the carbohydrates found in bananas and the change in carbohydrate composition as the banana ripens. Pairs of students analyze the starch and reducing sugar content of green, ripe, and overripe bananas. Using the techniques and knowledge gained from these analyses, they then investigate the influence of various storage methods on the ripening process. While this experiment was developed for an introductory-level biochemistry lab, it can easily be adapted for use in other laboratory programs that seek to teach the fundamentals of carbohydrate analysis.

  7. Anticoagulation by factor Xa inhibitors.

    PubMed

    Orfeo, T; Butenas, S; Brummel-Ziedins, K E; Gissel, M; Mann, K G

    2010-08-01

    Therapeutic agents that regulate blood coagulation are critical to the management of thrombotic disorders, with the selective targeting of factor (F) Xa emerging as a promising approach. To assess anticoagulant strategies targeting FXa. A deterministic computational model of tissue factor (Tf)-initiated thrombin generation and two empirical experimental systems (a synthetic coagulation proteome reconstruction using purified proteins and a whole blood model) were used to evaluate clinically relevant examples of the two available types of FXa-directed anticoagulants [an antithrombin (AT)-dependent agent, fondaparinux, and an AT-independent inhibitor, Rivaroxaban] in experimental regimens relevant to long-term (suppression of new Tf-initiated events) and acute (suppression of ongoing coagulation processes) clinical applications. Computational representations of each anticoagulant's efficacy in suppressing thrombin generation over a range of anticoagulant concentrations in both anticoagulation regimens were validated by results from corresponding empirical reconstructions and were consistent with those recommended for long-term and acute clinical applications, respectively. All three model systems suggested that Rivaroxaban would prove more effective in the suppression of an ongoing coagulation process than fondaparinux, reflecting its much higher reactivity toward the prothrombinase complex. The success of fondaparinux in acute settings in vivo is not explained solely by its properties as an FXa inhibitor. We have reported that FIXa contributes to the long-term capacity of clot-associated catalysts to restart a coagulation process, suggesting that the enhanced anti-FIXa activity of fondaparinux-AT may be critical to its success in acute settings in vivo. © 2010 International Society on Thrombosis and Haemostasis.

  8. Theory of Ostwald ripening in a two-component system

    NASA Technical Reports Server (NTRS)

    Baird, J. K.; Lee, L. K.; Frazier, D. O.; Naumann, R. J.

    1986-01-01

    When a two-component system is cooled below the minimum temperature for its stability, it separates into two or more immiscible phases. The initial nucleation produces grains (if solid) or droplets (if liquid) of one of the phases dispersed in the other. The dynamics by which these nuclei proceed toward equilibrium is called Ostwald ripening. The dynamics of growth of the droplets depends upon the following factors: (1) The solubility of the droplet depends upon its radius and the interfacial energy between it and the surrounding (continuous) phase. There is a critical radius determined by the supersaturation in the continuous phase. Droplets with radii smaller than critical dissolve, while droplets with radii larger grow. (2) The droplets concentrate one component and reject the other. The rate at which this occurs is assumed to be determined by the interdiffusion of the two components in the continuous phase. (3) The Ostwald ripening is constrained by conservation of mass; e.g., the amount of materials in the droplet phase plus the remaining supersaturation in the continuous phase must equal the supersaturation available at the start. (4) There is a distribution of droplet sizes associated with a mean droplet radius, which grows continuously with time. This distribution function satisfies a continuity equation, which is solved asymptotically by a similarity transformation method.

  9. Genetic risk factors for inhibitors in haemophilia A.

    PubMed

    Bardi, Edit; Astermark, Jan

    2015-02-01

    The current most serious side effect of haemophilia treatment is inhibitor development. Significant progress has been made over the last decades to understand why this complication occurs in some patients and it seems clear that both genetic and non-genetic factors are involved. Several issues however remain to be settled. A review was undertaken to summarise some key findings regarding the current view and available data on genetic markers of potential importance within this area. The causative F8 mutation, together with the HLA class II alleles, plays a pivotal pathophysiological role in inhibitor development. The types of mutation most frequently associated with inhibitors are large deletions, nonsense mutations, inversions, small deletions/insertions without A-runs, splice-site mutations at conserved nucleotides and certain missense mutations. Regarding HLA class II allele, it has been hard to consistently identify risk alleles. Ethnicity has consistently been associated with inhibitor risk, but the causality of this has so far not been resolved. Among immune regulatory molecules, several polymorphic molecules have been suggested to be of importance. Most of these need additional studies and immune system challenges have to be fully evaluated. Inhibitor risk should be further defined, as patients in the future may be offered non-immunogenic treatments. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  10. Formation of early and advanced Maillard reaction products correlates to the ripening of cheese.

    PubMed

    Spanneberg, Robert; Salzwedel, Grit; Glomb, Marcus A

    2012-01-18

    The present study deals with the characterization of the ripening of cheese. A traditional German acid curd cheese was ripened under defined conditions at elevated temperature, and protein and amino acid modifications were investigated. Degree of proteolysis and analysis of early [Amadori compound furosine (6)] and advanced [N(ε)-carboxymethyllysine (4), N(ε)-carboxyethyllysine (5)] Maillard reaction products confirmed the maturation to proceed from the rind to the core of the cheese. Whereas 6 was decreased, 4 and 5 increased over time. Deeper insight into the Maillard reaction during the ripening of cheese was achieved by the determination of selected α-dicarbonyl compounds. Especially methylglyoxal (2) showed a characteristic behavior during storage of the acid curd cheese. Decrease of this reactive structure was directly correlated to the formation of 5. To extend the results of experimental ripening to commercial cheeses, different aged Gouda types were investigated. Maturation times of the samples ranged from 6 to 8 weeks (young) to more than 1 year (aged). Again, increase of 5 and decrease of 2 were able to describe the ripening of this rennet coagulated cheese. Therefore, both chemical parameters are potent markers to characterize the degree of maturation, independent of coagulation.

  11. Report of a factor VIII inhibitor in a patient with autoimmune lymphoproliferative syndrome.

    PubMed

    Fang, B S; Sneller, M C; Straus, S E; Frenkel, L; Dale, J K; Rick, M E

    2000-07-01

    The occurrence of factor VIII inhibitors in non-hemophilic patients is a rare event with a potentially lethal outcome. Despite its infrequent occurrence, the association of this inhibitor with multiple autoimmune diseases is well recognized. We report the case of a patient with the recently described autoimmune lymphoproliferative syndrome (ALPS) who developed an inhibitor to factor VIII. ALPS is a disease characterized by defective lymphocyte apoptosis due to inherited mutations in genes that regulate apoptosis, with the resulting enlargement of lymphoid organs and a variety of autoimmune manifestations. Published 2000 Wiley-Liss, Inc.

  12. Oral direct thrombin inhibitors or oral factor Xa inhibitors for the treatment of pulmonary embolism.

    PubMed

    Robertson, Lindsay; Kesteven, Patrick; McCaslin, James E

    2015-12-04

    Pulmonary embolism is a potentially life-threatening condition in which a clot can travel from the deep veins, most commonly in the leg, up to the lungs. Previously, a pulmonary embolism was treated with the anticoagulants heparin and vitamin K antagonists. Recently, however, two forms of direct oral anticoagulants (DOACs) have been developed: oral direct thrombin inhibitors (DTI) and oral factor Xa inhibitors. The new drugs have characteristics that may be favourable over conventional treatment, including oral administration, a predictable effect, lack of frequent monitoring or re-dosing and few known drug interactions. To date, no Cochrane review has measured the effectiveness and safety of these drugs in the long-term treatment (minimum duration of three months) of pulmonary embolism. To assess the effectiveness of oral DTIs and oral factor Xa inhibitors for the long-term treatment of pulmonary embolism. The Cochrane Vascular Trials Search Co-ordinator searched the Specialised Register (last searched January 2015) and the Cochrane Register of Studies (last searched January 2015). Clinical trials databases were also searched for details of ongoing or unpublished studies. We searched the reference lists of relevant articles retrieved by electronic searches for additional citations. We included randomised controlled trials in which patients with a pulmonary embolism confirmed by standard imaging techniques were allocated to receive an oral DTI or an oral factor Xa inhibitor for the long-term (minimum duration three months) treatment of pulmonary embolism. Two review authors (LR, JM) independently extracted the data and assessed the risk of bias in the trials. Any disagreements were resolved by discussion with the third author (PK). We used meta-analyses when we considered heterogeneity low. The two primary outcomes were recurrent venous thromboembolism and pulmonary embolism. Other outcomes included all-cause mortality and major bleeding. We calculated all outcomes

  13. Inhibitors in haemophilia A: a perspective on clotting factor products as a potential contributing factor.

    PubMed

    Mathew, P; Dinter, H; Church, N; Humphries, T J; Kulkarni, R

    2016-05-01

    The occurrence of a neutralizing antibody in previously untreated patients (PUPs) with haemophilia A appears to be the result of an intricate interplay of both genetic and environmental factors. Recently, the type of factor VIII (FVIII) product used in the PUPs population has been implicated as a risk factor for inhibitor development. The aim of this review was to explore in a systematic manner potential hypotheses for the product-related findings in these studies (i.e. differences in the expression system of the cell lines used to produce recombinant FVIII [rFVIII], differences in the administered antigen load or changes in clinical practice over time). Review of the available clinical studies illustrates the high degree of variability for the risk of inhibitor development for the same products across different studies. Differences in cell lines or antigen load were not found to provide a reasonable explanation. The possibility of changes in clinical practice over time and patient selection bias (i.e. the preferential use of one product over another in patients at higher risk for inhibitors) offers a potential explanation and should be carefully considered when evaluating the studies. © 2016 The Authors. Haemophilia Published by John Wiley & Sons Ltd.

  14. Influence of vineyard altitude on Glera grape ripening (Vitis vinifera L.): effects on aroma evolution and wine sensory profile.

    PubMed

    Alessandrini, Massimiliano; Gaiotti, Federica; Belfiore, Nicola; Matarese, Fabiola; D'Onofrio, Claudio; Tomasi, Diego

    2017-07-01

    Environmental factors have been acknowledged to greatly influence grape and wine aromas. Among them, the effect of altitude on grape aroma compounds has scarcely been debated in literature available to date. In the present study, we investigated the influence of altitude on grape composition and aroma evolution during ripening of Vitis vinifera L. cultivar Glera grown in Conegliano-Valdobbiadene DOCG area (Italy). The site at highest altitude (380 m above sea level) was warmer than the lowest site (200 m above sea level) and, even with differences in temperature in the range 1.5-2 °C, the impact of the cultivation site on grape ripening and aroma accumulation and preservation was significant. The lowest site demonstrated slower grape ripening, and grapes at harvest accumulated lower amounts of all of the main classes of aroma compounds typical of the Glera variety. Wines produced from the highest site were preferred in tasting trials for their more patent floral notes and elegance. Altitude strongly influences grape ripening evolution and flavour accumulation in the Glera grape, and this result accounts for the different styles in the sparkling wines subsequently produced. Moreover, the present study shows that aroma compound biosynthesis, particularly that of benzenoides, starts before véraison in Glera. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  15. A mechanistic modelling approach to understand 1-MCP inhibition of ethylene action and quality changes during ripening of apples.

    PubMed

    Gwanpua, Sunny George; Verlinden, Bert E; Hertog, Maarten Latm; Nicolai, Bart M; Geeraerd, Annemie H

    2017-08-01

    1-Methylcyclopropene (1-MCP) inhibits ripening in climacteric fruit by blocking ethylene receptors, preventing ethylene from binding and eliciting its action. The objective of the current study was to use mathematical models to describe 1-MCP inhibition of apple fruit ripening, and to provide a tool for predicting ethylene production, and two important quality indicators of apple fruit, firmness and background colour. A model consisting of coupled differential equations describing 1-MCP inhibition of apple ripening was developed. Data on ethylene production, expression of ethylene receptors, firmness, and background colour during ripening of untreated and 1-MCP treated apples were used to calibrate the model. An overall adjusted R 2 of 95% was obtained. The impact of time from harvest to treatment, and harvest maturity on 1-MCP efficacy was modelled. Different hypotheses on the partial response of 'Jonagold' apple to 1-MCP treatment were tested using the model. The model was validated using an independent dataset. Low 1-MCP blocking efficacy was shown to be the most likely cause of partial response for delayed 1-MCP treatment, and 1-MCP treatment of late-picked apples. Time from harvest to treatment was a more important factor than maturity for 1-MCP efficacy in 'Jonagold' apples. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  16. Sensory quality of Camembert-type cheese: Relationship between starter cultures and ripening molds.

    PubMed

    Galli, Bruno Domingues; Martin, José Guilherme Prado; da Silva, Paula Porrelli Moreira; Porto, Ernani; Spoto, Marta Helena Fillet

    2016-10-03

    Starter cultures and ripening molds used in the manufacture of moldy cheese aimed at obtaining characteristic flavors and textures considerably differ among dairy industries. Thus, the study of variables inherent to the process and their influence on sensory patterns in cheese can improve the standardization and control of the production process. The aim of this work was to study the influence of three different variables on the sensory quality of Camembert-type cheese: type of lactic bacteria, type of ripener molds and inoculation method. Batches of Camembert-type cheese were produced using O or DL-type mesophilic starter culture, ripened with Penicillium camemberti or Penicillium candidum and mold inoculation was made directly into the milk or by spraying. All batches were sensorially evaluated using Quantitative Descriptive Analysis (QDA) with panelists trained for various attributes. Among the combinations analyzed, those resulting in more typical Camembert-type cheese were those using O-type mesophilic starter culture and P. candidum maturation mold directly applied into the milk or sprayed and those using DL-type mesophilic starter and P. camemberti ripener mold applied by surface spraying. These results demonstrate, therefore, that the combination of different ripener molds, inoculation methods and starter cultures directly influences the sensory quality of Camembert-type cheese, modifying significantly its texture, appearance, aroma and taste. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Prediction of textural attributes using color values of banana (Musa sapientum) during ripening.

    PubMed

    Jaiswal, Pranita; Jha, Shyam Narayan; Kaur, Poonam Preet; Bhardwaj, Rishi; Singh, Ashish Kumar; Wadhawan, Vishakha

    2014-06-01

    Banana is an important sub-tropical fruit in international trade. It undergoes significant textural and color transformations during ripening process, which in turn influence the eating quality of the fruit. In present study, color ('L', 'a' and 'b' value) and textural attributes of bananas (peel, fruit and pulp firmness; pulp toughness; stickiness) were studied simultaneously using Hunter Color Lab and Texture Analyser, respectively, during ripening period of 10 days at ambient atmosphere. There was significant effect of ripening period on all the considered textural characteristics and color properties of bananas except color value 'b'. In general, textural descriptors (peel, fruit and pulp firmness; and pulp toughness) decreased during ripening except stickiness, while color values viz 'a' and 'b' increased with ripening barring 'L' value. Among various textural attributes, peel toughness and pulp firmness showed highest correlation (r) with 'a' value of banana peel. In order to predict textural properties using color values of banana, five types of equations (linear/polynomial/exponential/logarithmic/power) were fitted. Among them, polynomial equation was found to be the best fit (highest coefficient of determination, R(2)) for prediction of texture using color properties for bananas. The pulp firmness, peel toughness and pulp toughness showed R(2) above 0.84 with indicating its potentiality of the fitted equations for prediction of textural profile of bananas non-destructively using 'a' value.

  18. Transcriptome changes during fruit development and ripening of sweet orange (Citrus sinensis).

    PubMed

    Yu, Keqin; Xu, Qiang; Da, Xinlei; Guo, Fei; Ding, Yuduan; Deng, Xiuxin

    2012-01-10

    The transcriptome of the fruit pulp of the sweet orange variety Anliu (WT) and that of its red fleshed mutant Hong Anliu (MT) were compared to understand the dynamics and differential expression of genes expressed during fruit development and ripening. The transcriptomes of WT and MT were sampled at four developmental stages using an Illumina sequencing platform. A total of 19,440 and 18,829 genes were detected in MT and WT, respectively. Hierarchical clustering analysis revealed 24 expression patterns for the set of all genes detected, of which 20 were in common between MT and WT. Over 89% of the genes showed differential expression during fruit development and ripening in the WT. Functional categorization of the differentially expressed genes revealed that cell wall biosynthesis, carbohydrate and citric acid metabolism, carotenoid metabolism, and the response to stress were the most differentially regulated processes occurring during fruit development and ripening. A description of the transcriptomic changes occurring during fruit development and ripening was obtained in sweet orange, along with a dynamic view of the gene expression differences between the wild type and a red fleshed mutant. © 2012 Yu et al; licensee BioMed Central Ltd.

  19. Modeling the growth of Listeria monocytogenes on the surface of smear- or mold-ripened cheese.

    PubMed

    Schvartzman, M Sol; Gonzalez-Barron, Ursula; Butler, Francis; Jordan, Kieran

    2014-01-01

    Surface-ripened cheeses are matured by means of manual or mechanical technologies posing a risk of cross-contamination, if any cheeses are contaminated with Listeria monocytogenes. In predictive microbiology, primary models are used to describe microbial responses, such as growth rate over time and secondary models explain how those responses change with environmental factors. In this way, primary models were used to assess the growth rate of L. monocytogenes during ripening of the cheeses and the secondary models to test how much the growth rate was affected by either the pH and/or the water activity (aw) of the cheeses. The two models combined can be used to predict outcomes. The purpose of these experiments was to test three primary (the modified Gompertz equation, the Baranyi and Roberts model, and the Logistic model) and three secondary (the Cardinal model, the Ratowski model, and the Presser model) mathematical models in order to define which combination of models would best predict the growth of L. monocytogenes on the surface of artificially contaminated surface-ripened cheeses. Growth on the surface of the cheese was assessed and modeled. The primary models were firstly fitted to the data and the effects of pH and aw on the growth rate (μmax) were incorporated and assessed one by one with the secondary models. The Logistic primary model by itself did not show a better fit of the data among the other primary models tested, but the inclusion of the Cardinal secondary model improved the final fit. The aw was not related to the growth of Listeria. This study suggests that surface-ripened cheese should be separately regulated within EU microbiological food legislation and results expressed as counts per surface area rather than per gram.

  20. Role for the banana AGAMOUS-like gene MaMADS7 in regulation of fruit ripening and quality.

    PubMed

    Liu, Juhua; Liu, Lin; Li, Yujia; Jia, Caihong; Zhang, Jianbin; Miao, Hongxia; Hu, Wei; Wang, Zhuo; Xu, Biyu; Jin, Zhiqiang

    2015-11-01

    MADS-box transcription factors play important roles in organ development. In plants, most studies on MADS-box genes have mainly focused on flower development and only a few concerned fruit development and ripening. A new MADS-box gene named MaMADS7 was isolated from banana fruit by rapid amplification of cDNA ends (RACE) based on a MADS-box fragment obtained from a banana suppression subtractive hybridization (SSH) cDNA library. MaMADS7 is an AGAMOUS-like MADS-box gene that is preferentially expressed in the ovaries and fruits and in tobacco its protein product localizes to the nucleus. This study found that MaMADS7 expression can be induced by exogenous ethylene. Ectopic expression of MaMADS7 in tomato resulted in broad ripening phenotypes. The expression levels of seven ripening and quality-related genes, ACO1, ACS2, E4, E8, PG, CNR and PSY1 in MaMADS7 transgenic tomato fruits were greatly increased while the expression of the AG-like MADS-box gene TAGL1 was suppressed. Compared with the control, the contents of β-carotene, lycopene, ascorbic acid and organic acid in transformed tomato fruits were increased, while the contents of glucose and fructose were slightly decreased. MaMADS7 interacted with banana 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase gene 1 (MaACO1) and tomato phytoene synthase gene (LePSY1) promoters. Our results indicated that MaMADS7 plays an important role in initiating endogenous ethylene biosynthesis and fruit ripening. © 2015 Scandinavian Plant Physiology Society.

  1. Herpes Simplex Encephalitis during Treatment with Tumor Necrosis FactorInhibitors

    PubMed Central

    Bradford, Russell D.; Pettit, April C.; Wright, Patty W.; Mulligan, Mark J.; Moreland, Larry W.; McLain, David A.; Gnann, John W.; Bloch, Karen C.

    2012-01-01

    We report 3 cases of herpes simplex virus encephalitis in patients receiving tumor necrosis factor-alpha (TNF-α) inhibitors for rheumatologic disorders. Although TNF-α inhibitors have been reported to increase the risk of other infectious diseases, to our knowledge, an association between anti–TNF-α drugs and herpes simplex virus encephalitis has not been previously described. PMID:19681709

  2. Metagenomic and metatranscriptomic analysis of the microbial community in Swiss-type Maasdam cheese during ripening.

    PubMed

    Duru, Ilhan Cem; Laine, Pia; Andreevskaya, Margarita; Paulin, Lars; Kananen, Soila; Tynkkynen, Soile; Auvinen, Petri; Smolander, Olli-Pekka

    2018-05-19

    In Swiss-type cheeses, characteristic nut-like and sweet flavor develops during the cheese ripening due to the metabolic activities of cheese microbiota. Temperature changes during warm and cold room ripening, and duration of ripening can significantly change the gene expression of the cheese microbiota, which can affect the flavor formation. In this study, a metagenomic and metatranscriptomic analysis of Swiss-type Maasdam cheese was performed on samples obtained during ripening in the warm and cold rooms. We reconstructed four different bacterial genomes (Lactococcus lactis, Lactobacillus rhamnosus, Lactobacillus helveticus, and Propionibacterium freudenreichii subsp. shermanii strain JS) from the Maasdam cheese to near completeness. Based on the DNA and RNA mean coverage, Lc. lactis strongly dominated (~80-90%) within the cheese microbial community. Genome annotation showed the potential for the presence of several flavor forming pathways in these species, such as production of methanethiol, free fatty acids, acetoin, diacetyl, acetate, ethanol, and propionate. Using the metatranscriptomic data, we showed that, with the exception of Lc. lactis, the central metabolism of the microbiota was downregulated during cold room ripening suggesting that fewer flavor compounds such as acetoin and propionate were produced. In contrast, Lc. lactis genes related to the central metabolism, including the vitamin biosynthesis and homolactic fermentation, were upregulated during cold room ripening. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

  3. Use of homologous and heterologous gene expression profiling tools to characterize transcription dynamics during apple fruit maturation and ripening.

    PubMed

    Costa, Fabrizio; Alba, Rob; Schouten, Henk; Soglio, Valeria; Gianfranceschi, Luca; Serra, Sara; Musacchi, Stefano; Sansavini, Silviero; Costa, Guglielmo; Fei, Zhangjun; Giovannoni, James

    2010-10-25

    Fruit development, maturation and ripening consists of a complex series of biochemical and physiological changes that in climacteric fruits, including apple and tomato, are coordinated by the gaseous hormone ethylene. These changes lead to final fruit quality and understanding of the functional machinery underlying these processes is of both biological and practical importance. To date many reports have been made on the analysis of gene expression in apple. In this study we focused our investigation on the role of ethylene during apple maturation, specifically comparing transcriptomics of normal ripening with changes resulting from application of the hormone receptor competitor 1-methylcyclopropene. To gain insight into the molecular process regulating ripening in apple, and to compare to tomato (model species for ripening studies), we utilized both homologous and heterologous (tomato) microarray to profile transcriptome dynamics of genes involved in fruit development and ripening, emphasizing those which are ethylene regulated.The use of both types of microarrays facilitated transcriptome comparison between apple and tomato (for the later using data previously published and available at the TED: tomato expression database) and highlighted genes conserved during ripening of both species, which in turn represent a foundation for further comparative genomic studies. The cross-species analysis had the secondary aim of examining the efficiency of heterologous (specifically tomato) microarray hybridization for candidate gene identification as related to the ripening process. The resulting transcriptomics data revealed coordinated gene expression during fruit ripening of a subset of ripening-related and ethylene responsive genes, further facilitating the analysis of ethylene response during fruit maturation and ripening. Our combined strategy based on microarray hybridization enabled transcriptome characterization during normal climacteric apple ripening, as well as

  4. Use of homologous and heterologous gene expression profiling tools to characterize transcription dynamics during apple fruit maturation and ripening

    PubMed Central

    2010-01-01

    Background Fruit development, maturation and ripening consists of a complex series of biochemical and physiological changes that in climacteric fruits, including apple and tomato, are coordinated by the gaseous hormone ethylene. These changes lead to final fruit quality and understanding of the functional machinery underlying these processes is of both biological and practical importance. To date many reports have been made on the analysis of gene expression in apple. In this study we focused our investigation on the role of ethylene during apple maturation, specifically comparing transcriptomics of normal ripening with changes resulting from application of the hormone receptor competitor 1-Methylcyclopropene. Results To gain insight into the molecular process regulating ripening in apple, and to compare to tomato (model species for ripening studies), we utilized both homologous and heterologous (tomato) microarray to profile transcriptome dynamics of genes involved in fruit development and ripening, emphasizing those which are ethylene regulated. The use of both types of microarrays facilitated transcriptome comparison between apple and tomato (for the later using data previously published and available at the TED: tomato expression database) and highlighted genes conserved during ripening of both species, which in turn represent a foundation for further comparative genomic studies. The cross-species analysis had the secondary aim of examining the efficiency of heterologous (specifically tomato) microarray hybridization for candidate gene identification as related to the ripening process. The resulting transcriptomics data revealed coordinated gene expression during fruit ripening of a subset of ripening-related and ethylene responsive genes, further facilitating the analysis of ethylene response during fruit maturation and ripening. Conclusion Our combined strategy based on microarray hybridization enabled transcriptome characterization during normal climacteric

  5. Co-ordination of early and late ripening events in apples is regulated through differential sensitivities to ethylene

    PubMed Central

    Johnston, Jason W.; Gunaseelan, Kularajathaven; Pidakala, Paul; Wang, Mindy; Schaffer, Robert J.

    2009-01-01

    In this study, it is shown that anti-sense suppression of Malus domestica 1-AMINO-CYCLOPROPANE-CARBOXYLASE OXIDASE (MdACO1) resulted in fruit with an ethylene production sufficiently low to be able to assess ripening in the absence of ethylene. Exposure of these fruit to different concentrations of exogenous ethylene showed that flesh softening, volatile biosynthesis, and starch degradation, had differing ethylene sensitivity and dependency. Early ripening events such as the conversion of starch to sugars showed a low dependency for ethylene, but a high sensitivity to low concentrations of ethylene (0.01 μl l−1). By contrast, later ripening events such as flesh softening and ester volatile production showed a high dependency for ethylene but were less sensitive to low concentrations (needing 0.1 μl l−1 for a response). A sustained exposure to ethylene was required to maintain ripening, indicating that the role of ethylene may go beyond that of ripening initiation. These results suggest a conceptual model for the control of individual ripening characters in apple, based on both ethylene dependency and sensitivity. PMID:19429839

  6. Implication of Abscisic Acid on Ripening and Quality in Sweet Cherries: Differential Effects during Pre- and Post-harvest

    PubMed Central

    Tijero, Verónica; Teribia, Natalia; Muñoz, Paula; Munné-Bosch, Sergi

    2016-01-01

    Sweet cherry, a non-climacteric fruit, is usually cold-stored during post-harvest to prevent over-ripening. The aim of the study was to evaluate the role of abscisic acid (ABA) on fruit growth and ripening of this fruit, considering as well its putative implication in over-ripening and effects on quality. We measured the endogenous concentrations of ABA during the ripening of sweet cherries (Prunus avium L. var. Prime Giant) collected from orchard trees and in cherries exposed to 4°C and 23°C during 10 days of post-harvest. Furthermore, we examined to what extent endogenous ABA concentrations were related to quality parameters, such as fruit biomass, anthocyanin accumulation and levels of vitamins C and E. Endogenous concentrations of ABA in fruits increased progressively during fruit growth and ripening on the tree, to decrease later during post-harvest at 23°C. Cold treatment, however, increased ABA levels and led to an inhibition of over-ripening. Furthermore, ABA levels positively correlated with anthocyanin and vitamin E levels during pre-harvest, but not during post-harvest. We conclude that ABA plays a major role in sweet cherry development, stimulating its ripening process and positively influencing quality parameters during pre-harvest. The possible influence of ABA preventing over-ripening in cold-stored sweet cherries is also discussed. PMID:27200070

  7. Studies on physico-chemical changes during artificial ripening of banana (Musa sp) variety 'Robusta'.

    PubMed

    Kulkarni, Shyamrao Gururao; Kudachikar, V B; Keshava Prakash, M N

    2011-12-01

    Banana (Musa sp var 'Robusta') fruits harvested at 75-80% maturity were dip treated with different concentrations of ethrel (250-1,000 ppm) solution for 5 min. Ethrel at 500 ppm induced uniform ripening without impairing taste and flavour of banana. Untreated control banana fruits remained shriveled, green and failed to ripen evenly even after 8 days of storage. Fruits treated with 500 ppm of ethrel ripened well in 6 days at 20 ± 1 °C. Changes in total soluble solids, acidity, total sugars and total carotenoids showed increasing trends up to 6 days during ripening whereas fruit shear force values, pulp pH and total chlorophyll in peel showed decreasing trends. Sensory quality of ethrel treated banana fruits (fully ripe) were excellent with respect to external colour, taste, flavour and overall quality.

  8. The effect of ripening stages on the antioxidant potential of melon (Cucumis melo L.) cultivar Hikapel

    NASA Astrophysics Data System (ADS)

    Wulandari, Puji; Daryono, Budi Setiadi; Supriyadi

    2017-06-01

    Melon (Cucumis melo L.) cultivar Hikapel, a new cultivar of melon, is one of non-netted orange-fleshed melon. Non-netted orange-fleshed melon is known as source of several phytochemicals such as phenolics, flavonoids, ascorbic acid, and carotenoids. During the ripening stages there are chemical changes of the fruit including antioxidant properties. The aims of this research were to study the changes of antioxidant activity and antioxidant compound during ripening stages of melon cv. Hikapel. Melon with three ripening stages (27 DAA, 29 DAA, and 32 DAA) were harvested and analyzed their antioxidant activity, ascorbic acid, total-phenolic, -flavonoid, and -carotenoid content. The results showed that ascorbic acid and carotenoid content increased during ripening stages, whereas total phenolic and antioxidant activity decreased. The ripening stages affected antioxidant activity of Cucumis melo L. cv. Hikapel. Antioxidant activity positively correlated with ascorbic acid, total-phenolic, and -flavonoid content. On the other hand, total carotenoid negatively correlated with antioxidant activity.

  9. Development of Metal-Organic Framework for Gaseous Plant Hormone Encapsulation To Manage Ripening of Climacteric Produce.

    PubMed

    Zhang, Boce; Luo, Yaguang; Kanyuck, Kelsey; Bauchan, Gary; Mowery, Joseph; Zavalij, Peter

    2016-06-29

    Controlled ripening of climacteric fruits, such as bananas and avocados, is a critical step to provide consumers with high-quality products while reducing postharvest losses. Prior to ripening, these fruits can be stored for an extended period of time but are usually not suitable for consumption. However, once ripening is initiated, they undergo irreversible changes that lead to rapid quality loss and decay if not consumed within a short window of time. Therefore, technologies to slow the ripening process after its onset or to stimulate ripening immediately before consumption are in high demand. In this study, we developed a solid porous metal-organic framework (MOF) to encapsulate gaseous ethylene for subsequent release. We evaluated the feasibility of this technology for on-demand stimulated ripening of bananas and avocados. Copper terephthalate (CuTPA) MOF was synthesized via a solvothermal method and loaded with ethylene gas. Its crystalline structure and chemical composition were characterized by X-ray diffraction crystallography, porosity by N2 and ethylene isotherms, and morphology by electron microscopy. The MOF loaded with ethylene (MOF-ethylene) was placed inside sealed containers with preclimacteric bananas and avocados and stored at 16 °C. The headspace gas composition and fruit color and texture were monitored periodically. Results showed that this CuTPA MOF is highly porous, with a total pore volume of 0.39 cm(3)/g. A 50 mg portion of MOF-ethylene can absorb and release up to 654 μL/L of ethylene in a 4 L container. MOF-ethylene significantly accelerated the ripening-related color and firmness changes of treated bananas and avocados. This result suggests that MOF-ethylene technology could be used for postharvest application to stimulate ripening just before the point of consumption.

  10. Characterization of a calcium/calmodulin-regulated SR/CAMTA gene family during tomato fruit development and ripening

    PubMed Central

    2012-01-01

    Background Fruit ripening is a complicated development process affected by a variety of external and internal cues. It is well established that calcium treatment delays fruit ripening and senescence. However, the underlying molecular mechanisms remain unclear. Results Previous studies have shown that calcium/calmodulin-regulated SR/CAMTAs are important for modulation of disease resistance, cold sensitivity and wounding response in vegetative tissues. To study the possible roles of this gene family in fruit development and ripening, we cloned seven SR/CAMTAs, designated as SlSRs, from tomato, a model fruit-bearing crop. All seven genes encode polypeptides with a conserved DNA-binding domain and a calmodulin-binding site. Calmodulin specifically binds to the putative targeting site in a calcium-dependent manner. All SlSRs were highly yet differentially expressed during fruit development and ripening. Most notably, the expression of SlSR2 was scarcely detected at the mature green and breaker stages, two critical stages of fruit development and ripening; and SlSR3L and SlSR4 were expressed exclusively in fruit tissues. During the developmental span from 10 to 50 days post anthesis, the expression profiles of all seven SlSRs were dramatically altered in ripening mutant rin compared with wildtype fruit. By contrast, only minor alterations were noted for ripening mutant nor and Nr fruit. In addition, ethylene treatment of mature green wildtype fruit transiently stimulated expression of all SlSRs within one to two hours. Conclusions This study indicates that SlSR expression is influenced by both the Rin-mediated developmental network and ethylene signaling. The results suggest that calcium signaling is involved in the regulation of fruit development and ripening through calcium/calmodulin/SlSR interactions. PMID:22330838

  11. Sugar demand of ripening grape berries leads to recycling of surplus phloem water via the xylem.

    PubMed

    Keller, Markus; Zhang, Yun; Shrestha, Pradeep M; Biondi, Marco; Bondada, Bhaskar R

    2015-06-01

    We tested the common assumption that fleshy fruits become dependent on phloem water supply because xylem inflow declines at the onset of ripening. Using two distinct grape genotypes exposed to drought stress, we found that a sink-driven rise in phloem inflow at the beginning of ripening was sufficient to reverse drought-induced berry shrinkage. Rewatering accelerated berry growth and sugar accumulation concurrently with leaf photosynthetic recovery. Interrupting phloem flow through the peduncle prevented the increase in berry growth after rewatering, but interrupting xylem flow did not. Nevertheless, xylem flow in ripening berries, but not berry size, remained responsive to root or shoot pressurization. A mass balance analysis on ripening berries sampled in the field suggested that phloem water inflow may exceed growth and transpiration water demands. Collecting apoplastic sap from ripening berries showed that osmotic pressure increased at distinct rates in berry vacuoles and apoplast. Our results indicate that the decrease in xylem inflow at the onset of ripening may be a consequence of the sink-driven increase in phloem inflow. We propose a conceptual model in which surplus phloem water bypasses the fruit cells and partly evaporates from the berry surface and partly moves apoplastically to the xylem for outflow. © 2014 John Wiley & Sons Ltd.

  12. Mycotoxin production capability of Penicillium roqueforti in strains isolated from mould-ripened traditional Turkish civil cheese.

    PubMed

    Cakmakci, Songul; Gurses, Mustafa; Hayaloglu, A Adnan; Cetin, Bulent; Sekerci, Pinar; Dagdemir, Elif

    2015-01-01

    Mould-ripened civil is a traditional cheese produced mainly in eastern Turkey. The cheese is produced with a mixture of civil and whey curd cheeses (lor). This mixture is pressed into goat skins or plastic bags and is ripened for more than three months. Naturally occurring moulds grow on the surface and inside of the cheese during ripening. In this research, 140 Penicillium roqueforti strains were isolated from 41 samples of mould-ripened civil cheese collected from Erzurum and around towns in eastern Turkey. All strains were capable of mycotoxin production and were analysed using an HPLC method. It was established that all the strains (albeit at very low levels) produced roquefortine C, penicillic acid, mycophenolic acid and patulin. The amounts of toxins were in the ranges 0.4-47.0, 0.2-43.6, 0.1-23.1 and 0.1-2.3 mg kg(-1), respectively. Patulin levels of the samples were lower than the others. The lowest level and highest total mycotoxin levels were determined as 1.2 and 70.1 mg kg(-1) respectively. The results of this preliminary study may help in the choice of secondary cultures for mould-ripened civil cheese and other mould-ripened cheeses.

  13. Sulfated Pentagalloylglucoside is a Potent, Allosteric, and Selective Inhibitor of Factor XIa

    PubMed Central

    Al-Horani, Rami A.; Ponnusamy, Pooja; Mehta, Akul Y.; Gailani, David; Desai, Umesh R.

    2013-01-01

    Inhibition of factor XIa (FXIa) is a novel paradigm for developing anticoagulants without major bleeding consequences. We present the discovery of sulfated pentagalloylglucoside (6) as a highly selective inhibitor of human FXIa. Biochemical screening of a focused library led to the identification of 6, a sulfated aromatic mimetic of heparin. Inhibitor 6 displayed a potency of 551 nM against FXIa, which was at least 200-fold more selective than other relevant enzymes. It also prevented activation of factor IX and prolonged human plasma and whole blood clotting. Inhibitor 6 reduced VMAX of FXIa hydrolysis of chromogenic substrate without affecting the KM suggesting an allosteric mechanism. Competitive studies showed that 6 bound in the heparin-binding site of FXIa. No allosteric small molecule has been discovered to date that exhibits equivalent potency against FXIa. Inhibitor 6 is expected to open up a major route to allosteric FXIa anticoagulants with clinical relevance. PMID:23316863

  14. After-Ripening Induced Transcriptional Changes of Hormonal Genes in Wheat Seeds: The Cases of Brassinosteroids, Ethylene, Cytokinin and Salicylic Acid

    PubMed Central

    Yao, Zhen; Jordan, Mark C.; Park, Seokhoon; Ayele, Belay T.

    2014-01-01

    Maintenance and release of seed dormancy is regulated by plant hormones; their levels and seed sensitivity being the critical factors. This study reports transcriptional regulation of brassinosteroids (BR), ethylene (ET), cytokinin (CK) and salicylic acid (SA) related wheat genes by after-ripening, a period of dry storage that decays dormancy. Changes in the expression of hormonal genes due to seed after-ripening did not occur in the anhydrobiotic state but rather in the hydrated state. After-ripening induced dormancy decay appears to be associated with imbibition mediated increase in the synthesis and signalling of BR, via transcriptional activation of de-etiolated2, dwarf4 and brassinosteroid signaling kinase, and repression of brassinosteroid insensitive 2. Our analysis is also suggestive of the significance of increased ET production, as reflected by enhanced transcription of 1-aminocyclopropane-1-carboxylic acid oxidase in after-ripened seeds, and tight regulation of seed response to ET in regulating dormancy decay. Differential transcriptions of lonely guy, zeatin O-glucosyltransferases and cytokinin oxidases, and pseudo-response regulator between dormant and after-ripened seeds implicate CK in the regulation of seed dormancy in wheat. Our analysis also reflects the association of dormancy decay in wheat with seed SA level and NPR independent SA signaling that appear to be regulated transcriptionally by phenylalanine ammonia lyase, and whirly and suppressor of npr1 inducible1 genes, respectively. Co-expression clustering of the hormonal genes implies the significance of synergistic and antagonistic interaction between the different plant hormones in regulating wheat seed dormancy. These results contribute to further our understanding of the molecular features controlling seed dormancy in wheat. PMID:24498132

  15. Characterization of an AGAMOUS-like MADS Box Protein, a Probable Constituent of Flowering and Fruit Ripening Regulatory System in Banana

    PubMed Central

    Roy Choudhury, Swarup; Roy, Sujit; Nag, Anish; Singh, Sanjay Kumar; Sengupta, Dibyendu N.

    2012-01-01

    The MADS-box family of genes has been shown to play a significant role in the development of reproductive organs, including dry and fleshy fruits. In this study, the molecular properties of an AGAMOUS like MADS box transcription factor in banana cultivar Giant governor (Musa sp, AAA group, subgroup Cavendish) has been elucidated. We have detected a CArG-box sequence binding AGAMOUS MADS-box protein in banana flower and fruit nuclear extracts in DNA-protein interaction assays. The protein fraction in the DNA-protein complex was analyzed by mass spectrometry and using this information we have obtained the full length cDNA of the corresponding protein. The deduced protein sequence showed ∼95% amino acid sequence homology with MA-MADS5, a MADS-box protein described previously from banana. We have characterized the domains of the identified AGAMOUS MADS-box protein involved in DNA binding and homodimer formation in vitro using full-length and truncated versions of affinity purified recombinant proteins. Furthermore, in order to gain insight about how DNA bending is achieved by this MADS-box factor, we performed circular permutation and phasing analysis using the wild type recombinant protein. The AGAMOUS MADS-box protein identified in this study has been found to predominantly accumulate in the climacteric fruit pulp and also in female flower ovary. In vivo and in vitro assays have revealed specific binding of the identified AGAMOUS MADS-box protein to CArG-box sequence in the promoters of major ripening genes in banana fruit. Overall, the expression patterns of this MADS-box protein in banana female flower ovary and during various phases of fruit ripening along with the interaction of the protein to the CArG-box sequence in the promoters of major ripening genes lead to interesting assumption about the possible involvement of this AGAMOUS MADS-box factor in banana fruit ripening and floral reproductive organ development. PMID:22984496

  16. Characterization of an AGAMOUS-like MADS box protein, a probable constituent of flowering and fruit ripening regulatory system in banana.

    PubMed

    Roy Choudhury, Swarup; Roy, Sujit; Nag, Anish; Singh, Sanjay Kumar; Sengupta, Dibyendu N

    2012-01-01

    The MADS-box family of genes has been shown to play a significant role in the development of reproductive organs, including dry and fleshy fruits. In this study, the molecular properties of an AGAMOUS like MADS box transcription factor in banana cultivar Giant governor (Musa sp, AAA group, subgroup Cavendish) has been elucidated. We have detected a CArG-box sequence binding AGAMOUS MADS-box protein in banana flower and fruit nuclear extracts in DNA-protein interaction assays. The protein fraction in the DNA-protein complex was analyzed by mass spectrometry and using this information we have obtained the full length cDNA of the corresponding protein. The deduced protein sequence showed ~95% amino acid sequence homology with MA-MADS5, a MADS-box protein described previously from banana. We have characterized the domains of the identified AGAMOUS MADS-box protein involved in DNA binding and homodimer formation in vitro using full-length and truncated versions of affinity purified recombinant proteins. Furthermore, in order to gain insight about how DNA bending is achieved by this MADS-box factor, we performed circular permutation and phasing analysis using the wild type recombinant protein. The AGAMOUS MADS-box protein identified in this study has been found to predominantly accumulate in the climacteric fruit pulp and also in female flower ovary. In vivo and in vitro assays have revealed specific binding of the identified AGAMOUS MADS-box protein to CArG-box sequence in the promoters of major ripening genes in banana fruit. Overall, the expression patterns of this MADS-box protein in banana female flower ovary and during various phases of fruit ripening along with the interaction of the protein to the CArG-box sequence in the promoters of major ripening genes lead to interesting assumption about the possible involvement of this AGAMOUS MADS-box factor in banana fruit ripening and floral reproductive organ development.

  17. Electronic eye for the prediction of parameters related to grape ripening.

    PubMed

    Orlandi, G; Calvini, R; Pigani, L; Foca, G; Vasile Simone, G; Antonelli, A; Ulrici, A

    2018-08-15

    An electronic eye (EE) for fast and easy evaluation of grape phenolic ripening has been developed. For this purpose, berries of different grape varieties were collected at different harvest times from veraison to maturity, then an amount of the derived must was deposited on a white sheet of absorbent paper to obtain a sort of paper chromatography. Thus, RGB images of the must spots were collected using a flatbed scanner and converted into one-dimensional signals, named colourgrams, which codify the colour properties of the images. The dataset of colourgrams was used to build calibration models to relate the colour of the images with the phenolic composition of the samples - determined by reference analytical methods - and therefore to follow the ripening trend. Satisfactory calibration models were obtained for the prediction of the most important parameters related to phenolic ripening of grapes, such as colour index, tonality, total anthocyanins content, malvidin-3-O-glucoside and petunidin-3-O-glucoside. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Microbiological and biochemical aspects of Camembert-type cheeses depend on atmospheric composition in the ripening chamber.

    PubMed

    Leclercq-Perlat, M-N; Picque, D; Riahi, H; Corrieu, G

    2006-08-01

    Camembert-type cheeses were prepared from pasteurized milk seeded with Kluyveromyces lactis, Geotrichum candidum, Penicillium camemberti, and Brevibacterium aurantiacum. Microorganism growth and biochemical dynamics were studied in relation to ripening chamber CO(2) atmospheric composition using 31 descriptors based on kinetic data. The chamber ripening was carried out under 5 different controlled atmospheres: continuously renewed atmosphere, periodically renewed atmosphere, no renewed atmosphere, and 2 for which CO(2) was either 2% or 6%. All microorganism dynamics depended on CO(2) level. Kluyveromyces lactis was not sensitive to CO(2) during its growth phases, but its death did depend on it. An increase of CO(2) led to a significant improvement in G. candidum. Penicillium camemberti mycelium development was enhanced by 2% CO(2). The equilibrium between P. camemberti and G. candidum populations was disrupted in favor of the yeast when CO(2) was higher than 4%. Growth of B. aurantiacum depended more on O(2) than on CO(2). Two ripening progressions were observed in relation to the presence of CO(2) at the beginning of ripening: in the presence of CO(2), the ripening was fast-slow, and in the absence of CO(2), it was slow-fast. The underrind was too runny if CO(2) was equal to or higher than 6%. The nitrogen substrate progressions were slightly related to ripening chamber CO(2) and O(2) levels. During chamber ripening, the best atmospheric condition to produce an optimum between microorganism growth, biochemical dynamics, and cheese appearance was a constant CO(2) level close to 2%.

  19. Coupling of Physiological and Proteomic Analysis to Understand the Ethylene- and Chilling-Induced Kiwifruit Ripening Syndrome

    PubMed Central

    Minas, Ioannis S.; Tanou, Georgia; Karagiannis, Evangelos; Belghazi, Maya; Molassiotis, Athanassios

    2016-01-01

    Kiwifruit [Actinidia deliciosa (A. Chev.) C.F. Liang et A.R. Ferguson, cv. “Hayward”] is classified as climacteric fruit and the initiation of endogenous ethylene production following harvest is induced by exogenous ethylene or chilling exposure. To understand the biological basis of this “dilemma,” kiwifruit ripening responses were characterized at 20°C following treatments with exogenous ethylene (100 μL L−1, 20°C, 24 h) or/and chilling temperature (0°C, 10 days). All treatments elicited kiwifruit ripening and induced softening and endogenous ethylene biosynthesis, as determined by 1-aminocyclopropane-1-carboxylic acid (ACC) content and ACC synthase (ACS) and ACC oxidase (ACO) enzyme activities after 10 days of ripening at 20°C. Comparative proteomic analysis using two-dimensional gel electrophoresis (2DE-PAGE) and nanoscale liquid chromatography coupled to tandem mass spectrometry (nanoLC-MS/MS) revealed 81 kiwifruit proteins associated with ripening. Thirty-one kiwifruit proteins were identified as commonly regulated by the three treatments accompanied by dynamic changes of 10 proteins specific to exogenous ethylene, 2 to chilling treatment, and 12 to their combination. Ethylene and/or chilling-responsive proteins were mainly involved in disease/defense, energy, protein destination/storage, and cell structure/cell wall. Interactions between the identified proteins were demonstrated by bioinformatics analysis, allowing a more complete insight into biological pathways and molecular functions affected by ripening. The present approach provides a quantitative basis for understanding the ethylene- and chilling-induced kiwifruit ripening and climacteric fruit ripening in general. PMID:26913040

  20. Simultaneous application of ethylene and 1-MCP affects banana ripening features during storage.

    PubMed

    Botondi, Rinaldo; De Sanctis, Federica; Bartoloni, Serena; Mencarelli, Fabio

    2014-08-01

    In order to avoid the ripening blocking effect of 1-MCP (1-methylcyclopropene) on bananas when applied before ethylene commercial treatment, 1-MCP in combination with 'CD ethylene' (ethylene-cyclodextrin complex) was used in gas formulations: 300 nmol mol(-1) 1-MCP + 1200, 2400 or 4800 nmol mol(-1) ethylene (ETH). Control bananas received 1-MCP alone or 4800 nmol mol(-1) ethylene alone or no treatment. Treatments were done on overseas shipped bananas, at 14 °C, 90% relative humidity (RH), for 16 h; the bananas were stored under the same atmospheric conditions. After 4 or 12 days the bananas were commercially treated with 500 µmol mol(-1) ethylene. A 300 nmol mol(-1) 1-MCP treatment significantly blocked banana ripening in terms of physiological and technological parameters, inhibiting ethylene production and respiration, despite the commercial ethylene treatment. The application of 300 nmol mol(-1) 1-MCP + 1200 or 2400 nmol mol(-1) ethylene delayed ripening but with a regular pattern. A 300 nmol mol(-1) 1-MCP + 4800 nmol mol(-1) ethylene application did not delay ripening as did 4800 nmol mol(-1) ethylene treatment. The development of black spots was closely associated with advanced ripening/senescence of fruits. The combined 300 nmol mol(-1) 1-MCP + 1200 or 2400 nmol mol(-1) ethylene treatment appears to be a promising treatment to extend banana storage, following overseas shipping. © 2014 Society of Chemical Industry.

  1. Changes in sugars, acids, and volatiles during ripening of koubo [Cereus peruvianus (L.) Miller] fruits.

    PubMed

    Ninio, Racheli; Lewinsohn, Efraim; Mizrahi, Yosef; Sitrit, Yaron

    2003-01-29

    The columnar cactus Cereus peruvianus (L.) Miller, Cactaceae (koubo), is grown commercially in Israel. The unripe fruits are green, and the color changes to violet and then to red when the fruit is fully ripe. The content of soluble sugars was found to increase 5-fold during ripening. Glucose and fructose were the main sugars accumulated in the fruit pulp, and each increased from 0.5 to 5.5 g/100 g fresh weight during ripening. The polysaccharides content decreased during ripening from 1.4 to 0.4 g/100 g fresh weight. The titratable acidity decreased and the pH increased during ripening. The major organic acid found in the fruit was malic acid, which decreased from 0.75 g/100 g fresh weight at the mature green stage to 0.355 g/100 g fresh weight in ripe fruits. Citric, succinic, and oxalic acids were found in concentrations lower than 0.07 g/100 g fresh weight. Prominent accumulation of aroma volatiles occurred toward the end of the ripening process. The main volatile found in the ripe fruit was linalool, reaching concentrations of 1.5-3.5 microg/g fresh weight.

  2. Tocopherols in rose hips (Rosa spp.) during ripening.

    PubMed

    Andersson, Staffan C; Olsson, Marie E; Gustavsson, Karl-Erik; Johansson, Eva; Rumpunen, Kimmo

    2012-08-15

    Rose hips are used as a food ingredient and in health products. They are rich in various bioactive compounds such as carotenoids and vitamin C, but data on their vitamin E content (tocopherols and tocotrienols) are limited. In this study, four different species of Rosa were analysed for tocopherol and tocotrienol content during ripening in three different years. Only α- and γ-tocopherol were found in the fleshy parts of the rose hips, and the tocopherol content and vitamin E activity varied depending on date of harvesting, species and year. The amount of vitamin E activity differed between species of Rosa and years, whereas the changes during ripening were relatively small. The choice of species must be considered if tocopherol content is to be optimised when rose hips are used as a food ingredient. Copyright © 2012 Society of Chemical Industry.

  3. Direct observation of small cluster mobility and ripening

    NASA Technical Reports Server (NTRS)

    Heinemann, K.; Poppa, H.

    1976-01-01

    Direct evidence is reported for the simultaneous occurrence of Ostwald ripening and short-distance cluster mobility during annealing of discontinuous metal films on clean amorphous substrates. The annealing characteristics of very thin particulate deposits of silver on amorphized clean surfaces of single-crystalline thin graphite substrates have been studied by in situ transmission electron microscopy (TEM) under controlled environmental conditions in the temperature range from 25 to 450 C. It was possible to monitor all stages of the experiments by TEM observation of the same specimen area. Slow Ostwald ripening was found to occur over the entire temperature range, but the overriding surface transport mechanism was short-distance cluster mobility. This was concluded from in situ observations of individual particles during annealing and from measurements of cluster size distributions, cluster number densities, area coverages, and mean cluster diameters.

  4. Segregating gas from melt: an experimental study of the Ostwald ripening of vapor bubbles in magmas

    USGS Publications Warehouse

    Lautze, Nicole C.; Sisson, Thomas W.; Mangan, Margaret T.; Grove, Timothy L.

    2011-01-01

    Diffusive coarsening (Ostwald ripening) of H2O and H2O-CO2 bubbles in rhyolite and basaltic andesite melts was studied with elevated temperature–pressure experiments to investigate the rates and time spans over which vapor bubbles may enlarge and attain sufficient buoyancy to segregate in magmatic systems. Bubble growth and segregation are also considered in terms of classical steady-state and transient (non-steady-state) ripening theory. Experimental results are consistent with diffusive coarsening as the dominant mechanism of bubble growth. Ripening is faster in experiments saturated with pure H2O than in those with a CO2-rich mixed vapor probably due to faster diffusion of H2O than CO2 through the melt. None of the experimental series followed the time1/3 increase in mean bubble radius and time-1 decrease in bubble number density predicted by classical steady-state ripening theory. Instead, products are interpreted as resulting from transient regime ripening. Application of transient regime theory suggests that bubbly magmas may require from days to 100 years to reach steady-state ripening conditions. Experimental results, as well as theory for steady-state ripening of bubbles that are immobile or undergoing buoyant ascent, indicate that diffusive coarsening efficiently eliminates micron-sized bubbles and would produce mm-sized bubbles in 102–104 years in crustal magma bodies. Once bubbles attain mm-sizes, their calculated ascent rates are sufficient that they could transit multiple kilometers over hundreds to thousands of years through mafic and silicic melt, respectively. These results show that diffusive coarsening can facilitate transfer of volatiles through, and from, magmatic systems by creating bubbles sufficiently large for rapid ascent.

  5. Antioxidant compounds of kiwifruit during post-ripening process at ambient temperature

    NASA Astrophysics Data System (ADS)

    Liang, D.; Lv, X. L.; Wang, J.; Xia, H.; Xie, Y.; Li, M. Z.; Wang, Y. Z.

    2017-09-01

    Kiwifruit is well-known for an excellent source of antioxidants. In this study, contents of total phenolics (TPC), total flavonoids (TFC), total flavanols (TFAC) and vitamin C were investigated in different fruit tissues during post-ripening process at ambient temperature. The results explored that TPC and TFC showed declining trend with the increase in storage interval in different tissues. TFAC raised with the increase in storage interval in different fruit tissues, while was followed a decrease in later process. Vitamin C content was stable in outer and inner pericarp in prometaphase of post-ripening.

  6. Comparative transcriptome analysis reveals distinct ethylene-independent regulation of ripening in response to low temperature in kiwifruit.

    PubMed

    Asiche, William O; Mitalo, Oscar W; Kasahara, Yuka; Tosa, Yasuaki; Mworia, Eric G; Owino, Willis O; Ushijima, Koichiro; Nakano, Ryohei; Yano, Kentaro; Kubo, Yasutaka

    2018-03-21

    Kiwifruit are classified as climacteric since exogenous ethylene (or its analogue propylene) induces rapid ripening accompanied by ethylene production under positive feedback regulation. However, most of the ripening-associated changes (Phase 1 ripening) in kiwifruit during storage and on-vine occur largely in the absence of any detectable ethylene. This ripening behavior is often attributed to basal levels of system I ethylene, although it is suggested to be modulated by low temperature. To elucidate the mechanisms regulating Phase 1 ripening in kiwifruit, a comparative transcriptome analysis using fruit continuously exposed to propylene (at 20 °C), and during storage at 5 °C and 20 °C was conducted. Propylene exposure induced kiwifruit softening, reduction of titratable acidity (TA), increase in soluble solids content (SSC) and ethylene production within 5 days. During storage, softening and reduction of TA occurred faster in fruit at 5 °C compared to 20 °C although no endogenous ethylene production was detected. Transcriptome analysis revealed 3761 ripening-related differentially expressed genes (DEGs), of which 2742 were up-regulated by propylene while 1058 were up-regulated by low temperature. Propylene exclusively up-regulated 2112 DEGs including those associated with ethylene biosynthesis and ripening such as AcACS1, AcACO2, AcPL1, AcXET1, Acβ-GAL, AcAAT, AcERF6 and AcNAC7. Similarly, low temperature exclusively up-regulated 467 DEGS including AcACO3, AcPL2, AcPMEi, AcADH, Acβ-AMY2, AcGA2ox2, AcNAC5 and AcbZIP2 among others. A considerable number of DEGs such as AcPG, AcEXP1, AcXET2, Acβ-AMY1, AcGA2ox1, AcNAC6, AcMADS1 and AcbZIP1 were up-regulated by either propylene or low temperature. Frequent 1-MCP treatments failed to inhibit the accelerated ripening and up-regulation of associated DEGs by low temperature indicating that the changes were independent of ethylene. On-vine kiwifruit ripening proceeded in the absence of any detectable

  7. Transcriptional regulation of anthocyanin biosynthesis in ripening fruits of grapevine under seasonal water deficit.

    PubMed

    Castellarin, Simone D; Pfeiffer, Antonella; Sivilotti, Paolo; Degan, Mirko; Peterlunger, Enrico; DI Gaspero, Gabriele

    2007-11-01

    Anthocyanin biosynthesis is strongly up-regulated in ripening fruit of grapevines (Vitis vinifera L.) grown under drought conditions. We investigated the effects of long-term water deficit on the expression of genes coding for flavonoid and anthocyanin biosynthetic enzymes and related transcription factors, genes sensitive to endogenous [sugars, abscisic acid (ABA)] and environmental (light) stimuli connected to drought stress, and genes developmentally regulated in ripening berries. Total anthocyanin content has increased at harvest in water-stressed (WS) fruits by 37-57% in two consecutive years. At least 84% of the total variation in anthocyanin content was explained by the linear relationship between the integral of mRNA accumulation of the specific anthocyanin biosynthetic gene UDP-glucose : flavonoid 3-O-glucosyltransferase (UFGT) and metabolite content during time series from véraison through ripening. Chalcone synthase (CHS2, CHS3) and flavanone 3-hydroxylase (F3H) genes of the flavonoid pathway showed high correlation as well. Genes coding for flavonoid 3',5'-hydroxylase (F3'5'H) and O-methyltransferase (OMT) were also up-regulated in berries from dehydrated plants in which anthocyanin composition enriched in more hydroxylated and more methoxylated derivatives such as malvidin and peonidin, the grape anthocyanins to which human gastric bilitranslocase displays the highest affinity. The induction in WS plants of structural and regulatory genes of the flavonoid pathway and of genes that trigger brassinosteroid hormonal onset of maturation suggested that the interrelationships between developmental and environmental signalling pathways were magnified by water deficit which actively promoted fruit maturation and, in this context, anthocyanin biosynthesis.

  8. The Jasmonate-Activated Transcription Factor MdMYC2 Regulates ETHYLENE RESPONSE FACTOR and Ethylene Biosynthetic Genes to Promote Ethylene Biosynthesis during Apple Fruit Ripening[OPEN

    PubMed Central

    Xu, Yaxiu; Zhang, Lichao; Ji, Yinglin; Tan, Dongmei; Yuan, Hui

    2017-01-01

    The plant hormone ethylene is critical for ripening in climacteric fruits, including apple (Malus domestica). Jasmonate (JA) promotes ethylene biosynthesis in apple fruit, but the underlying molecular mechanism is unclear. Here, we found that JA-induced ethylene production in apple fruit is dependent on the expression of MdACS1, an ACC synthase gene involved in ethylene biosynthesis. The expression of MdMYC2, encoding a transcription factor involved in the JA signaling pathway, was enhanced by MeJA treatment in apple fruits, and MdMYC2 directly bound to the promoters of both MdACS1 and the ACC oxidase gene MdACO1 and enhanced their transcription. Furthermore, MdMYC2 bound to the promoter of MdERF3, encoding a transcription factor involved in the ethylene-signaling pathway, thereby activating MdACS1 transcription. We also found that MdMYC2 interacted with MdERF2, a suppressor of MdERF3 and MdACS1. This protein interaction prevented MdERF2 from interacting with MdERF3 and from binding to the MdACS1 promoter, leading to increased transcription of MdACS1. Collectively, these results indicate that JA promotes ethylene biosynthesis through the regulation of MdERFs and ethylene biosynthetic genes by MdMYC2. PMID:28550149

  9. Complete genome sequence of Corynebacterium variabile DSM 44702 isolated from the surface of smear-ripened cheeses and insights into cheese ripening and flavor generation

    PubMed Central

    2011-01-01

    Background Corynebacterium variabile is part of the complex microflora on the surface of smear-ripened cheeses and contributes to the development of flavor and textural properties during cheese ripening. Still little is known about the metabolic processes and microbial interactions during the production of smear-ripened cheeses. Therefore, the gene repertoire contributing to the lifestyle of the cheese isolate C. variabile DSM 44702 was deduced from the complete genome sequence to get a better understanding of this industrial process. Results The chromosome of C. variabile DSM 44702 is composed of 3, 433, 007 bp and contains 3, 071 protein-coding regions. A comparative analysis of this gene repertoire with that of other corynebacteria detected 1, 534 predicted genes to be specific for the cheese isolate. These genes might contribute to distinct metabolic capabilities of C. variabile, as several of them are associated with metabolic functions in cheese habitats by playing roles in the utilization of alternative carbon and sulphur sources, in amino acid metabolism, and fatty acid degradation. Relevant C. variabile genes confer the capability to catabolize gluconate, lactate, propionate, taurine, and gamma-aminobutyric acid and to utilize external caseins. In addition, C. variabile is equipped with several siderophore biosynthesis gene clusters for iron acquisition and an exceptional repertoire of AraC-regulated iron uptake systems. Moreover, C. variabile can produce acetoin, butanediol, and methanethiol, which are important flavor compounds in smear-ripened cheeses. Conclusions The genome sequence of C. variabile provides detailed insights into the distinct metabolic features of this bacterium, implying a strong adaption to the iron-depleted cheese surface habitat. By combining in silico data obtained from the genome annotation with previous experimental knowledge, occasional observations on genes that are involved in the complex metabolic capacity of C. variabile

  10. Metabolomics of dates (Phoenix dactylifera) reveals a highly dynamic ripening process accounting for major variation in fruit composition.

    PubMed

    Diboun, Ilhame; Mathew, Sweety; Al-Rayyashi, Maryam; Elrayess, Mohamed; Torres, Maria; Halama, Anna; Méret, Michaël; Mohney, Robert P; Karoly, Edward D; Malek, Joel; Suhre, Karsten

    2015-12-16

    Dates are tropical fruits with appreciable nutritional value. Previous attempts at global metabolic characterization of the date metabolome were constrained by small sample size and limited geographical sampling. In this study, two independent large cohorts of mature dates exhibiting substantial diversity in origin, varieties and fruit processing conditions were measured by metabolomics techniques in order to identify major determinants of the fruit metabolome. Multivariate analysis revealed a first principal component (PC1) significantly associated with the dates' countries of production. The availability of a smaller dataset featuring immature dates from different development stages served to build a model of the ripening process in dates, which helped reveal a strong ripening signature in PC1. Analysis revealed enrichment in the dry type of dates amongst fruits with early ripening profiles at one end of PC1 as oppose to an overrepresentation of the soft type of dates with late ripening profiles at the other end of PC1. Dry dates are typical to the North African region whilst soft dates are more popular in the Gulf region, which partly explains the observed association between PC1 and geography. Analysis of the loading values, expressing metabolite correlation levels with PC1, revealed enrichment patterns of a comprehensive range of metabolite classes along PC1. Three distinct metabolic phases corresponding to known stages of date ripening were observed: An early phase enriched in regulatory hormones, amines and polyamines, energy production, tannins, sucrose and anti-oxidant activity, a second phase with on-going phenylpropanoid secondary metabolism, gene expression and phospholipid metabolism and a late phase with marked sugar dehydration activity and degradation reactions leading to increased volatile synthesis. These data indicate the importance of date ripening as a main driver of variation in the date metabolome responsible for their diverse nutritional and

  11. Effects of atmospheric composition on respiratory behavior, weight loss, and appearance of Camembert-type cheeses during chamber ripening.

    PubMed

    Picque, D; Leclercq-Perlat, M-N; Corrieu, G

    2006-08-01

    Respiratory activity, weight loss, and appearance of Camembert-type cheeses were studied during chamber ripening in relation to atmospheric composition. Cheese ripening was carried out in chambers under continuously renewed, periodically renewed, or nonrenewed gaseous atmospheres or under a CO(2) concentration kept constant at either 2 or 6% throughout the chamber-ripening process. It was found that overall atmospheric composition, and especially CO(2) concentration, of the ripening chamber affected respiratory activity. When CO(2) was maintained at either 2 or 6%, O(2) consumption and CO(2) production (and their kinetics) were higher compared with ripening trials carried out without regulating CO(2) concentration over time. Global weight loss was maximal under continuously renewed atmospheric conditions. In this case, the airflow increased exchanges between cheeses and the atmosphere. The ratio between water evaporation and CO(2) release also depended on atmospheric composition, especially CO(2) concentration. The thickening of the creamy underrind increased more quickly when CO(2) was present in the chamber from the beginning of the ripening process. However, CO(2) concentrations higher than 2% negatively influenced the appearance of the cheeses.

  12. The Jasmonate-Activated Transcription Factor MdMYC2 Regulates ETHYLENE RESPONSE FACTOR and Ethylene Biosynthetic Genes to Promote Ethylene Biosynthesis during Apple Fruit Ripening.

    PubMed

    Li, Tong; Xu, Yaxiu; Zhang, Lichao; Ji, Yinglin; Tan, Dongmei; Yuan, Hui; Wang, Aide

    2017-06-01

    The plant hormone ethylene is critical for ripening in climacteric fruits, including apple ( Malus domestica ). Jasmonate (JA) promotes ethylene biosynthesis in apple fruit, but the underlying molecular mechanism is unclear. Here, we found that JA-induced ethylene production in apple fruit is dependent on the expression of MdACS1 , an ACC synthase gene involved in ethylene biosynthesis. The expression of MdMYC2 , encoding a transcription factor involved in the JA signaling pathway, was enhanced by MeJA treatment in apple fruits, and MdMYC2 directly bound to the promoters of both MdACS1 and the ACC oxidase gene MdACO1 and enhanced their transcription. Furthermore, MdMYC2 bound to the promoter of MdERF3 , encoding a transcription factor involved in the ethylene-signaling pathway, thereby activating MdACS1 transcription. We also found that MdMYC2 interacted with MdERF2, a suppressor of MdERF3 and MdACS1 This protein interaction prevented MdERF2 from interacting with MdERF3 and from binding to the MdACS1 promoter, leading to increased transcription of MdACS1 Collectively, these results indicate that JA promotes ethylene biosynthesis through the regulation of MdERFs and ethylene biosynthetic genes by MdMYC2. © 2017 American Society of Plant Biologists. All rights reserved.

  13. Effect of different ripening stages on walnut kernel quality: antioxidant activities, lipid characterization and antibacterial properties.

    PubMed

    Amin, Furheen; Masoodi, F A; Baba, Waqas N; Khan, Asma Ashraf; Ganie, Bashir Ahmad

    2017-11-01

    Packing tissue between and around the kernel halves just turning brown (PTB) is a phenological indicator of kernel ripening at harvest in walnuts. The effect of three ripening stages (Pre-PTB, PTB and Post-PTB) on kernel quality characteristics, mineral composition, lipid characterization, sensory analysis, antioxidant and antibacterial activity were investigated in fresh kernels of indigenous numbered walnut selection of Kashmir valley "SKAU-02". Proximate composition, physical properties and sensory analysis of walnut kernels showed better results for Pre-PTB and PTB while higher mineral content was seen for kernels at Post-PTB stage in comparison to other stages of ripening. Kernels showed significantly higher levels of Omega-3 PUFA (C18:3 n3 ) and low n6/n3 ratio when harvested at Pre-PTB and PTB stages. The highest phenolic content and antioxidant activity was observed at the first stage of ripening and a steady decrease was observed at later stages. TBARS values increased as ripening advanced but did not show any significant difference in malonaldehyde formation during early ripening stages whereas it showed marked increase in walnut kernels at post-PTB stage. Walnut extracts inhibited growth of Gram-positive bacteria ( B. cereus, B. subtilis, and S. aureus ) with respective MICs of 1, 1 and 5 mg/mL and gram negative bacteria ( E. coli, P. and K. pneumonia ) with MIC of 100 mg/mL. Zone of inhibition obtained against all the bacterial strains from walnut kernel extracts increased with increase in the stage of ripening. It is concluded that Pre-PTB harvest stage with higher antioxidant activities, better fatty acid profile and consumer acceptability could be preferred harvesting stage for obtaining functionally superior walnut kernels.

  14. Ripening-Related Gene from Avocado Fruit 1

    PubMed Central

    McGarvey, Douglas J.; Sirevåg, Reidun; Christoffersen, Rolf E.

    1992-01-01

    Fruit ripening involves a series of changes in gene expression regulated by the phytohormone ethylene. AVOe3, a ripening-related gene in avocado fruit (Persea americana Mill. cv Hass), was characterized with regard to its ethylene-regulated expression. The AVOe3 mRNA and immunopositive protein were induced in mature fruit within 12 hours of propylene treatment. The AVOe3 mRNA levels reached a maximum 1 to 2 days before the ethylene climacteric, whereas the immunopositive protein continued to accumulate. RNA selected by the pAVOe3 cDNA clone encoded a polypeptide with molecular mass of 34 kilodaltons, corresponding to the molecular mass of the AVOe3 protein determined by immunoblots. The protein was soluble, remaining in solution at 100,000 gravity and eluted as a monomer on gel filtration. Because of its pattern of induction and relationship to an ethylene-related gene of tomato, the possible involvement of AVOe3 in ethylene biosynthesis is discussed. ImagesFigure 1Figure 2Figure 3Figure 4Figure 5Figure 6 PMID:16668676

  15. Spermidine affects the transcriptome responses to high temperature stress in ripening tomato fruit.

    PubMed

    Cheng, Lin; Sun, Rong-rong; Wang, Fei-yan; Peng, Zhen; Kong, Fu-ling; Wu, Jian; Cao, Jia-shu; Lu, Gang

    2012-04-01

    High temperature adversely affects quality and yield of tomato fruit. Polyamine can alleviate heat injury in plants. This study is aimed to investigate the effects of polyamine and high temperature on transcriptional profiles in ripening tomato fruit. An Affymetrix tomato microarray was used to evaluate changes in gene expression in response to exogenous spermidine (Spd, 1 mmol/L) and high temperature (33/27 °C) treatments in tomato fruits at mature green stage. Of the 10101 tomato probe sets represented on the array, 127 loci were differentially expressed in high temperature-treated fruits, compared with those under normal conditions, functionally characterized by their involvement in signal transduction, defense responses, oxidation reduction, and hormone responses. However, only 34 genes were up-regulated in Spd-treated fruits as compared with non-treated fruits, which were involved in primary metabolism, signal transduction, hormone responses, transcription factors, and stress responses. Meanwhile, 55 genes involved in energy metabolism, cell wall metabolism, and photosynthesis were down-regulated in Spd-treated fruits. Our results demonstrated that Spd might play an important role in regulation of tomato fruit response to high temperature during ripening stage.

  16. Role of sucrose phosphate synthase in sucrose biosynthesis in ripening bananas and its relationship to the respiratory climacteric.

    PubMed

    Hubbard, N L; Pharr, D M; Huber, S C

    1990-09-01

    During ripening of bananas (Musa spp. [AAA group, Cavendish subgroup]), there is a massive conversion of starch to sucrose. Also during ripening there is a rise in respiration known as the respiratory climacteric. In this study changes in carbohydrate content, activities of starch and sucrose metabolizing enzymes, and respiration were measured to assess their potential interrelationships. Sucrose phosphate synthase activity increased dramatically during the first 4 days after initiation of ripening by ethylene treatment. Starch concentration decreased and sucrose concentration increased during this time period. Developmental changes in sucrose phosphate synthase activity were measured with limiting substrate (plus Pi) and saturating substrate concentrations. Activities were not parallel under the two assay conditions, providing tentative evidence that kinetically different forms of the enzyme may exist at different stages of ripening. Sucrose accumulation rate was most highly correlated with sucrose phosphate synthase activity assayed with limiting substrate concentrations (plus Pi). The cumulative amount of CO(2) respired during ripening was positively correlated with sugar accumulation (R(2) = 0.97). From this linear regression it was calculated that a constant 0.605 millimoles of CO(2) was evolved per mole of sucrose formed throughout ripening. Using this quantity, the percentage of the total respiratory ATP produced which was required for the conversion of starch to sucrose was calculated assuming different models for carbon export from the amyloplast. The results suggest that sucrose biosynthesis during ripening constitutes a significant sink for respiratory ATP.

  17. SEPALLATA1/2-suppressed mature apples have low ethylene, high auxin and reduced transcription of ripening-related genes

    PubMed Central

    Schaffer, Robert J.; Ireland, Hilary S.; Ross, John J.; Ling, Toby J.; David, Karine M.

    2012-01-01

    Background and aims Fruit ripening is an important developmental trait in fleshy fruits, making the fruit palatable for seed dispersers. In some fruit species, there is a strong association between auxin concentrations and fruit ripening. We investigated the relationship between auxin concentrations and the onset of ethylene-related ripening in Malus × domestica (apples) at both the hormone and transcriptome levels. Methodology Transgenic apples suppressed for the SEPALLATA1/2 (SEP1/2) class of gene (MADS8/9) that showed severely reduced ripening were compared with untransformed control apples. In each apple type, free indole-3-acetic acid (IAA) concentrations were measured during early ripening. The changes observed in auxin were assessed in light of global changes in gene expression. Principal results It was found that mature MADS8/9-suppressed apples had a higher concentration of free IAA. This was associated with increased expression of the auxin biosynthetic genes in the indole-3-acetamide pathway. Additionally, in the MADS8/9-suppressed apples, there was less expression of the GH3 auxin-conjugating enzymes. A number of genes involved in the auxin-regulated transcription (AUX/IAA and ARF classes of genes) were also observed to change in expression, suggesting a mechanism for signal transduction at the start of ripening. Conclusions The delay in ripening observed in MADS8/9-suppressed apples may be partly due to high auxin concentrations. We propose that, to achieve low auxin associated with fruit maturation, the auxin homeostasis is controlled in a two-pronged manner: (i) by the reduction in biosynthesis and (ii) by an increase in auxin conjugation. This is associated with the change in expression of auxin-signalling genes and the up-regulation of ripening-related genes. PMID:23346344

  18. Investigation into the role of endogenous abscisic acid during ripening of imported avocado cv. Hass.

    PubMed

    Meyer, Marjolaine D; Chope, Gemma A; Terry, Leon A

    2017-08-01

    The importance of ethylene in avocado ripening has been extensively studied. In contrast, little is known about the possible role of abscisic acid (ABA). The present work studied the effect of 1-methylcyclopropene (1-MCP) (0.3 μL L -1 ), e+® Ethylene Remover and the combination thereof on the quality of imported avocado cv. Hass fruit stored for 7 days at 12 °C. Ethylene production, respiration, firmness, colour, heptose (C7) sugars and ABA concentrations in mesocarp tissue were measured throughout storage. Treatment with e+® Ethylene Remover reduced ethylene production, respiration rate and physiological ripening compared with controls. Fruit treated with 1-MCP + e+® Ethylene Remover and, to a lesser extent 1-MCP alone, had the lowest ethylene production and respiration rate and hence the best quality. Major sugars measured in mesocarp tissue were mannoheptulose and perseitol, and their content was not correlated with ripening parameters. Mesocarp ABA concentration, as determined by mass spectrometry, increased as fruit ripened and was negatively correlated with fruit firmness. Results suggest a relationship between ABA and ethylene metabolism since blocking ethylene, and to a larger extent blocking and removing ethylene, resulted in lower ABA concentrations. Whether ABA influences avocado fruit ripening needs to be determined in future research. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  19. Fibroblast Growth Factor 2: An Epithelial Ductal Cell Growth Inhibitor That Drops Out in Breast Cancer

    DTIC Science & Technology

    2009-10-01

    AD_________________ Award Number: W81XWH-08-1-0708 TITLE: Fibroblast Growth Factor 2: an...September 2008 – 14 September 2009 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Fibroblast Growth Factor 2: an Epithelial Ductal Cell Growth Inhibitor...9 Fibroblast Growth Factor -2: an Epithelial Ductal Cell Growth Inhibitor that Drops Out in Breast Cancer

  20. Spatial Distribution of Lactococcus lactis Colonies Modulates the Production of Major Metabolites during the Ripening of a Model Cheese.

    PubMed

    Le Boucher, Clémentine; Gagnaire, Valérie; Briard-Bion, Valérie; Jardin, Julien; Maillard, Marie-Bernadette; Dervilly-Pinel, Gaud; Le Bizec, Bruno; Lortal, Sylvie; Jeanson, Sophie; Thierry, Anne

    2016-01-01

    In cheese, lactic acid bacteria are immobilized at the coagulation step and grow as colonies. The spatial distribution of bacterial colonies is characterized by the size and number of colonies for a given bacterial population within cheese. Our objective was to demonstrate that different spatial distributions, which lead to differences in the exchange surface between the colonies and the cheese matrix, can influence the ripening process. The strategy was to generate cheeses with the same growth and acidification of a Lactococcus lactis strain with two different spatial distributions, big and small colonies, to monitor the production of the major ripening metabolites, including sugars, organic acids, peptides, free amino acids, and volatile metabolites, over 1 month of ripening. The monitored metabolites were qualitatively the same for both cheeses, but many of them were more abundant in the small-colony cheeses than in the big-colony cheeses over 1 month of ripening. Therefore, the results obtained showed that two different spatial distributions of L. lactis modulated the ripening time course by generating moderate but significant differences in the rates of production or consumption for many of the metabolites commonly monitored throughout ripening. The present work further explores the immobilization of bacteria as colonies within cheese and highlights the consequences of this immobilization on cheese ripening. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  1. Role of the Tomato Non-Ripening Mutation in Regulating Fruit Quality Elucidated Using iTRAQ Protein Profile Analysis

    PubMed Central

    Yuan, Xin-Yu; Wang, Rui-Heng; Zhao, Xiao-Dan; Luo, Yun-Bo; Fu, Da-Qi

    2016-01-01

    Natural mutants of the Non-ripening (Nor) gene repress the normal ripening of tomato fruit. The molecular mechanism of fruit ripening regulation by the Nor gene is unclear. To elucidate how the Nor gene can affect ripening and fruit quality at the protein level, we used the fruits of Nor mutants and wild-type Ailsa Craig (AC) to perform iTRAQ (isobaric tags for relative and absolute quantitation) analysis. The Nor mutation altered tomato fruit ripening and affected quality in various respects, including ethylene biosynthesis by down-regulating the abundance of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO), pigment biosynthesis by repressing phytoene synthase 1 (PSY1), ζ-carotene isomerase (Z-ISO), chalcone synthase 1 (CHS1) and other proteins, enhancing fruit firmness by increasing the abundance of cellulose synthase protein, while reducing those of polygalacturonase 2 (PG2) and pectate lyase (PL), altering biosynthesis of nutrients such as carbohydrates, amino acids, and anthocyanins. Conversely, Nor mutation also enhanced the fruit’s resistance to some pathogens by up-regulating the expression of several genes associated with stress and defense. Therefore, the Nor gene is involved in the regulation of fruit ripening and quality. It is useful in the future as a means to improve fruit quality in tomato. PMID:27732677

  2. Physiological and proteomic approaches to address the active role of ozone in kiwifruit post-harvest ripening

    PubMed Central

    Minas, Ioannis S.; Tanou, Georgia; Belghazi, Maya; Job, Dominique; Manganaris, George A.; Molassiotis, Athanassios; Vasilakakis, Miltiadis

    2012-01-01

    Post-harvest ozone application has recently been shown to inhibit the onset of senescence symptoms on fleshy fruit and vegetables; however, the exact mechanism of action is yet unknown. To characterize the impact of ozone on the post-harvest performance of kiwifruit (Actinidia deliciosa cv. ‘Hayward’), fruits were cold stored (0 °C, 95% relative humidity) in a commercial ethylene-free room for 1, 3, or 5 months in the absence (control) or presence of ozone (0.3 μl l−1) and subsequently were allowed to ripen at a higher temperature (20 °C), herein defined as the shelf-life period, for up to 12 days. Ozone blocked ethylene production, delayed ripening, and stimulated antioxidant and anti-radical activities of fruits. Proteomic analysis using 1D-SDS-PAGE and mass spectrometry identified 102 kiwifruit proteins during ripening, which are mainly involved in energy, protein metabolism, defence, and cell structure. Ripening induced protein carbonylation in kiwifruit but this effect was depressed by ozone. A set of candidate kiwifruit proteins that are sensitive to carbonylation was also discovered. Overall, the present data indicate that ozone improved kiwifruit post-harvest behaviour, thus providing a first step towards understanding the active role of this molecule in fruit ripening. PMID:22268155

  3. An infrared based sensor system for the detection of ethylene for the discrimination of fruit ripening

    NASA Astrophysics Data System (ADS)

    Kathirvelan, J.; Vijayaraghavan, R.

    2017-09-01

    We report the fabrication and testing of a prototype ethylene sensing device for use in fruit ripening applications. A sensor based on infrared (IR) thermal emission was developed and used to detect the ethylene level released during the fruit ripening process. An IR thermal source tuned to the 10.6 μm wavelength was linked to a high-sensitivity silicon temperature detector. When introduced into the wave path between the IR source and temperature detector, ethylene absorbs the 10.6 μm IR waves and decreases the surface temperature of the detector. The output is then converted to an electrical signal (in mV), which gives a direct measurement of the ethylene level. Using this sensor, ethylene concentration measured from a fruit sample continuously decreased from 59 to 5 ppm during the natural ripening process. The sensor exhibited a sensitivity of 3.3 ± 0.2% (change in detector output (mV)/ppm × 100) and could measure concentrations as low as 5 ppm with rise and recovery times of 1 and 3 s, respectively. The system demonstrated good reproducibility. Devices employing this sensor system may be used for fruit ripening applications on site and in the field and for screening artificially ripened fruits, therefore contributing to ensure food safety.

  4. Transcriptome analysis during ripening of table grape berry cv. Thompson Seedless.

    PubMed

    Balic, Iván; Vizoso, Paula; Nilo-Poyanco, Ricardo; Sanhueza, Dayan; Olmedo, Patricio; Sepúlveda, Pablo; Arriagada, Cesar; Defilippi, Bruno G; Meneses, Claudio; Campos-Vargas, Reinaldo

    2018-01-01

    Ripening is one of the key processes associated with the development of major organoleptic characteristics of the fruit. This process has been extensively characterized in climacteric fruit, in contrast with non-climacteric fruit such as grape, where the process is less understood. With the aim of studying changes in gene expression during ripening of non-climacteric fruit, an Illumina based RNA-Seq transcriptome analysis was performed on four developmental stages, between veraison and harvest, on table grapes berries cv Thompson Seedless. Functional analysis showed a transcriptional increase in genes related with degradation processes of chlorophyll, lipids, macromolecules recycling and nucleosomes organization; accompanied by a decrease in genes related with chloroplasts integrity and amino acid synthesis pathways. It was possible to identify several processes described during leaf senescence, particularly close to harvest. Before this point, the results suggest a high transcriptional activity associated with the regulation of gene expression, cytoskeletal organization and cell wall metabolism, which can be related to growth of berries and firmness loss characteristic to this stage of development. This high metabolic activity could be associated with an increase in the transcription of genes related with glycolysis and respiration, unexpected for a non-climacteric fruit ripening.

  5. Evolutionary Recycling of Light Signaling Components in Fleshy Fruits: New Insights on the Role of Pigments to Monitor Ripening

    PubMed Central

    Llorente, Briardo; D’Andrea, Lucio; Rodríguez-Concepción, Manuel

    2016-01-01

    Besides an essential source of energy, light provides environmental information to plants. Photosensory pathways are thought to have occurred early in plant evolution, probably at the time of the Archaeplastida ancestor, or perhaps even earlier. Manipulation of individual components of light perception and signaling networks in tomato (Solanum lycopersicum) affects the metabolism of ripening fruit at several levels. Most strikingly, recent experiments have shown that some of the molecular mechanisms originally devoted to sense and respond to environmental light cues have been re-adapted during evolution to provide plants with useful information on fruit ripening progression. In particular, the presence of chlorophylls in green fruit can strongly influence the spectral composition of the light filtered through the fruit pericarp. The concomitant changes in light quality can be perceived and transduced by phytochromes (PHYs) and PHY-interacting factors, respectively, to regulate gene expression and in turn modulate the production of carotenoids, a family of metabolites that are relevant for the final pigmentation of ripe fruits. We raise the hypothesis that the evolutionary recycling of light-signaling components to finely adjust pigmentation to the actual ripening stage of the fruit may have represented a selective advantage for primeval fleshy-fruited plants even before the extinction of dinosaurs. PMID:27014289

  6. Evolutionary Recycling of Light Signaling Components in Fleshy Fruits: New Insights on the Role of Pigments to Monitor Ripening.

    PubMed

    Llorente, Briardo; D'Andrea, Lucio; Rodríguez-Concepción, Manuel

    2016-01-01

    Besides an essential source of energy, light provides environmental information to plants. Photosensory pathways are thought to have occurred early in plant evolution, probably at the time of the Archaeplastida ancestor, or perhaps even earlier. Manipulation of individual components of light perception and signaling networks in tomato (Solanum lycopersicum) affects the metabolism of ripening fruit at several levels. Most strikingly, recent experiments have shown that some of the molecular mechanisms originally devoted to sense and respond to environmental light cues have been re-adapted during evolution to provide plants with useful information on fruit ripening progression. In particular, the presence of chlorophylls in green fruit can strongly influence the spectral composition of the light filtered through the fruit pericarp. The concomitant changes in light quality can be perceived and transduced by phytochromes (PHYs) and PHY-interacting factors, respectively, to regulate gene expression and in turn modulate the production of carotenoids, a family of metabolites that are relevant for the final pigmentation of ripe fruits. We raise the hypothesis that the evolutionary recycling of light-signaling components to finely adjust pigmentation to the actual ripening stage of the fruit may have represented a selective advantage for primeval fleshy-fruited plants even before the extinction of dinosaurs.

  7. Photopyroelectric Monitoring of Olive's Ripening Conditions and Olive Oil Quality Using Pulsed Wideband IR Thermal Source

    NASA Astrophysics Data System (ADS)

    Abu-Taha, M. I.; Sarahneh, Y.; Saleh, A. M.

    The present study is based on band absorption of radiation from pulsed wideband infrared (IR) thermal source (PWBS) in conjunction with polyvinylidene fluoride film (PVDF). It is the first time to be employed to monitor the ripening state of olive fruit. Olive's characteristics vary at different stages of ripening, and hence, cultivation of olives at the right time is important in ensuring the best oil quality and maximizes the harvest yield. The photopyroelectric (PPE) signal resulting from absorption of wideband infrared (IR) radiation by fresh olive juice indicates the ripening stage of olives, i.e., allows an estimate of the suitable harvest time. The technique was found to be very useful in discriminating between olive oil samples according to geographical region, shelf life, some storage conditions, and deliberate adulteration. Our results for monitoring oil accumulation in olives during the ripening season agree well with the complicated analytical studies carried out by other researchers.

  8. Ripening of pepper (Capsicum annuum) fruit is characterized by an enhancement of protein tyrosine nitration

    PubMed Central

    Chaki, Mounira; Álvarez de Morales, Paz; Ruiz, Carmelo; Begara-Morales, Juan C.; Barroso, Juan B.; Corpas, Francisco J.; Palma, José M.

    2015-01-01

    Background and Aims Pepper (Capsicum annuum, Solanaceae) fruits are consumed worldwide and are of great economic importance. In most species ripening is characterized by important visual and metabolic changes, the latter including emission of volatile organic compounds associated with respiration, destruction of chlorophylls, synthesis of new pigments (red/yellow carotenoids plus xanthophylls and anthocyanins), formation of pectins and protein synthesis. The involvement of nitric oxide (NO) in fruit ripening has been established, but more work is needed to detail the metabolic networks involving NO and other reactive nitrogen species (RNS) in the process. It has been reported that RNS can mediate post-translational modifications of proteins, which can modulate physiological processes through mechanisms of cellular signalling. This study therefore examined the potential role of NO in nitration of tyrosine during the ripening of California sweet pepper. Methods The NO content of green and red pepper fruit was determined spectrofluorometrically. Fruits at the breaking point between green and red coloration were incubated in the presence of NO for 1 h and then left to ripen for 3 d. Profiles of nitrated proteins were determined using an antibody against nitro-tyrosine (NO2-Tyr), and profiles of nitrosothiols were determined by confocal laser scanning microscopy. Nitrated proteins were identified by 2-D electrophoresis and MALDI-TOF/TOF analysis. Key Results Treatment with NO delayed the ripening of fruit. An enhancement of nitrosothiols and nitroproteins was observed in fruit during ripening, and this was reversed by the addition of exogenous NO gas. Six nitrated proteins were identified and were characterized as being involved in redox, protein, carbohydrate and oxidative metabolism, and in glutamate biosynthesis. Catalase was the most abundant nitrated protein found in both green and red fruit. Conclusions The RNS profile reported here indicates that ripening of

  9. Functional genomics provides insights into the role of Propionibacterium freudenreichii ssp. shermanii JS in cheese ripening.

    PubMed

    Ojala, Teija; Laine, Pia K S; Ahlroos, Terhi; Tanskanen, Jarna; Pitkänen, Saara; Salusjärvi, Tuomas; Kankainen, Matti; Tynkkynen, Soile; Paulin, Lars; Auvinen, Petri

    2017-01-16

    Propionibacterium freudenreichii is a commercially important bacterium that is essential for the development of the characteristic eyes and flavor of Swiss-type cheeses. These bacteria grow actively and produce large quantities of flavor compounds during cheese ripening at warm temperatures but also appear to contribute to the aroma development during the subsequent cold storage of cheese. Here, we advance our understanding of the role of P. freudenreichii in cheese ripening by presenting the 2.68-Mbp annotated genome sequence of P. freudenreichii ssp. shermanii JS and determining its global transcriptional profiles during industrial cheese-making using transcriptome sequencing. The annotation of the genome identified a total of 2377 protein-coding genes and revealed the presence of enzymes and pathways for formation of several flavor compounds. Based on transcriptome profiling, the expression of 348 protein-coding genes was altered between the warm and cold room ripening of cheese. Several propionate, acetate, and diacetyl/acetoin production related genes had higher expression levels in the warm room, whereas a general slowing down of the metabolism and an activation of mobile genetic elements was seen in the cold room. A few ripening-related and amino acid catabolism involved genes were induced or remained active in cold room, indicating that strain JS contributes to the aroma development also during cold room ripening. In addition, we performed a comparative genomic analysis of strain JS and 29 other Propionibacterium strains of 10 different species, including an isolate of both P. freudenreichii subspecies freudenreichii and shermanii. Ortholog grouping of the predicted protein sequences revealed that close to 86% of the ortholog groups of strain JS, including a variety of ripening-related ortholog groups, were conserved across the P. freudenreichii isolates. Taken together, this study contributes to the understanding of the genomic basis of P. freudenreichii

  10. Performance of two alternative methods for Listeria detection throughout Serro Minas cheese ripening.

    PubMed

    Mata, Gardênia Márcia Silva Campos; Martins, Evandro; Machado, Solimar Gonçalves; Pinto, Maximiliano Soares; de Carvalho, Antônio Fernandes; Vanetti, Maria Cristina Dantas

    2016-01-01

    The ability of pathogens to survive cheese ripening is a food-security concern. Therefore, this study aimed to evaluate the performance of two alternative methods of analysis of Listeria during the ripening of artisanal Minas cheese. These methods were tested and compared with the conventional method: Lateral Flow System™, in cheeses produced on laboratory scale using raw milk collected from different farms and inoculated with Listeria innocua; and VIDAS(®)-LMO, in cheese samples collected from different manufacturers in Serro, Minas Gerais, Brazil. These samples were also characterized in terms of lactic acid bacteria, coliforms and physical-chemical analysis. In the inoculated samples, L. innocua was detected by Lateral Flow System™ method with 33% false-negative and 68% accuracy results. L. innocua was only detected in the inoculated samples by the conventional method at 60-days of cheese ripening. L. monocytogenes was not detected by the conventional and the VIDAS(®)-LMO methods in cheese samples collected from different manufacturers, which impairs evaluating the performance of this alternative method. We concluded that the conventional method provided a better recovery of L. innocua throughout cheese ripening, being able to detect L. innocua at 60-day, aging period which is required by the current legislation. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  11. Regulation of Wheat Seed Dormancy by After-Ripening Is Mediated by Specific Transcriptional Switches That Induce Changes in Seed Hormone Metabolism and Signaling

    PubMed Central

    Kanno, Yuri; Jordan, Mark C.; Kamiya, Yuji; Seo, Mitsunori; Ayele, Belay T.

    2013-01-01

    Treatments that promote dormancy release are often correlated with changes in seed hormone content and/or sensitivity. To understand the molecular mechanisms underlying the role of after-ripening (seed dry storage) in triggering hormone related changes and dormancy decay in wheat (Triticum aestivum), temporal expression patterns of genes related to abscisic acid (ABA), gibberellin (GA), jasmonate and indole acetic acid (IAA) metabolism and signaling, and levels of the respective hormones were examined in dormant and after-ripened seeds in both dry and imbibed states. After-ripening mediated developmental switch from dormancy to germination appears to be associated with declines in seed sensitivity to ABA and IAA, which are mediated by transcriptional repressions of PROTEIN PHOSPHATASE 2C, SNF1-RELATED PROTEIN KINASE2, ABA INSENSITIVE5 and LIPID PHOSPHATE PHOSPHTASE2, and AUXIN RESPONSE FACTOR and RELATED TO UBIQUITIN1 genes. Transcriptomic analysis of wheat seed responsiveness to ABA suggests that ABA inhibits the germination of wheat seeds partly by repressing the transcription of genes related to chromatin assembly and cell wall modification, and activating that of GA catabolic genes. After-ripening induced seed dormancy decay in wheat is also associated with the modulation of seed IAA and jasmonate contents. Transcriptional control of members of the ALLENE OXIDE SYNTHASE, 3-KETOACYL COENZYME A THIOLASE, LIPOXYGENASE and 12-OXOPHYTODIENOATE REDUCTASE gene families appears to regulate seed jasmonate levels. Changes in the expression of GA biosynthesis genes, GA 20-OXIDASE and GA 3-OXIDASE, in response to after-ripening implicate this hormone in enhancing dormancy release and germination. These findings have important implications in the dissection of molecular mechanisms underlying regulation of seed dormancy in cereals. PMID:23437172

  12. Role of Sucrose Phosphate Synthase in Sucrose Biosynthesis in Ripening Bananas and Its Relationship to the Respiratory Climacteric 1

    PubMed Central

    Hubbard, Natalie L.; Pharr, D. Mason; Huber, Steven C.

    1990-01-01

    During ripening of bananas (Musa spp. [AAA group, Cavendish subgroup]), there is a massive conversion of starch to sucrose. Also during ripening there is a rise in respiration known as the respiratory climacteric. In this study changes in carbohydrate content, activities of starch and sucrose metabolizing enzymes, and respiration were measured to assess their potential interrelationships. Sucrose phosphate synthase activity increased dramatically during the first 4 days after initiation of ripening by ethylene treatment. Starch concentration decreased and sucrose concentration increased during this time period. Developmental changes in sucrose phosphate synthase activity were measured with limiting substrate (plus Pi) and saturating substrate concentrations. Activities were not parallel under the two assay conditions, providing tentative evidence that kinetically different forms of the enzyme may exist at different stages of ripening. Sucrose accumulation rate was most highly correlated with sucrose phosphate synthase activity assayed with limiting substrate concentrations (plus Pi). The cumulative amount of CO2 respired during ripening was positively correlated with sugar accumulation (R2 = 0.97). From this linear regression it was calculated that a constant 0.605 millimoles of CO2 was evolved per mole of sucrose formed throughout ripening. Using this quantity, the percentage of the total respiratory ATP produced which was required for the conversion of starch to sucrose was calculated assuming different models for carbon export from the amyloplast. The results suggest that sucrose biosynthesis during ripening constitutes a significant sink for respiratory ATP. PMID:16667688

  13. PacCYP707A2 negatively regulates cherry fruit ripening while PacCYP707A1 mediates drought tolerance

    PubMed Central

    Li, Qian; Chen, Pei; Dai, Shengjie; Sun, Yufei; Yuan, Bing; Kai, Wenbin; Pei, Yuelin; He, Suihuan; Liang, Bin; Zhang, Yushu; Leng, Ping

    2015-01-01

    Sweet cherry is a non-climacteric fruit and its ripening is regulated by abscisic acid (ABA) during fruit development. In this study, four cDNAs (PacCYP707A1–4) encoding 8′-hydroxylase, a key enzyme in the oxidative catabolism of ABA, were identified in sweet cherry fruits using tobacco rattle virus-induced gene silencing (VIGS) and particle bombardment approaches. Quantitative real-time PCR confirmed significant down-regulation of target gene transcripts in VIGS-treated cherry fruits. In PacCYP707A2-RNAi-treated fruits, ripening and fruit colouring were promoted relative to control fruits, and both ABA accumulation and PacNCED1 transcript levels were up-regulated by 140%. Silencing of PacCYP707A2 by VIGS significantly altered the transcripts of both ABA-responsive and ripening-related genes, including the ABA metabolism-associated genes NCED and CYP707A, the anthocyanin synthesis genes PacCHS, PacCHI, PacF3H, PacDFR, PacANS, and PacUFGT, the ethylene biosynthesis gene PacACO1, and the transcription factor PacMYBA. The promoter of PacMYBA responded more strongly to PacCYP707A2-RNAi-treated fruits than to PacCYP707A1-RNAi-treated fruits. By contrast, silencing of PacCYP707A1 stimulated a slight increase in fruit colouring and enhanced resistance to dehydration stress compared with control fruits. These results suggest that PacCYP707A2 is a key regulator of ABA catabolism that functions as a negative regulator of fruit ripening, while PacCYP707A1 regulates ABA content in response to dehydration during fruit development. PMID:25956880

  14. Tissue Factor Pathway Inhibitor: Multiple Anticoagulant Activities for a Single Protein.

    PubMed

    Mast, Alan E

    2016-01-01

    Tissue factor (TF) pathway inhibitor (TFPI) is an anticoagulant protein that inhibits early phases of the procoagulant response. Alternatively spliced isoforms of TFPI are differentially expressed by endothelial cells and human platelets and plasma. The TFPIβ isoform localizes to the endothelium surface where it is a potent inhibitor of TF-factor VIIa complexes that initiate blood coagulation. The TFPIα isoform is present in platelets. TFPIα contains a stretch of 9 amino acids nearly identical to those found in the B-domain of factor V that are well conserved in mammals. These amino acids provide exosite binding to activated factor V, which allows for TFPIα to inhibit prothrombinase during the initiation phase of blood coagulation. Endogenous inhibition at this point in the coagulation cascade was only recently recognized and has provided a biochemical rationale to explain the pathophysiological mechanisms underlying several clinical disorders. These include the east Texas bleeding disorder that is caused by production of an altered form of factor V with high affinity for TFPI and a paradoxical procoagulant effect of heparins. In addition, these findings have led to ideas for pharmacological targeting of TFPI that may reduce bleeding in hemophilia patients. © 2015 American Heart Association, Inc.

  15. Microbial diversity and dynamics throughout manufacturing and ripening of surface ripened semi-hard Danish Danbo cheeses investigated by culture-independent techniques.

    PubMed

    Ryssel, Mia; Johansen, Pernille; Al-Soud, Waleed Abu; Sørensen, Søren; Arneborg, Nils; Jespersen, Lene

    2015-12-23

    Microbial successions on the surface and in the interior of surface ripened semi-hard Danish Danbo cheeses were investigated by culture-dependent and -independent techniques. Culture-independent detection of microorganisms was obtained by denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, using amplicons of 16S and 26S rRNA genes for prokaryotes and eukaryotes, respectively. With minor exceptions, the results from the culture-independent analyses correlated to the culture-dependent plating results. Even though the predominant microorganisms detected with the two culture-independent techniques correlated, a higher number of genera were detected by pyrosequencing compared to DGGE. Additionally, minor parts of the microbiota, i.e. comprising <10.0% of the operational taxonomic units (OTUs), were detected by pyrosequencing, resulting in more detailed information on the microbial succession. As expected, microbial profiles of the surface and the interior of the cheeses diverged. During cheese production pyrosequencing determined Lactococcus as the dominating genus on cheese surfaces, representing on average 94.7%±2.1% of the OTUs. At day 6 Lactococcus spp. declined to 10.0% of the OTUs, whereas Staphylococcus spp. went from 0.0% during cheese production to 75.5% of the OTUs at smearing. During ripening, i.e. from 4 to 18 weeks, Corynebacterium was the dominant genus on the cheese surface (55.1%±9.8% of the OTUs), with Staphylococcus (17.9%±11.2% of the OTUs) and Brevibacterium (10.4%±8.3% of the OTUs) being the second and third most abundant genera. Other detected bacterial genera included Clostridiisalibacter (5.0%±4.0% of the OTUs), as well as Pseudoclavibacter, Alkalibacterium and Marinilactibacillus, which represented <2% of the OTUs. At smearing, yeast counts were low with Debaryomyces being the dominant genus accounting for 46.5% of the OTUs. During ripening the yeast counts increased significantly with Debaryomyces being the predominant genus

  16. Synthesis and P1' SAR exploration of potent macrocyclic tissue factor-factor VIIa inhibitors

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ladziata, Vladimir; Glunz, Peter W.; Zou, Yan

    Selective tissue factor-factor VIIa complex (TF-FVIIa) inhibitors are viewed as promising compounds for treating thrombotic disease. In this contribution, we describe multifaceted exploratory SAR studies of S1'-binding moieties within a macrocyclic chemotype aimed at replacing cyclopropyl sulfone P1' group. Over the course of the optimization efforts, the 1-(1H-tetrazol-5-yl)cyclopropane P1' substituent emerged as an improved alternative, offering increased metabolic stability and lower clearance, while maintaining excellent potency and selectivity.

  17. Proteomic analysis of ripening tomato fruit infected by Botrytis cinerea.

    PubMed

    Shah, Punit; Powell, Ann L T; Orlando, Ron; Bergmann, Carl; Gutierrez-Sanchez, Gerardo

    2012-04-06

    Botrytis cinerea, a model necrotrophic fungal pathogen that causes gray mold as it infects different organs on more than 200 plant species, is a significant contributor to postharvest rot in fresh fruit and vegetables, including tomatoes. By describing host and pathogen proteomes simultaneously in infected tissues, the plant proteins that provide resistance and allow susceptibility and the pathogen proteins that promote colonization and facilitate quiescence can be identified. This study characterizes fruit and fungal proteins solubilized in the B. cinerea-tomato interaction using shotgun proteomics. Mature green, red ripe wild type and ripening inhibited (rin) mutant tomato fruit were infected with B. cinerea B05.10, and the fruit and fungal proteomes were identified concurrently 3 days postinfection. One hundred eighty-six tomato proteins were identified in common among red ripe and red ripe-equivalent ripening inhibited (rin) mutant tomato fruit infected by B. cinerea. However, the limited infections by B. cinerea of mature green wild type fruit resulted in 25 and 33% fewer defense-related tomato proteins than in red and rin fruit, respectively. In contrast, the ripening stage of genotype of the fruit infected did not affect the secreted proteomes of B. cinerea. The composition of the collected proteins populations and the putative functions of the identified proteins argue for their role in plant-pathogen interactions.

  18. Sexual intercourse for cervical ripening and induction of labour.

    PubMed

    Kavanagh, J; Kelly, A J; Thomas, J

    2001-01-01

    The role of prostaglandins for cervical ripening and induction of labour has been examined extensively. Human semen is the biological source that is presumed to contain the highest prostaglandin concentration. The role of sexual intercourse in the initiation of labour is uncertain. The action of sexual intercourse in stimulating labour is unclear, it may in part be due to the physical stimulation of the lower uterine segment, or endogenous release of oxytocin as a result of orgasm or from the direct action of prostaglandins in semen. Furthermore nipple stimulation may be part of the process of initiation. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. To determine the effects of sexual intercourse for third trimester cervical ripening or induction of labour in comparison with other methods of induction. The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies of relevant papers. Last searched: November 2000. (1) clinical trials comparing sexual intercourse for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusion. A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involves a two-stage method of data extraction. There was one included study of 28 women which reported very limited data, from which no meaningful conclusions can be drawn. The role of sexual intercourse as a method of induction

  19. Biochemistry and Cell Wall Changes Associated with Noni (Morinda citrifolia L.) Fruit Ripening.

    PubMed

    Cárdenas-Coronel, Wendy G; Carrillo-López, Armando; Vélez de la Rocha, Rosabel; Labavitch, John M; Báez-Sañudo, Manuel A; Heredia, José B; Zazueta-Morales, José J; Vega-García, Misael O; Sañudo-Barajas, J Adriana

    2016-01-13

    Quality and compositional changes were determined in noni fruit harvested at five ripening stages, from dark-green to thaslucent-grayish. Fruit ripening was accompanied by acidity and soluble solids accumulation but pH diminution, whereas the softening profile presented three differential steps named early (no significant softening), intermediate (significant softening), and final (dramatic softening). At early step the extensive depolymerization of hydrosoluble pectins and the significantly increment of pectinase activities did not correlate with the slight reduction in firmness. The intermediate step showed an increment of pectinases and hemicellulases activities. The final step was accompanied by the most significant reduction in the yield of alcohol-insoluble solids as well as in the composition of uronic acids and neutral sugars; pectinases increased their activity and depolymerization of hemicellulosic fractions occurred. Noni ripening is a process conducted by the coordinated action of pectinases and hemicellulases that promote the differential dissasembly of cell wall polymers.

  20. Mannosyl- and Xylosyl-Containing Glycans Promote Tomato (Lycopersicon esculentum Mill.) Fruit Ripening

    PubMed Central

    Priem, Bernard; Gross, Kenneth C.

    1992-01-01

    The oligosaccharide glycans mannosylα1-6(mannosylα1-3)mannosylα1-6(mannosylα1-3) mannosylβ1-4-N-acetylglucosamine and mannosylα1-6(mannosylα1-3)(xylosylβ1-2) mannosylβ1-4-N-acetylglucosaminyl(fucosylα1-3) N-acetylglucosamine were infiltrated into mature green tomato fruit (Lycopersicon esculentum Mill., cv Rutgers). Coinfiltration of 1 nanogram per gram fresh weight of the glycans with 40 micrograms per gram fresh weight galactose, a level of galactose insufficient to promote ripening, stimulated ripening as measured by red coloration and ethylene production. PMID:16668644

  1. Expression of a ripening-related cytochrome P450 cDNA in Cavendish banana (Musa acuminata cv. Williams).

    PubMed

    Pua, Eng-Chong; Lee, Yi-Chuan

    2003-02-13

    As part of a study to understand the molecular basis of fruit ripening, this study reports the isolation and characterization of a banana cytochrome P450 (P450) cDNA, designated as MAP450-1, which was associated with fruit ripening of banana. MAP450-1 encoded a single polypeptide of 507 amino acid residues that shared an overall identity of 27-45% with that of several plant P450s, among which MAP450-1 was most related phylogenetically to the avocado P450 CYP71A1. The polypeptide that possessed residue domains conserved in all P450s was classified as CYP71N1. Expression of CYP71N1 varied greatly between banana organs. Transcripts were detected only in peel and pulp of the ripening fruit and not in unripe fruit tissues at all developmental stages or other organs (root, leaf, ovary and flower). During ripening, transcripts were barely detectable in pre-climacteric and climacteric fruits but, as ripening progressed, they began to accumulate and reached a maximum in post-climacteric fruits. CYP71N1 expression in pre-climacteric fruit could be upregulated by exogenous application of ethylene (1-5 ppm) and treatment of overripe fruit with exogenous sucrose (50-300 mM) but not glucose downregulated the expression. These results indicate that P450s may not play a role in fruit development and its expression is associated with ripening, which may be regulated, in part, by ethylene and/or sucrose, at the transcript level.

  2. Nitric oxide donors for cervical ripening and induction of labour.

    PubMed

    Kelly, Anthony J; Munson, Christopher; Minden, Lucy

    2011-06-15

    Sometimes it is necessary to bring on labour artificially because of safety concerns for the mother or baby. This review is one of a series of reviews of methods of labour induction using a standardised protocol.Induction of labour occurs in approximately 20% of pregnancies in the UK. The ideal agent for induction of labour would induce cervical ripening without causing uterine contractions. Currently most commonly used cervical ripening or induction agents result in uterine activity or contractions, or both. Cervical ripening without uterine contractility could occur safely in an outpatient setting and it may be expected that this would result in greater maternal satisfaction and lower costs. To determine the effects of nitric oxide (NO) donors for third trimester cervical ripening or induction of labour, in comparison with placebo or no treatment or other treatments from a predefined hierarchy. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (31 December 2010) and the reference lists of trial reports and reviews. Clinical trials comparing NO donors for cervical ripening or labour induction to other methods listed above it on a predefined list of methods of labour induction. The trials include some form of random allocation to either group; and report one or more of the prestated outcomes. NO donors (isosorbide mononitrate, nitroglycerin and sodium nitroprusside) are compared to other methods listed above it on a predefined list of methods of labour induction. This review is part of a series of reviews focusing on methods of induction of labour. Three review authors independently assessed trials for inclusion, assessed risk of bias and extracted data. We considered 19 trials; we included 10 (including a total of 1889 women) trials, excluded eight trials and one trial report is awaiting classification. Included studies compared NO donors to placebo, vaginal prostaglandin E2, intracervical PGE2 and vaginal misoprostol. All included studies

  3. Pinning Stabilizes Neighboring Surface Nanobubbles against Ostwald Ripening.

    PubMed

    Dollet, Benjamin; Lohse, Detlef

    2016-11-01

    Pinning of the contact line and gas oversaturation explain the stability of single surface nanobubbles. In this article, we theoretically show that the pinning also suppresses the Ostwald ripening process between neighboring surface nanobubbles, thus explaining why in a population of neighboring surface nanobubbles different radii of curvature of the nanobubbles can be observed.

  4. Risk Factors for Inhibitor Formation in Hemophilia: A Prevalent Case-Control Study

    PubMed Central

    Ragni, Margaret V.; Ojeifo, Oluseyi; Feng, Jinong; Yan, Jin; Hill, Kathleen A.; Sommer, Steve S.; Trucco, Massimo N.; Brambilla, Donald J.

    2009-01-01

    Background Inhibitor formation is a major complication of hemophilia treatment. Aim In a prevalent case-control study, we evaluated blood product exposure, genotype, and HLA type on hemophilia A inhibitor formation. Methods Product exposure was extracted from medical records. Genotype was determined on stored DNA samples by detection of virtually all mutations-SSCP (DOVAM-S) and subcycling PCR. HLA typing was performed by PCR amplification and exonuclease-released fluorescence. Results Cases experienced higher intensity factor, 455 vs. 200 U per exposure, p<0.005, more frequent central nervous system (CNS) bleeding, 7 of 20 (35.0%) vs. 1 of 57 (1.7%), p=0.001, and more commonly from inhibitor families, 7 of 20 (35.0%) vs. 0 of 57 (0%), p<0.001, and African-American, 12 of 63 (19.0%) vs. 6 of 117 (5.1%), p=0.015. Among the latter, CNS bleeding was more commonly the initial bleed, 60% vs. 0%, p<0.001, and survival was shorter, 14 vs. 38 yr, p=0.025. Inhibitor formation was uncommon in those with missense mutations, 2 of 65 (3.1%) vs. 31 of 119 (26.0%), p=0.008, and unrelated to factor VIII immunogenic epitope, p=0.388, or HLA type, p>0.100. Genotype was not associated with race. Time to immune tolerance was shorter for titers < 120 vs. ≥ 120 BU/ml, 6 vs. 16 months, p<0.01, but unaffected by tolerizing dose regimen, p>0.50. Conclusions Inhibitor formation is associated with high intensity product exposure, CNS bleeding, African-American race, and low frequency of missense mutations. The ideal time to initiate prophylaxis to reduce CNS bleeding and inhibitor formation will require prospective studies. PMID:19563499

  5. Monitoring of compositional changes during berry ripening in grape seed extracts of cv. Sangiovese (Vitis vinifera L.).

    PubMed

    Bombai, Giuseppe; Pasini, Federica; Verardo, Vito; Sevindik, Onur; Di Foggia, Michele; Tessarin, Paola; Bregoli, Anna Maria; Caboni, Maria F; Rombolà, Adamo D

    2017-07-01

    Seed oil and flours have been attracting the interest of researchers and industry, since they contain various bioactive components. We monitored the effects of ripening on lipids, monomeric flavan-3-ols, proanthocyanidins and tocols concentration in seed extracts from organically cultivated cv. Sangiovese vines. Linoleic acid was the most abundant fatty acid, followed by oleic, palmitic and stearic acids. The tocols detected were α-tocopherol, α-tocotrienol and γ-tocotrienol. The proanthocyanidins degree of polymerisation ranged from dimers to dodecamers; moreover, monomeric flavan-3-ols and polymeric proanthocyanidins were detected. Total flavan-3-ols (monomers, oligomers and polymers) concentration in grape seeds decreased during ripening. Fatty acids reached the highest level in post-veraison. The concentration of these compounds varied considerably during ripening. Capric acid has been found for the first time in grape seeds. α-Tocopherol and γ-tocotrienol decreased during ripening, while α-tocotrienol increased. The HPLC analysis with fluorimetric detection, conducted for the first time on cv. Sangiovese, revealed that the concentration of flavan-3-ols monomers, oligomeric proanthocyanidins and polymers greatly changed during ripening. These results suggest that the timing of bunch harvest plays a crucial role in the valorisation of grape seed flour. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  6. Molecular investigation of bacterial communities during the manufacturing and ripening of semi-hard Iranian Liqvan cheese.

    PubMed

    Ramezani, M; Hosseini, S M; Ferrocino, I; Amoozegar, M A; Cocolin, L

    2017-09-01

    Liqvan (or Lighvan) is a traditional Iranian cheese from the East Azerbaijan province of Iran, which is made of raw ewe's milk without the addition of a starter. The grazing pastures, environmental conditions and the ancient regional production methods allocate a distinctive microbial ecology to this type of cheese, and these factors are consequently associated with the quality of the product. In this study, the microbiota of the milk, curd and cheese has been investigated using culture independent approaches. Denaturing gradient gel electrophoresis (DGGE) of the bacteria, 16S rRNA based high-throughput sequencing and enumeration of the live bacterial community by means of quantitative PCR (qPCR) have been used for this purpose. The results showed that the main bacterial population in the milk belonged to both microbial contaminants and lactic acid bacteria (LAB). However, both of these populations were totally replaced by LAB during ripening. The present survey contributes by describing the microbiota of this ancient cheese in more detail during fermentation and ripening. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Ripening and storage conditions of Chétoui and Arbequina olives: Part II. Effect on olive endogenous enzymes and virgin olive oil secoiridoid profile determined by high resolution mass spectrometry.

    PubMed

    Hachicha Hbaieb, Rim; Kotti, Faten; Cortes-Francisco, Nuria; Caixach, Josep; Gargouri, Mohamed; Vichi, Stefania

    2016-11-01

    Several factors affect virgin olive oil (VOO) phenolic profile. The aim of this study was to monitor olive hydrolytic (β-glucosidase) and oxidative (peroxydase, POX, and polyphenoloxydase, PPO) enzymes during olive ripening and storage and to determine their capacity to shape VOO phenolic profile. To this end, olives from the cultivars Chétoui and Arbequina were stored at 4°C or 25°C for 4weeks and their enzymatic activities and oil phenolic profiles were compared to those of ripening olives. We observed different trends in enzymes activities according to cultivar and storage temperature. Secoiridoid compounds, determined by high resolution mass spectrometry (HRMS), and their deacetoxylated, oxygenated, and deacetoxy-oxygenated derivatives were identified and their contents differed between the cultivars according to olive ripening degree and storage conditions. These differences could be due to β-glucosidase, POX and PPO activities changes during olive ripening and storage. Results also show that oxidised phenolic compounds could be a marker of VOO ''freshness". Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Epidermal growth factor receptor inhibitors in cancer treatment: advances, challenges and opportunities.

    PubMed

    Modjtahedi, Helmout; Essapen, Sharadah

    2009-11-01

    Aberrant expression of the epidermal growth factor receptor (EGFR) system has been reported in a wide range of epithelial cancers. In some studies, this has also been associated with a poor prognosis and resistance to the conventional forms of therapies. These discoveries have led to the strategic development of several kinds of EGFR inhibitors, five of which have gained US Food and Drug Administration approval for the treatment of patients with non-small-cell lung cancer (gefitinib and erlotinib), metastatic colorectal cancer (cetuximab and panitumumab), head and neck (cetuximab), pancreatic cancer (erlotinib) and breast (lapatinib) cancer. Despite these advances and recent studies on the predictive value of activating EGFR mutation and KRAS mutations with response in non-small-cell lung cancer and colon cancer patients, there is currently no reliable predictive marker for response to therapy with the anti-EGFR monoclonal antibodies cetuximab and panitumumab or the small molecule EGFR tyrosine kinase inhibitors gefitinib and erlotinib. In particular, there has been no clear association between the expression of EGFR, determined by the US Food and Drug Administration-approved EGFR PharmDX kit, and response to the EGFR inhibitors. Here, we discuss some of the controversial data and explanatory factors as well as future studies for the establishment of more reliable markers for response to therapy with EGFR inhibitors. Such investigations should lead to the selection of a more specific subpopulation of cancer patients who benefit from therapy with EGFR inhibitors, but equally to spare those who will receive no benefit or a detrimental effect from such biological agents.

  9. Gene expression in the pulp of ripening bananas. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products and cDNA cloning of 25 different ripening-related mRNAs.

    PubMed Central

    Medina-Suárez, R; Manning, K; Fletcher, J; Aked, J; Bird, C R; Seymour, G B

    1997-01-01

    mRNA was extracted from the pulp and peel of preclimacteric (d 0) bananas (Musa AAA group, cv Grand Nain) and those exposed to ethylene gas for 24 h and stored in air alone for a further 1 (d 2) and 4 d (d 5). Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products from the pulp and peel of these fruits revealed significant up-regulation of numerous transcripts during ripening. The majority of the changes were initiated by d 2, with the level of these messages increasing during the remainder of the ripening period. Pulp tissue from d 2 was used for the construction of a cDNA library. This library was differentially screened for ripening-related clones using cDNA from d-0 and d-2 pulp by a novel microtiter plate method. In the primary screen 250 up- and down-regulated clones were isolated. Of these, 59 differentially expressed clones were obtained from the secondary screen. All of these cDNAs were partially sequenced and grouped into families after database searches. Twenty-five nonredundant groups of pulp clones were identified. These encoded enzymes were involved in ethylene biosynthesis, respiration, starch metabolism, cell wall degradation, and several other key metabolic events. We describe the analysis of these clones and their possible involvement in ripening. PMID:9342865

  10. Transcriptome analysis during ripening of table grape berry cv. Thompson Seedless

    PubMed Central

    Balic, Iván; Vizoso, Paula; Nilo-Poyanco, Ricardo; Sanhueza, Dayan; Olmedo, Patricio; Sepúlveda, Pablo; Arriagada, Cesar; Defilippi, Bruno G.; Meneses, Claudio

    2018-01-01

    Ripening is one of the key processes associated with the development of major organoleptic characteristics of the fruit. This process has been extensively characterized in climacteric fruit, in contrast with non-climacteric fruit such as grape, where the process is less understood. With the aim of studying changes in gene expression during ripening of non-climacteric fruit, an Illumina based RNA-Seq transcriptome analysis was performed on four developmental stages, between veraison and harvest, on table grapes berries cv Thompson Seedless. Functional analysis showed a transcriptional increase in genes related with degradation processes of chlorophyll, lipids, macromolecules recycling and nucleosomes organization; accompanied by a decrease in genes related with chloroplasts integrity and amino acid synthesis pathways. It was possible to identify several processes described during leaf senescence, particularly close to harvest. Before this point, the results suggest a high transcriptional activity associated with the regulation of gene expression, cytoskeletal organization and cell wall metabolism, which can be related to growth of berries and firmness loss characteristic to this stage of development. This high metabolic activity could be associated with an increase in the transcription of genes related with glycolysis and respiration, unexpected for a non-climacteric fruit ripening. PMID:29320527

  11. Changes in Cuticular Wax Composition of Two Blueberry Cultivars during Fruit Ripening and Postharvest Cold Storage.

    PubMed

    Chu, Wenjing; Gao, Haiyan; Chen, Hangjun; Wu, Weijie; Fang, Xiangjun

    2018-03-21

    Cuticular wax plays an important role for the quality of blueberry fruits. In this study, the cuticular wax composition of two blueberry cultivars, 'Legacy' ( Vaccinium corymbosum) and 'Brightwell' ( Vaccinium ashei), was examined during fruit ripening and postharvest cold storage. The results showed that wax was gradually deposited on the epidermis of blueberry fruits and the content of major wax compounds, except that for diketones, increased significantly during fruit ripening. The total wax content was 2-fold greater in 'Brightwell' blueberries than that in 'Legacy' blueberries during fruit ripening. The total wax content of both cultivars decreased during 30 days of storage at 4 °C, and the variation of cuticular wax composition was cultivar-dependent. The content of diketones decreased significantly in 'Legacy' blueberries, while the content of triterpenoids and aliphatic compounds showed different fold changes in 'Brightwell' blueberries after 30 days of storage at 4 °C. Overall, our study provided a quantitative and qualitative overview of cuticular wax compounds of blueberry fruits during ripening and postharvest cold storage.

  12. Proteome changes in the skin of the grape cultivar Barbera among different stages of ripening

    PubMed Central

    Negri, Alfredo S; Prinsi, Bhakti; Rossoni, Mara; Failla, Osvaldo; Scienza, Attilio; Cocucci, Maurizio; Espen, Luca

    2008-01-01

    Background Grape ripening represents the third phase of the double sigmoidal curve of berry development and is characterized by deep changes in the organoleptic characteristics. In this process, the skin plays a central role in the synthesis of many compounds of interest (e.g. anthocyanins and aroma volatiles) and represents a fundamental protective barrier against damage by physical injuries and pathogen attacks. In order to improve the knowledge on the role of this tissue during ripening, changes in the protein expression in the skin of the red cultivar Barbera at five different stages from véraison to full maturation were studied by performing a comparative 2-DE analysis. Results The proteomic analysis revealed that 80 spots were differentially expressed throughout berry ripening. Applying a two-way hierarchical clustering analysis to these variations, a clear difference between the first two samplings (up to 14 days after véraison) and the following three (from 28 to 49 days after véraison) emerged, thus suggesting that the most relevant changes in protein expression occurred in the first weeks of ripening. By means of LC-ESI-MS/MS analysis, 69 proteins were characterized. Many of these variations were related to proteins involved in responses to stress (38%), glycolysis and gluconeogenesis (13%), C-compounds and carbohydrate metabolism (13%) and amino acid metabolism (10%). Conclusion These results give new insights to the skin proteome evolution during ripening, thus underlining some interesting traits of this tissue. In this view, we observed the ripening-related induction of many enzymes involved in primary metabolism, including those of the last five steps of the glycolytic pathway, which had been described as down-regulated in previous studies performed on whole fruit. Moreover, these data emphasize the relevance of this tissue as a physical barrier exerting an important part in berry protection. In fact, the level of many proteins involved in (a

  13. Effect of immobilized Lactobacillus casei on the evolution of flavor compounds in probiotic dry-fermented sausages during ripening.

    PubMed

    Sidira, Marianthi; Kandylis, Panagiotis; Kanellaki, Maria; Kourkoutas, Yiannis

    2015-02-01

    The effect of immobilized Lactobacillus casei ATCC 393 on wheat grains on the generation of volatile compounds in probiotic dry-fermented sausages during ripening was investigated. For comparison reasons, sausages containing free L. casei cells or no starter culture were also included in the study. Samples were collected after 1, 28 and 45days of ripening and subjected to SPME GC/MS analysis. Both the probiotic culture and the ripening process affected significantly the concentration of all volatile compounds. The significantly highest content of total volatiles, esters, alcohols and miscellaneous compounds was observed in sausages containing the highest amount of immobilized culture (300g/kg of stuffing mixture) ripened for 45days. Principal component analysis of the semi-quantitative data revealed that primarily the concentration of the immobilized probiotic culture affected the volatile composition. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Ripening of pepper (Capsicum annuum) fruit is characterized by an enhancement of protein tyrosine nitration.

    PubMed

    Chaki, Mounira; Álvarez de Morales, Paz; Ruiz, Carmelo; Begara-Morales, Juan C; Barroso, Juan B; Corpas, Francisco J; Palma, José M

    2015-09-01

    Pepper (Capsicum annuum, Solanaceae) fruits are consumed worldwide and are of great economic importance. In most species ripening is characterized by important visual and metabolic changes, the latter including emission of volatile organic compounds associated with respiration, destruction of chlorophylls, synthesis of new pigments (red/yellow carotenoids plus xanthophylls and anthocyanins), formation of pectins and protein synthesis. The involvement of nitric oxide (NO) in fruit ripening has been established, but more work is needed to detail the metabolic networks involving NO and other reactive nitrogen species (RNS) in the process. It has been reported that RNS can mediate post-translational modifications of proteins, which can modulate physiological processes through mechanisms of cellular signalling. This study therefore examined the potential role of NO in nitration of tyrosine during the ripening of California sweet pepper. The NO content of green and red pepper fruit was determined spectrofluorometrically. Fruits at the breaking point between green and red coloration were incubated in the presence of NO for 1 h and then left to ripen for 3 d. Profiles of nitrated proteins were determined using an antibody against nitro-tyrosine (NO2-Tyr), and profiles of nitrosothiols were determined by confocal laser scanning microscopy. Nitrated proteins were identified by 2-D electrophoresis and MALDI-TOF/TOF analysis. Treatment with NO delayed the ripening of fruit. An enhancement of nitrosothiols and nitroproteins was observed in fruit during ripening, and this was reversed by the addition of exogenous NO gas. Six nitrated proteins were identified and were characterized as being involved in redox, protein, carbohydrate and oxidative metabolism, and in glutamate biosynthesis. Catalase was the most abundant nitrated protein found in both green and red fruit. The RNS profile reported here indicates that ripening of pepper fruit is characterized by an enhancement of S

  15. [Massive bleeding symptoms in two patients with factor V inhibitor and antiphospholipid antibodies after treatment with ciprofloxacin].

    PubMed

    Miesbach, Wolfgang; Voigt, Jochen; Peetz, Dirk; Scharrer, Inge

    2003-06-15

    The development of factor V inhibitor is very rare, especially in combination with antiphospholipid antibodies. The paper describes the course of two patients with factor V inhibitor, antiphospholipid antibodies and massive bleeding symptoms after treatment with ciprofloxacin. At that time, ciprofloxacin was the only new drug given. DIAGNOSIS AND CLINICAL COURSE: First changes of the coagulation system were detected 4 days after start of treatment. In one case, occurrence was transient, and normalization was observed after terminating ciprofloxacin treatment. The other case ended with massive muscular and visceral bleedings and cardiovascular failure. Factor V inhibitor and antiphospholipid antibodies could be demonstrated even after termination of ciprofloxacin therapy. The association of treatment with ciprofloxacin and development of factor V inhibitor and antiphospholipid antibodies is probably diagnosed to rarely. These two cases emphasize the necessity of meticulous clarification of a prolonged activated partial thromboplastin time (aPTT) and a drop in prothrombin time (PT) during and after ciprofloxacin treatment.

  16. Bacteriophage biocontrol of Listeria monocytogenes on soft ripened white mold and red-smear cheeses.

    PubMed

    Guenther, Susanne; Loessner, Martin J

    2011-03-01

    Soft-ripened cheeses belong to the type of food most often contaminated with Listeria monocytogenes, and they have been implicated in several outbreaks of listeriosis. Bacteriophages represent an attractive way to combat foodborne pathogens without affecting other properties of the food. We used the broad host range, virulent Listeria phage A511 for control of L. monocytogenes during the production and ripening phases of both types of soft-ripened cheeses, white mold (Camembert-type) cheese, as well as washed-rind cheese with a red-smear surface (Limburger-type). The surfaces of young, unripened cheese were inoculated with 10(1)-10(3) cfu/cm(2)L. monocytogenes strains Scott A (serovar 4b) or CNL 10(3)/2005 (serovar 1/2a). Phage was applied at defined time points thereafter, in single or repeated treatments, at 3 × 10(8) or 1 × 10(9) pfu/cm(2). With Scott A (10(3) cfu/cm(2)) and a single dose of A511 (3 × 10(8) pfu/cm(2)) on camembert-type cheese, viable counts dropped 2.5 logs at the end of the 21 day ripening period. Repeated phage application did not further inhibit the bacteria, whereas a single higher dose (1 × 10(9) pfu/cm(2)) was found to be more effective. On red-smear cheese ripened for 22 days, Listeria counts were down by more than 3 logs. Repeated application of A511 further delayed re-growth of Listeria, but did not affect bacterial counts after 22 days. With lower initial Listeria contamination (10(1)-10(2) cfu/cm(2)), viable counts dropped below the limit of detection, corresponding to more than 6 logs reduction compared to the control. Our data clearly demonstrate the potential of bacteriophage for biocontrol of L. monocytogenes in soft cheese.

  17. PacCYP707A2 negatively regulates cherry fruit ripening while PacCYP707A1 mediates drought tolerance.

    PubMed

    Li, Qian; Chen, Pei; Dai, Shengjie; Sun, Yufei; Yuan, Bing; Kai, Wenbin; Pei, Yuelin; He, Suihuan; Liang, Bin; Zhang, Yushu; Leng, Ping

    2015-07-01

    Sweet cherry is a non-climacteric fruit and its ripening is regulated by abscisic acid (ABA) during fruit development. In this study, four cDNAs (PacCYP707A1-4) encoding 8'-hydroxylase, a key enzyme in the oxidative catabolism of ABA, were identified in sweet cherry fruits using tobacco rattle virus-induced gene silencing (VIGS) and particle bombardment approaches. Quantitative real-time PCR confirmed significant down-regulation of target gene transcripts in VIGS-treated cherry fruits. In PacCYP707A2-RNAi-treated fruits, ripening and fruit colouring were promoted relative to control fruits, and both ABA accumulation and PacNCED1 transcript levels were up-regulated by 140%. Silencing of PacCYP707A2 by VIGS significantly altered the transcripts of both ABA-responsive and ripening-related genes, including the ABA metabolism-associated genes NCED and CYP707A, the anthocyanin synthesis genes PacCHS, PacCHI, PacF3H, PacDFR, PacANS, and PacUFGT, the ethylene biosynthesis gene PacACO1, and the transcription factor PacMYBA. The promoter of PacMYBA responded more strongly to PacCYP707A2-RNAi-treated fruits than to PacCYP707A1-RNAi-treated fruits. By contrast, silencing of PacCYP707A1 stimulated a slight increase in fruit colouring and enhanced resistance to dehydration stress compared with control fruits. These results suggest that PacCYP707A2 is a key regulator of ABA catabolism that functions as a negative regulator of fruit ripening, while PacCYP707A1 regulates ABA content in response to dehydration during fruit development. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  18. Ostwald ripening and interparticle-diffraction effects for illite crystals

    USGS Publications Warehouse

    Eberl, D.D.; Srodon, J.

    1988-01-01

    The Warren-Averbach method, an X-ray diffraction (XRD) method used to measure mean particle thickness and particle-thickness distribution, is used to restudy sericite from the Silverton caldera. Apparent particle-thickness distributions indicate that the clays may have undergone Ostwald ripening and that this process has modified the K-Ar ages of the samples. The mechanism of Ostwald ripening can account for many of the features found for the hydrothermal alteration of illite. Expandabilities measured by the XRD peak-position method for illite/smectites (I/S) from various locations are smaller than expandabilities measured by transmission electron microscopy (TEM) and by the Warren-Averbach (W-A) method. This disparity is interpreted as being related to the presence of nonswelling basal surfaces that form the ends of stacks of illite particles (short-stack effect), stacks that, according to the theory of interparticle diffraction, diffract as coherent X-ray scattering domains. -from Authors

  19. The influence of the wooden equipment employed for cheese manufacture on the characteristics of a traditional stretched cheese during ripening.

    PubMed

    Di Grigoli, Antonino; Francesca, Nicola; Gaglio, Raimondo; Guarrasi, Valeria; Moschetti, Marta; Scatassa, Maria Luisa; Settanni, Luca; Bonanno, Adriana

    2015-04-01

    The influence of the wooden equipment used for the traditional cheese manufacturing from raw milk was evaluated on the variations of chemico-physical characteristics and microbial populations during the ripening of Caciocavallo Palermitano cheese. Milk from two farms (A, extensive; B, intensive) was processed in traditional and standard conditions. Chemical and physical traits of cheeses were affected by the farming system and the cheese making technology, and changed during ripening. Content in NaCl and N soluble was lower, and paste consistency higher in cheese from the extensive farm and traditional technology, whereas ripening increased the N soluble and the paste yellow and consistency. The ripening time decreased the number of all lactic acid bacteria (LAB) groups, except enterococci detected at approximately constant levels (10(4) and 10(5) cfu g(-1) for standard and traditional cheeses, respectively), till 120 d of ripening. In all productions, at each ripening time, the levels detected for enterococci were lower than those for the other LAB groups. The canonical discriminant analysis of chemical, physical and microbiological data was able to separate cheeses from different productions and ripening time. The dominant LAB were isolated, phenotypically characterised and grouped, genetically differentiated at strain level and identified. Ten species of LAB were found and the strains detected at the highest levels were Pediococcus acidilactici and Lactobacillus casei. Ten strains, mainly belonging to Lactobacillus rhamnosus and Lactobacillus fermentum showed an antibacterial activity. The comparison of the polymorphic profiles of the LAB strains isolated from the wooden vat with those of the strains collected during maturation, showed the persistence of three enterococci in traditional cheeses, with Enterococcus faecalis found at dominant levels over the Enterococcus population till 120 d; the absence of these strains in the standard productions evidenced the

  20. Polycomb-group protein SlMSI1 represses the expression of fruit-ripening genes to prolong shelf life in tomato.

    PubMed

    Liu, Dan-Dan; Zhou, Li-Jie; Fang, Mou-Jing; Dong, Qing-Long; An, Xiu-Hong; You, Chun-Xiang; Hao, Yu-Jin

    2016-08-25

    Polycomb-group (PcG) protein MULTICOPY SUPPRESSOR OF IRA1 (MSI1) protein is an evolutionarily conserved developmental suppressor and plays a crucial role in regulating epigenetic modulations. However, the potential role and function of MSI1 in fleshy fruits remain unknown. In this study, SlMSI1 was cloned and transformed into tomato to explore its function. The quantitative real-time PCR results showed that SlMSI1 was highly expressed in flowers and fruits and that its transcript and protein levels were significantly decreased in fruits after the breaker stage. Additionally, SlMSI1-overexpressing transgenic tomatoes displayed abnormal non-ripening fruit formation, whereas its suppression promoted fruit ripening in transgenic tomatoes. Quantitative real-time PCR assays also showed that RIN and its regulons were decreased in SlMSI1 overexpression transgenic tomato fruits. Furthermore, RNA-seq analysis demonstrated that SlMSI1 inhibits fruit ripening by negatively regulating a large set of fruit-ripening genes in addition to RIN and its regulons. Finally, genetic manipulation of SlMSI1 and RIN successfully prolonged the fruit shelf life by regulating the fruit-ripening genes in tomato. Our findings reveal a novel regulatory function of SlMSI1 in fruit ripening and provide a new regulator that may be useful for genetic engineering and modification of fruit shelf life.

  1. Polycomb-group protein SlMSI1 represses the expression of fruit-ripening genes to prolong shelf life in tomato

    PubMed Central

    Liu, Dan-Dan; Zhou, Li-Jie; Fang, Mou-Jing; Dong, Qing-Long; An, Xiu-Hong; You, Chun-Xiang; Hao, Yu-Jin

    2016-01-01

    Polycomb-group (PcG) protein MULTICOPY SUPPRESSOR OF IRA1 (MSI1) protein is an evolutionarily conserved developmental suppressor and plays a crucial role in regulating epigenetic modulations. However, the potential role and function of MSI1 in fleshy fruits remain unknown. In this study, SlMSI1 was cloned and transformed into tomato to explore its function. The quantitative real-time PCR results showed that SlMSI1 was highly expressed in flowers and fruits and that its transcript and protein levels were significantly decreased in fruits after the breaker stage. Additionally, SlMSI1-overexpressing transgenic tomatoes displayed abnormal non-ripening fruit formation, whereas its suppression promoted fruit ripening in transgenic tomatoes. Quantitative real-time PCR assays also showed that RIN and its regulons were decreased in SlMSI1 overexpression transgenic tomato fruits. Furthermore, RNA-seq analysis demonstrated that SlMSI1 inhibits fruit ripening by negatively regulating a large set of fruit-ripening genes in addition to RIN and its regulons. Finally, genetic manipulation of SlMSI1 and RIN successfully prolonged the fruit shelf life by regulating the fruit-ripening genes in tomato. Our findings reveal a novel regulatory function of SlMSI1 in fruit ripening and provide a new regulator that may be useful for genetic engineering and modification of fruit shelf life. PMID:27558543

  2. Fate of Yersinia enterocolitica during manufacture, ripening and storage of Lighvan cheese.

    PubMed

    Hanifian, Shahram; Khani, Sajjad

    2012-05-15

    This study aimed to evaluate the behavior of virulent Yersinia enterocolitica (YE) during the manufacture, ripening and storage of Lighvan cheese with particular reference to strains of YE, initial inoculation level, and storage time. Three strains of YE with low (1 log cfu/ml) and high (3 log cfu/ml) inoculation levels were inoculated to raw whole ewe's milk which was then used for manufacturing of Lighvan cheese. Throughout the manufacturing, ripening and storage periods the number of YE was counted on selective media. Enumerated colonies were then confirmed by duplex PCR using ail and virF genes. Moreover, some microbial and physiochemical characteristics of the cheese samples were examined. According to the results, initial inoculation level and storage time had statistically significant (P<0.01) effects on persistency of YE, while strain type exhibited no statistically significant (P>0.01) impact on survival of the pathogen. Results showed a rapid increase in the number of YE during manufacturing, however, in the ripening and storage periods the number of YE was decreased and eventually it was eliminated in all cheese batches after 4 months of storage. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. SUCROSE NONFERMENTING1-RELATED PROTEIN KINASE2.6, an Ortholog of OPEN STOMATA1, Is a Negative Regulator of Strawberry Fruit Development and Ripening1[OPEN

    PubMed Central

    Dang, Ruihong; Li, Jinxi; Jiang, Jinzhu; Zhang, Ning; Jia, Meiru; Wei, Lingzhi; Li, Ziqiang; Li, Bingbing; Jia, Wensuo

    2015-01-01

    Whereas the regulatory mechanisms that direct fruit ripening have been studied extensively, little is known about the signaling mechanisms underlying this process, especially for nonclimacteric fruits. In this study, we demonstrated that a SUCROSE NONFERMENTING1-RELATED PROTEIN KINASE2, designated as FaSnRK2.6, is a negative regulator of fruit development and ripening in the nonclimacteric fruit strawberry (Fragaria × ananassa) and can also mediate temperature-modulated strawberry fruit ripening. FaSnRK2.6 was identified as an ortholog of OPEN STOMATA1. Levels of FaSnRK2.6 transcript rapidly decreased during strawberry fruit development and ripening. FaSnRK2.6 was found to be capable of physically interacting with strawberry ABSCISIC ACID INSENSITIVE1, a negative regulator in strawberry fruit ripening. RNA interference-induced silencing of FaSnRK2.6 significantly promoted fruit ripening. By contrast, overexpression of FaSnRK2.6 arrested fruit ripening. Strawberry fruit ripening is highly sensitive to temperature, with high temperatures promoting ripening and low temperatures delaying it. As the temperature increased, the level of FaSnRK2.6 expression declined. Furthermore, manipulating the level of FaSnRK2.6 expression altered the expression of a variety of temperature-responsive genes. Taken together, this study demonstrates that FaSnRK2.6 is a negative regulator of strawberry fruit development and ripening and, furthermore, that FaSnRK2.6 mediates temperature-modulated strawberry fruit ripening. PMID:25609556

  4. Effect of Fat Level and the Ripening Time on Quality Traits of Fermented Sausages

    PubMed Central

    Yim, Dong-Gyun; Jang, Kyoung-Hwan; Chung, Ku-Young

    2016-01-01

    The objective of this study was to investigate the effect of the fat reduction on the physicochemical and microbiological characteristics of fermented sausages during ripening and drying. Low fat fermented sausages were produced with different fat levels (30%, 20%, 10%, and 5%) under ripening conditions and fermented process. Samples from each treatment were taken for physicochemical and microbiological analyses on the 0, 1, 2, 3, 4, 5, 7, 10, 14, and 21st day of ripening. In proximate analysis, the fat reduction in sausages produced an increase in moisture, protein and ash contents during ripening and drying (p<0.05). The weight losses were significantly higher in high fat formulations during the first 4 days, whereas those were higher in low fat ones after 10 days of storage (p<0.05). Fat reduction was responsible for an increase in shear force values after 3 days of storage. The volatile basic nitrogen (VBN) value of the low fat samples was significantly higher (p<0.05). Low fat sausages reduced the extent of lipid oxidation. The lower fat level produced redder sausages. Total plate bacteria and Pseudomonas counts of sausages showed no significant differences. Production of low fat sausages resulted in the physicochemical and microbiological attributes equal to or better than the high fat sausages without negative effects, except only a higher VBN and weight loss. PMID:26732335

  5. Predictors of Discontinuation of Tumor Necrosis Factor Inhibitors in Patients with Rheumatoid Arthritis

    PubMed Central

    AGARWAL, SANDEEP K.; GLASS, ROBERTA J.; SHADICK, NANCY A.; COBLYN, JONATHAN S.; ANDERSON, RONALD J.; MAHER, NANCY E.; WEINBLATT, MICHAEL E.; SOLOMON, DANIEL H.

    2009-01-01

    Objective Tumor necrosis factor-α (TNF) inhibitors have transformed management of rheumatoid arthritis (RA); however, many patients discontinue TNF inhibitors. Our goal was to determine the discontinuation rate of TNF inhibitors and identify predictors associated with discontinuation. Methods Enrollees in the Brigham RA Sequential Study (BRASS) formed the eligible cohort. Patients reporting use of a TNF inhibitor with at least 6 months of followup were followed until reporting TNF inhibitor discontinuation or their last study visit if they continued therapy. Potential predictor variables, including demographic and clinical data assessed at baseline and 6 months prior to study endpoint, were identified using a Cox proportional regression. Results Among 961 patients in BRASS, 503 were using a TNF inhibitor with at least 6 months of followup in BRASS (mean length of followup 39 mo, SD 13). Two hundred ten patients (42%) reported discontinuation of TNF inhibitor. Higher physician global scores (hazard ratio 1.27, 95% CI 1.18–1.38) and RA Disease Activity Index scores (HR 1.13, 95% CI 1.05–1.22) 6 months prior to stopping the TNF inhibitor and higher number of TNF inhibitors used previously (HR 1.30, 95% CI 1.03–1.66) were associated with discontinuation of TNF inhibitor. Prior use of synthetic disease modifying antirheumatic drugs (HR 0.50, 95% CI 0.34–0.72) and more years of cumulative methotrexate use (HR 0.24, 95% CI 0.12–0.47) were inversely associated with discontinuation of TNF inhibitor. Conclusion These data demonstrate that a significant number of patients with RA discontinue TNF inhibitors. Several easily characterized clinical variables have a modest predictive association with reduced probability of TNF inhibitor discontinuation. PMID:18634159

  6. [A Jehovah's Witness child with hemophilia B and factor IX inhibitors undergoing scoliosis surgery].

    PubMed

    Chau, Anthony; Wu, John; Ansermino, Mark; Tredwell, Stephen; Purdy, Robert

    2008-01-01

    To describe the successful perioperative hemostatic management of a Jehovah's Witness patient with hemophilia B and anaphylactic inhibitors to factor IX, undergoing scoliosis surgery. A 14 (1/2)-yr-old boy with severe hemophilia B who had a history of anaphylactic inhibitors to factor IX was scheduled to undergo corrective scoliosis surgery. He was initially started on epoetin alfa and iron supplementation to maximize preoperative red cell mass. Additionally, he was placed on a desensitization protocol of recombinant coagulation factor IX (rFIX) and was then treated with activated recombinant coagulation factor VII (rFVIIa) during the postoperative period. Tranexamic acid was given concomitantly. The intraoperative blood loss was approximately 350 mL. The nadir hemoglobin concentration was 111 g.L(-1) on postoperative days one and two. On postoperative day 11, the patient was stable and discharged home with a hemoglobin of 138 g.L(-1). He did not require blood transfusion and no adverse events were observed. The use of rFIX, rFVIIa, erythropoetin, iron, and tranexamic acid before, during and after scoliosis surgery may be a viable and safe option for hemophilia patients with inhibitors, who refuse blood products.

  7. The Microbial Community Dynamics during the Vitex Honey Ripening Process in the Honeycomb

    PubMed Central

    Wen, Yaqin; Wang, Lin; Jin, Yue; Zhang, Jinzhen; Su, Lei; Zhang, Xiaoling; Zhou, Jinhui; Li, Yi

    2017-01-01

    The bacterial and fungal communities of vitex honey were surveyed by sequencing the 16S rRNA gene and the internal transcribed spacer region of ribosomal DNA. Vitex honey samples were analyzed at different stage of ripening; the vitex flower was also analyzed, and the effect of the chemical composition in the experimental setup was assessed. The results confirmed the presence of dominant Bacillus spp. as the dominant bacterial in honey, and yeast related genera was the main fungal in the honey, respectively. Lactococcus and Enterococcus were detected for the first time in honey. The proportion of most of the fungal community decreased during the honey ripening process. Multivariate analyses also showed that the fungal community of 5, 10, and 15 days honey samples tended to cluster together and were completely separated from the 1 day honey sample. The change in the fungal community showed a correlation with the variation in the chemical components, such as moisture and phenolic compounds. Together, these results suggest that ripening of honey could change its microbial composition, and decrease the potential risk of microbiology. PMID:28912763

  8. A Study on the Expression of Genes Involved in Carotenoids and Anthocyanins During Ripening in Fruit Peel of Green, Yellow, and Red Colored Mango Cultivars.

    PubMed

    Karanjalker, G R; Ravishankar, K V; Shivashankara, K S; Dinesh, M R; Roy, T K; Sudhakar Rao, D V

    2018-01-01

    Mango (Mangiferaindica L.) fruits are generally classified based on peel color into green, yellow, and red types. Mango peel turns from green to yellow or red or retain green colors during ripening. The carotenoids and anthocyanins are the important pigments responsible for the colors of fruits. In the present study, peels of different colored cultivars at three ripening stages were characterized for pigments, colors, and gene expression analysis. The yellow colored cultivar "Arka Anmol" showed higher carotenoid content, wherein β-carotene followed by violaxanthin were the major carotenoid compounds that increased during ripening. The red colored cultivars were characterized with higher anthocyanins with cyanidin-3-O-monoglucosides and peonidin-3-O-glucosides as the major anthocyanins. The gene expression analysis by qRT-PCR showed the higher expression of carotenoid biosynthetic genes viz. lycopene-β-cyclase and violaxanthin-de-epoxidase in yellow colored cv. Arka Anmol, and the expression was found to increase during ripening. However, in red colored cv. "Janardhan Pasand," there is increased regulation of all anthocyanin biosynthetic genes including transcription factors MYB and basic helix loop. This indicated the regulation of the anthocyanins by these genes in red mango peel. The results showed that the accumulation pattern of particular pigments and higher expression of specific biosynthetic genes in mango peel impart different colors.

  9. Tissue Specific Localization of Pectin–Ca2+ Cross-Linkages and Pectin Methyl-Esterification during Fruit Ripening in Tomato (Solanum lycopersicum)

    PubMed Central

    Hyodo, Hiromi; Terao, Azusa; Furukawa, Jun; Sakamoto, Naoya; Yurimoto, Hisayoshi; Satoh, Shinobu; Iwai, Hiroaki

    2013-01-01

    Fruit ripening is one of the developmental processes accompanying seed development. The tomato is a well-known model for studying fruit ripening and development, and the disassembly of primary cell walls and the middle lamella, such as through pectin de-methylesterified by pectin methylesterase (PE) and depolymerization by polygalacturonase (PG), is generally accepted to be one of the major changes that occur during ripening. Although many reports of the changes in pectin during tomato fruit ripening are focused on the relation to softening of the pericarp or the Blossom-end rot by calcium (Ca2+) deficiency disorder, the changes in pectin structure and localization in each tissues during tomato fruit ripening is not well known. In this study, to elucidate the tissue-specific role of pectin during fruit development and ripening, we examined gene expression, the enzymatic activities involved in pectin synthesis and depolymerisation in fruit using biochemical and immunohistochemical analyses, and uronic acids and calcium (Ca)-bound pectin were determined by secondary ion-microprobe mass spectrometry. These results show that changes in pectin properties during fruit development and ripening have tissue-specific patterns. In particular, differential control of pectin methyl-esterification occurs in each tissue. Variations in the cell walls of the pericarp are quite different from that of locular tissues. The Ca-binding pectin and hairy pectin in skin cell layers are important for intercellular and tissue–tissue adhesion. Maintenance of the globular form and softening of tomato fruit may be regulated by the arrangement of pectin structures in each tissue. PMID:24236073

  10. Identification and expression analysis of four 14-3-3 genes during fruit ripening in banana (Musa acuminata L. AAA group, cv. Brazilian).

    PubMed

    Li, Mei-Ying; Xu, Bi-Yu; Liu, Ju-Hua; Yang, Xiao-Liang; Zhang, Jian-Bin; Jia, Cai-Hong; Ren, Li-Cheng; Jin, Zhi-Qiang

    2012-02-01

    To investigate the regulation of 14-3-3 proteins in banana (Musa acuminata L. AAA group, cv. Brazilian) fruit postharvest ripening, four cDNAs encoding 14-3-3 proteins were isolated from banana and designated as Ma-14-3-3a, Ma-14-3-3c, Ma-14-3-3e, and Ma-14-3-3i, respectively. Amino acid sequence alignment showed that the four 14-3-3 proteins shared a highly conserved core structure and variable C-terminal as well as N-terminal regions with 14-3-3 proteins from other plant species. Phylogenetic analysis revealed that the four 14-3-3 genes belong to the non-ε groups. They were differentially and specifically expressed in various tissues. Real-time RT-PCR analysis indicated that these four genes function differentially during banana fruit postharvest ripening. Three genes, Ma-14-3-3a, Ma-14-3-3c, and Ma-14-3-3e, were significantly induced by exogenous ethylene treatment. However, gene function differed in naturally ripened fruits. Ethylene could induce Ma-14-3-3c expression during postharvest ripening, but expression patterns of Ma-14-3-3a and Ma-14-3-3e suggest that these two genes appear to be involved in regulating ethylene biosynthesis during fruit ripening. No obvious relationship emerged between Ma-14-3-3i expression in naturally ripened and 1-MCP (1-methylcyclopropene)-treated fruit groups during fruit ripening. These results indicate that the 14-3-3 proteins might be involved in various regulatory processes of banana fruit ripening. Further studies will mainly focus on revealing the detailed biological mechanisms of these four 14-3-3 genes in regulating banana fruit postharvest ripening.

  11. Changes in oxidative stress in transgenic RNAi ACO1 tomato fruit during ripening

    NASA Astrophysics Data System (ADS)

    Eglous, Najat Mohamed; Ali, Zainon Mohd; Hassan, Maizom; Zainal, Zamri

    2013-11-01

    Tomato (Solanum Lycopersicum L.) is the second most cultivated vegetable in the world and widely used as a system for studying the role of ethylene during fruit ripening. Our objective was to study the oxidative stress and antioxidative metabolism during ripening of non transgenic tomato and transgenic line-21 tomato which reduced ethylene. The line-21 of transgenic tomato plants (RNAi ACO1) had lower ethylene production and longer shelf-life more than 32 days as compared to the wild-type fruits which have very short shelf-life. In this study, tomato fruit were divided into five different stages (MG: mature green 5%, B: breaker 25%, T: turning 50%, O: orange75%, RR: red ripe100%). The activity of lipoxygenase (LOX) and lipid peroxidation (MDA) were measured to assess changes in oxidative stress. The LOX activity and MDA content decreased significantly obtaining 2.6-fold and 1.2-fold, respectively, as compared to the wild type fruit. However, superoxide dismutase (SOD) and catalase (CAT) activities were increased to 1.9 and 1.2 folds from the mature green to the fully ripe stage in transgenic tomatoes. Furthermore, the wild type tomato increases 1.3 in SOD and 1.6 in CAT activities. The overall results indicate that the wild type tomato fruit showed a faster rate of ripening, parallel to decline in the rate of enzymatic antioxidative systems as compared to the transgenic line-21 tomato fruit. In addition, the results show that the antioxidant capacity is improved during the ripening process and is accompanied by an increase in the oxidative stress.

  12. Kinetic characterization of factor Xa binding using a quenched fluorescent substrate based on the reactive site of factor Xa inhibitor from Bauhinia ungulata seeds.

    PubMed

    Oliva, M L V; Andrade, S A; Juliano, M A; Sallai, R C; Torquato, R J; Sampaio, M U; Pott, V J; Sampaio, C A M

    2003-07-01

    The specific Kunitz Bauhinia ungulata factor Xa inhibitor (BuXI) and the Bauhinia variegata trypsin inhibitor (BvTI) blocked the activity of trypsin, chymotrypsin, plasmin, plasma kallikrein and factor XIIa, and factor Xa inhibition was achieved only by BuXI (K(i) 14 nM). BuXI and BvTI are highly homologous (70%). The major differences are the methionine residues at BuXI reactive site, which are involved in the inhibition, since the oxidized protein no longer inhibits factor Xa but maintains the trypsin inhibition. Quenched fluorescent substrates based on the reactive site sequence of the inhibitors were synthesized and the kinetic parameters of the hydrolysis were determined using factor Xa and trypsin. The catalytic efficiency k(cat)/K(m) 4.3 x 10(7) M(-1)sec(>-1) for Abz-VMIAALPRTMFIQ-EDDnp (lead peptide) hydrolysis by factor Xa was 10(4)-fold higher than that of Boc-Ile-Glu-Gly-Arg-AMC, widely used as factor Xa substrate. Lengthening of the substrate changed its susceptibility to factor Xa hydrolysis. Both methionine residues in the substrate influence the binding to factor Xa. Serine replacement of threonine (P(1)') decreases the catalytic efficiency by four orders of magnitude. Factor Xa did not hydrolyze the substrate containing the reactive site sequence of BvTI, that inhibits trypsin inhibitor but not factor Xa. Abz-VMIAALPRTMFIQ-EDDnp prolonged both the prothrombin time and the activated partial thromboplastin time, and the other modified substrates used in this experiment altered blood-clotting assays.

  13. Development of Inhibitors Targeting Hypoxia-Inducible Factor 1 and 2 for Cancer Therapy

    PubMed Central

    Yu, Tianchi

    2017-01-01

    Hypoxia is frequently observed in solid tumors and also one of the major obstacles for effective cancer therapies. Cancer cells take advantage of their ability to adapt hypoxia to initiate a special transcriptional program that renders them more aggressive biological behaviors. Hypoxia-inducible factors (HIFs) are the key factors that control hypoxia-inducible pathways by regulating the expression of a vast array of genes involved in cancer progression and treatment resistance. HIFs, mainly HIF-1 and -2, have become potential targets for developing novel cancer therapeutics. This article reviews the updated information in tumor HIF pathways, particularly recent advances in the development of HIF inhibitors. These inhibitors interfere with mRNA expression, protein synthesis, protein degradation and dimerization, DNA binding and transcriptional activity of HIF-1 and -2, or both. Despite efforts in the past two decades, no agents directly inhibiting HIFs have been approved for treating cancer patients. By analyzing results of the published reports, we put the perspectives at the end of the article. The therapeutic efficacy of HIF inhibitors may be improved if more efforts are devoted on developing agents that are able to simultaneously target HIF-1 and -2, increasing the penetrating capacity of HIF inhibitors, and selecting suitable patient subpopulations for clinical trials. PMID:28332352

  14. Control of oxidation-reduction potential during Cheddar cheese ripening and its effect on the production of volatile flavour compounds.

    PubMed

    Caldeo, Veronica; Hannon, John A; Hickey, Dara-Kate; Waldron, Dave; Wilkinson, Martin G; Beresford, Thomas P; McSweeney, Paul L H

    2016-11-01

    In cheese, a negative oxidation-reduction (redox) potential is required for the stability of aroma, especially that associated with volatile sulphur compounds. To control the redox potential during ripening, redox agents were added to the salted curd of Cheddar cheese before pressing. The control cheese contained only salt, while different oxidising or reducing agents were added with the NaCl to the experimental cheeses. KIO3 (at 0·05, 0·1 and 1%, w/w) was used as the oxidising agent while cysteine (at 2%, w/w) and Na2S2O4 (at 0·05 and 0·1%, w/w) were used as reducing agents. During ripening the redox potential of the cheeses made with the reducing agents did not differ significantly from the control cheese (E h ≈ -120 mV) while the cheeses made with 0·1 and 0·05% KIO3 had a significantly higher and positive redox potential in the first month of ripening. Cheese made with 1% KIO3 had positive values of redox potential throughout ripening but no starter lactic acid bacteria survived in this cheese; however, numbers of starter organisms in all other cheeses were similar. Principal component analysis (PCA) of the volatile compounds clearly separated the cheeses made with the reducing agents from cheeses made with the oxidising agents at 2 month of ripening. Cheeses with reducing agents were characterized by the presence of sulphur compounds whereas cheeses made with KIO3 were characterized mainly by aldehydes. At 6 month of ripening, separation by PCA was less evident. These findings support the hypothesis that redox potential could be controlled during ripening and that this parameter has an influence on the development of cheese flavour.

  15. Influence assessment of a lab-scale ripening process on the quality of mechanically-biologically treated MSW for possible recovery.

    PubMed

    Di Lonardo, Maria Chiara; Binner, Erwin; Lombardi, Francesco

    2015-09-01

    In this study, the influence of an additional ripening process on the quality of mechanically-biologically treated MSW was evaluated in the prospective of recovering the end material, rather than landfilling. The biostabilised waste (BSW) coming from one of the MBT plants of Rome was therefore subjected to a ripening process in slightly aerated lab test cells. An in-depth investigation on the biological reactivity was performed by means of different types of tests (aerobic and anaerobic biological tests, as well as FT-IR spectroscopy method). A physical-chemical characterisation of waste samples progressively taken during the ripening phase was carried out, as well. In addition, the ripened BSW quality was assessed by comparing the characteristics of a compost sampled at the composting plant of Rome which treat source segregated organic wastes. Results showed that the additional ripening process allowed to obtain a better quality of the biostabilised waste, by achieving a much higher biological stability compared to BSW as-received and similar to that of the tested compost. An important finding was the lower heavy metals (Co, Cr, Cu, Ni, Pb and Zn) release in water phase at the end of the ripening compared to the as-received BSW, showing that metals were mainly bound to solid organic matter. As a result, the ripened waste, though not usable in agriculture as found for the compost sample, proved anyhow to be potentially suitable for land reclamation purposes, such as in landfills as cover material or mixed with degraded and contaminated soil for organic matter and nutrients supply and for metals recovery, respectively. In conclusion the study highlights the need to extend and optimise the biological treatment in the MBT facilities and opens the possibility to recover the output waste instead of landfilling. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Pectins, Hemicelluloses and Celluloses Show Specific Dynamics in the Internal and External Surfaces of Grape Berry Skin During Ripening.

    PubMed

    Fasoli, Marianna; Dell'Anna, Rossana; Dal Santo, Silvia; Balestrini, Raffaella; Sanson, Andrea; Pezzotti, Mario; Monti, Francesca; Zenoni, Sara

    2016-06-01

    Grapevine berry skin is a complex structure that contributes to the final size and shape of the fruit and affects its quality traits. The organization of cell wall polysaccharides in situ and their modification during ripening are largely uncharacterized. The polymer structure of Corvina berry skin, its evolution during ripening and related modifying genes were determined by combing mid-infrared micro-spectroscopy and multivariate statistical analysis with transcript profiling and immunohistochemistry. Spectra were acquired in situ using a surface-sensitive technique on internal and external sides of the skin without previous sample pre-treatment, allowing comparison of the related cell wall polymer dynamics. The external surface featured cuticle-related bands; the internal surface showed more adsorbed water. Application of surface-specific normalization revealed the major molecular changes related to hemicelluloses and pectins in the internal surface and to cellulose and pectins in the external surface and that they occur between mid-ripening and full ripening in both sides of the skin. Transcript profiling of cell wall-modifying genes indicated a general suppression of cell wall metabolism during ripening. Genes related to pectin metabolism-a β-galactosidase, a pectin(methyl)esterase and a pectate lyase-and a xyloglucan endotransglucosylase/hydrolase, involved in hemicellulose modification, showed enhanced expression. In agreement with Fourier transform infrared spectroscopy, patterns due to pectin methyl esterification provided new insights into the relationship between pectin modifications and the associated transcript profile during skin ripening. This study proposes an original description of polymer dynamics in grape berries during ripening, highlighting differences between the internal and external sides of the skin. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For

  17. SlNCED1 and SlCYP707A2: key genes involved in ABA metabolism during tomato fruit ripening

    PubMed Central

    Ji, Kai; Kai, Wenbin; Zhao, Bo; Sun, Yufei; Yuan, Bing; Dai, Shengjie; Li, Qian; Chen, Pei; Wang, Ya; Pei, Yuelin; Wang, Hongqing; Guo, Yangdong; Leng, Ping

    2014-01-01

    Abscisic acid (ABA) plays an important role in fruit development and ripening. Here, three NCED genes encoding 9-cis-epoxycarotenoid dioxygenase (NCED, a key enzyme in the ABA biosynthetic pathway) and three CYP707A genes encoding ABA 8′-hydroxylase (a key enzyme in the oxidative catabolism of ABA) were identified in tomato fruit by tobacco rattle virus-induced gene silencing (VIGS). Quantitative real-time PCR showed that VIGS-treated tomato fruits had significant reductions in target gene transcripts. In SlNCED1-RNAi-treated fruits, ripening slowed down, and the entire fruit turned to orange instead of red as in the control. In comparison, the downregulation of SlCYP707A2 expression in SlCYP707A2-silenced fruit could promote ripening; for example, colouring was quicker than in the control. Silencing SlNCED2/3 or SlCYP707A1/3 made no significant difference to fruit ripening comparing RNAi-treated fruits with control fruits. ABA accumulation and SlNCED1transcript levels in the SlNCED1-RNAi-treated fruit were downregulated to 21% and 19% of those in control fruit, respectively, but upregulated in SlCYP707A2-RNAi-treated fruit. Silencing SlNCED1 or SlCYP707A2 by VIGS significantly altered the transcripts of a set of both ABA-responsive and ripening-related genes, including ABA-signalling genes (PYL1, PP2C1, and SnRK2.2), lycopene-synthesis genes (SlBcyc, SlPSY1 and SlPDS), and cell wall-degrading genes (SlPG1, SlEXP, and SlXET) during ripening. These data indicate that SlNCED1 and SlCYP707A2 are key genes in the regulation of ABA synthesis and catabolism, and are involved in fruit ripening as positive and negative regulators, respectively. PMID:25039074

  18. Quantitative Analysis of Milk-Derived microRNAs and Microbiota during the Manufacturing and Ripening of Soft Cheese.

    PubMed

    Oh, Sangnam; Park, Mi-Ri; Ryu, Sangdon; Maburutse, Brighton; Kim, Ji-Uk; Kim, Younghoon

    2017-09-28

    MicroRNAs (miRNAs) are abundant in bovine milk and milk derived from other livestock, and they have functional roles in infants and in the secretion process of mammary glands. However, few studies have evaluated miRNAs in dairy processes, such as during cheese making and ripening. Thus, we investigated the characteristics of milk-derived miRNAs during the manufacturing and ripening of Camembert cheese as well as the microbiota present using the quantitative reverse transcription polymer chain reaction (RT-qPCR) and 16S rRNA pyrosequencing, respectively. Pyrosequencing showed that the cheese microbiota changed dramatically during cheese processing, including during the pasteurization, starter culture, and ripening stages. Our results indicated that the RNA contents per 200 mg/200 μl of the sample increased significantly during cheese-making and ripening. The inner cheese fractions had higher RNA contents than the surfaces after 12 and 22 days of ripening in a timedependent manner (21.9 and 13.2 times higher in the inner and surface fractions than raw milk, respectively). We performed a comparative analysis of the miRNAs in each fraction by RT-qPCR. Large amounts of miRNAs ( miR-93, miR-106a, miR-130, miR-155, miR-181a , and miR- 223 ) correlated with immune responses and mammary glands were present in aged cheese, with the exception of miR-223 , which was not present on the surface. Considerable amounts of miRNAs were also detected in whey, which is usually disposed of during the cheese-making process. Unexpectedly, there were no significant correlations between immune-related miRNAs and the microbial populations during cheese processing. Taken together, these results show that various functional miRNAs are present in cheese during its manufacture and that they are dramatically increased in amount in ripened Camembert cheese, with differences according to depth.

  19. Correlation between ethylene emission and skin colour changes during papaya (Carica papaya L.) fruit ripening

    NASA Astrophysics Data System (ADS)

    da Silva, M. G.; Oliveira, J. G.; Vitoria, A. P.; Corrêa, S. F.; Pereira, M. G.; Campostrini, E.; Santos, E. O.; Cavalli, A.; Vargas, H.

    2005-06-01

    The skin colour changes and ethylene emission rates were monitored during papaya (C. papaya L.) fruit ripening. Two groups of papaya (‘Formosa’ and ‘Solo’) were applied in this study. The total colour difference was used as measured parameter and the corresponding half time of its saturation was used as correlation parameter. A high correlation factor between the saturation half time and corresponding climacteric peak time was found. It was concluded that high ethylene emission rate in ‘Solo’ fruit promotes a quick change of the total colour difference.

  20. Microbial Ecology Dynamics Reveal a Succession in the Core Microbiota Involved in the Ripening of Pasta Filata Caciocavallo Pugliese Cheese

    PubMed Central

    De Pasquale, Ilaria; Buchin, Solange; De Angelis, Maria; Gobbetti, Marco

    2014-01-01

    seemed to be the main factor responsible for cheese ripening. PMID:25085486

  1. Ripening of Pithecellobium dulce (Roxb.) Benth. [Guamúchil] Fruit: Physicochemical, Chemical and Antioxidant Changes.

    PubMed

    Wall-Medrano, Abraham; González-Aguilar, Gustavo A; Loarca-Piña, Guadalupe F; López-Díaz, José A; Villegas-Ochoa, Mónica A; Tortoledo-Ortiz, Orlando; Olivas-Aguirre, Francisco J; Ramos-Jiménez, Arnulfo; Robles-Zepeda, Ramón

    2016-12-01

    The fruit of Guamúchil is an excellent source of bioactive compounds for human health although their natural occurrence could be affected by the ripening process. The aim was to evaluate some physicochemical, chemical and antioxidant changes in guamúchil fruit during six ripening stages (I to VI). A defined trend (p ≤ 0.003) was observed for color [°Hue, 109 (light green) to 20 (dark red)], anthocyanins (+571 %), soluble solids (+0.33 o Brix), ash (+16 %), sucrose (-91 %), proanthocyanidins (63 %), ascorbic acid (-52 %) and hydrolysable PC (-21 %). Carotenoids were not detected and chlorogenic acid was the most abundant phenolic compound. Maximal availability of these bioactives per ripening stage (p ≤ 0.03) was as follows: I (protein/ lipids/ sucrose/ proanthocyanidins/ hydrolysable phenolics), II (total sugars/ascorbic acid), III (total phenolics), IV (flavonoids/ chlorogenic acid) and VI (fructose/ glucose/ anthocyanins). Color change was explained by sucrose (β = 0.47) and anthocyanin (β = 0.20) contents (p < 0.001). Radical scavenging capacity (ORAC, DPPH and TEAC) strongly correlated with total PC (r = 0.49-0.65, p ≤ 0.001) but 89 % of ORAC's associated variance was explained by anthocyanin + sucrose + ascorbic acid (p ≤ 0.0001). Guamúchil fruit could be a more convenient source of specific bioactive compounds if harvested at different ripening stages.

  2. Use of a Cyclooxygenase-2 Inhibitor Does Not Inhibit Platelet Activation or Growth Factor Release From Platelet-Rich Plasma.

    PubMed

    Ludwig, Hilary C; Birdwhistell, Kate E; Brainard, Benjamin M; Franklin, Samuel P

    2017-12-01

    It remains unestablished whether use of cyclooxygenase (COX)-2 inhibitors impairs platelet activation and anabolic growth factor release from platelets in platelet-rich plasma (PRP). The purpose of this study was to assess the effects of a COX-2 inhibitor on platelet activation and anabolic growth factor release from canine PRP when using a clinically applicable PRP activator and to determine whether a 3-day washout would be sufficient to abrogate any COX-2 inhibitor-related impairment on platelet function. Controlled laboratory study. Ten healthy dogs underwent blood collection and PRP preparation. Dogs were then administered a COX-2 inhibitor for 7 days, after which PRP preparation was repeated. The COX-2 inhibitor was continued for 4 more days and PRP preparation performed a third time, 3 days after discontinuation of the COX-2 inhibitor. Immediately after PRP preparation, the PRP was divided into 4 aliquots: 2 unactivated and 2 activated using human γ-thrombin (HGT). One activated and 1 unactivated sample were assessed using flow cytometry for platelet expression of CD62P and platelet-bound fibrinogen using the canine activated platelet-1 (CAP1) antibody. The 2 remaining samples were centrifuged and the supernatant assayed for transforming growth factor-β1 (TGF-β1), platelet-derived growth factor-BB (PDGF-BB), and thromboxane B2 (TXB2) concentrations. Differences in platelet activation and TGF-β1, PDGF-BB, and TXB2 concentrations over the 3 study weeks were evaluated using a 1-way repeated-measures ANOVA, and comparisons between activated and unactivated samples within a study week were assessed with paired t tests. There were no statistically significant ( P > .05) effects of the COX-2 inhibitor on percentage of platelets positive for CD62P or CAP1 or on concentrations of TGF-β1, PDGF-BB, or TXB2. All unactivated samples had low levels of activation or growth factor concentrations and significantly ( P < .05) greater activation and growth factor

  3. Evolution of the taste of a bitter Camembert cheese during ripening: characterization of a matrix effect.

    PubMed

    Engel, E; Nicklaus, S; Septier, C; Salles, C; Le Quéré, J L

    2001-06-01

    The objective of this study was to characterize the effect of ripening on the taste of a typically bitter Camembert cheese. The first step was to select a typically bitter cheese among several products obtained by different processes supposed to enhance this taste defect. Second, the evolution of cheese taste during ripening was characterized from a sensory point of view. Finally, the relative impact of fat, proteins, and water-soluble molecules on cheese taste was determined by using omission tests performed on a reconstituted cheese. These omission tests showed that cheese taste resulted mainly from the gustatory properties of water-soluble molecules but was modulated by a matrix effect due to fat, proteins, and cheese structure. The evolution of this matrix effect during ripening was discussed for each taste characteristic.

  4. Pyroglutamic acid in cheese: presence, origin, and correlation with ripening time of Grana Padano cheese.

    PubMed

    Mucchetti, G; Locci, F; Gatti, M; Neviani, E; Addeo, F; Dossena, A; Marchelli, R

    2000-04-01

    Pyroglutamic acid is present in many cheese varieties and particularly in high amounts (0.5 g/100 g of cheese) in extensively ripened Italian cheeses (Grana Padano and Parmigiano Reggiano) that are produced with thermophilic lactic acid bacteria as starters. The mechanism of pyroglutamic acid formation in cheese seems to be mostly enzymatic, as demonstrated by the presence of only L-pyroglutamic acid enantiomer. Thermophilic lactobacilli are involved in pyroglutamic acid production, as suggested by the low pyroglutamic acid content found in Bagos, a ripened Italian mountain cheese produced without addition of starter. Because milk pasteurization did not influence the pyroglutamic acid content in the ripened Grana Padano cheese, the formation of pyroglutamic acid mainly depends on the whey starter microflora rather than that of raw milk. Pyroglutamic acid concentration is linearly correlated (R2 = 0.94) with the age of Grana Padano cheese.

  5. Specific metabolic activity of ripening bacteria quantified by real-time reverse transcription PCR throughout Emmental cheese manufacture.

    PubMed

    Falentin, Hélène; Postollec, Florence; Parayre, Sandrine; Henaff, Nadine; Le Bivic, Pierre; Richoux, Romain; Thierry, Anne; Sohier, Danièle

    2010-11-15

    Bacterial communities of fermented foods are usually investigated by culture-dependent methods. Real-time quantitative PCR (qPCR) and reverse transcription (RT)-qPCR offer new possibilities to quantify the populations present and their metabolic activity. The aim of this work was to develop qPCR and RT-qPCR methods to assess the metabolic activity and the stress level of the two species used as ripening cultures in Emmental cheese manufacture, Propionibacterium freudenreichii and Lactobacillus paracasei. Three small scale (1/100) microbiologically controlled Emmental cheeses batches were manufactured and inoculated with Lactobacillus helveticus, Streptococcus thermophilus, P. freudenreichii and L. paracasei. At 12 steps of cheese manufacture and ripening, the populations of P. freudenreichii and L. paracasei were quantified by numerations on agar media and by qPCR. 16S, tuf and groL transcript levels were quantified by RT-qPCR. Sampling was carried out in triplicate. qPCR and RT-qPCR assessments were specific, efficient and linear. The quantification limit was 10(3) copies of cells or cDNA/g of cheese. Cell quantifications obtained by qPCR gave similar results than plate count for P. freudenreichii growth and 0.5 to 1 log lower in the stationary phase. Bacterial counts and qPCR quantifications showed that L. paracasei began to grow during the pressing step while P. freudenreichii began to grow from the beginning of ripening (in the cold room). Tuf cDNA quantification results suggested that metabolic activity of L. paracasei reached a maximum during the first part of the ripening (in cold room) and decreased progressively during ripening (in the warm room). Metabolic activity of P. freudenreichii was maximum at the end of cold ripening room and was stable during the first two weeks in warm room. After lactate exhaustion (after two weeks of warm room), the number of tuf cDNA decreased reflecting reduced metabolic activity. For L. paracasei, groL cDNA were stable

  6. Recombinant factor VIIa (eptacog alfa): a pharmacoeconomic review of its use in haemophilia in patients with inhibitors to clotting factors VIII or IX.

    PubMed

    Lyseng-Williamson, Katherine A; Plosker, Greg L

    2007-01-01

    Recombinant factor VIIa (NovoSeven; also known as recombinant activated factor VII or eptacog alfa) is indicated as an intravenous haemostatic agent in haemophilia patients with inhibitors to clotting factors VIII or IX. In noncomparative trials in haemophilia patients with inhibitors, on-demand home treatment with recombinant factor VIIa was effective in controlling episodes of mild to moderate bleeding and well tolerated, with early treatment being associated with a greater rate of success and the need for fewer doses than delayed treatment. Prophylactic treatment with recombinant factor VIIa was also effective in maintaining haemostasis in patients with this indication undergoing surgery. Relative to prior treatment with plasma-derived agents, treatment with recombinant factor VIIa was associated with improvements in health-related quality of life in a cost-utility study in haemophilia patients with inhibitors in Australia. In well designed decision-model cost analyses conducted from a healthcare payer perspective in several countries, on-demand treatment with recombinant factor VIIa to control mild to moderate bleeding episodes in this patient population was predicted to be cost saving or cost neutral relative to on-demand treatment with intravenous activated prothrombin complex concentrate (aPCC). Although the acquisition cost of recombinant factor VIIa was greater than that of aPCC in some studies, the greater initial efficacy of recombinant factor VIIa than aPCC resulted in lower predicted total medical costs. Results were generally robust to plausible changes in key parameters. Orthopaedic surgery with recombinant factor VIIa to maintain haemostasis in haemophilia patients with inhibitors was generally predicted to be cost saving, relative to not having surgery, over the medium to long term in modelled cost analyses from a healthcare payer perspective in the UK and US. The initial cost of surgery was high, but the difference in costs between patients

  7. Crystal Structure of a Two-domain Fragment of Hepatocyte Growth Factor Activator Inhibitor-1

    PubMed Central

    Hong, Zebin; De Meulemeester, Laura; Jacobi, Annemarie; Pedersen, Jan Skov; Morth, J. Preben; Andreasen, Peter A.; Jensen, Jan K.

    2016-01-01

    Hepatocyte growth factor activator inhibitor-1 (HAI-1) is a type I transmembrane protein and inhibitor of several serine proteases, including hepatocyte growth factor activator and matriptase. The protein is essential for development as knock-out mice die in utero due to placental defects caused by misregulated extracellular proteolysis. HAI-1 contains two Kunitz-type inhibitor domains (Kunitz), which are generally thought of as a functionally self-contained protease inhibitor unit. This is not the case for HAI-1, where our results reveal how interdomain interactions have evolved to stimulate the inhibitory activity of an integrated Kunitz. Here we present an x-ray crystal structure of an HAI-1 fragment covering the internal domain and Kunitz-1. The structure reveals not only that the previously uncharacterized internal domain is a member of the polycystic kidney disease domain family but also how the two domains engage in interdomain interactions. Supported by solution small angle x-ray scattering and a combination of site-directed mutagenesis and functional assays, we show that interdomain interactions not only stabilize the fold of the internal domain but also stimulate the inhibitory activity of Kunitz-1. By completing our structural characterization of the previously unknown N-terminal region of HAI-1, we provide new insight into the interplay between tertiary structure and the inhibitory activity of a multidomain protease inhibitor. We propose a previously unseen mechanism by which the association of an auxiliary domain stimulates the inhibitory activity of a Kunitz-type inhibitor (i.e. the first structure of an intramolecular interaction between a Kunitz and another domain). PMID:27189939

  8. Prevalence of the intron 22 inversion of the factor VIII gene and inhibitor development in Polish patients with severe hemophilia A.

    PubMed

    Sawecka, Jadwiga; Skulimowska, Joanna; Windyga, Jerzy; Lopaciuk, Stanisław; Kościelak, Jerzy

    2005-01-01

    Patients with severe hemophilia A often develop inhibitors (antibodies) against transfused factor VIII. One hundred thirteen Polish patients with severe hemophilia A, who had been treated on demand with cryoprecipitate until 1992 and exclusively with factor VIII concentrates after 1995, were examined for intron 22 inversion by Southern blotting and the presence and magnitude of inhibitor activity in blood as determined by the Bethesda assay. The patients' ages ranged 4--67 years (mean: 33.7+/-12.4 years, median: 32 years). The number of patients with the inversion amounted to 57, while in 56 patients the mutation types were unknown; 47 patients had a distal and 10 patients a proximal type of inversion. Thirteen patients with inversions (22.8%) were found to have inhibitor in their blood. Most patients (14 out of 15) who developed inhibitors in the course of cryoprecipitate therapy were high responders. Conversely, 4 of 5 patients treated between 1992 and 1995 with both cryoprecipitate and intermediate-purity factor VIII concentrates were low responders. One multitransfused patient who had remained inhibitor-free on cryoprecipitate therapy developed inhibitor after receiving a large dose of factor VIII concentrate during surgery. None of these 5 patients developed inhibitors during their 12--40 years of treatment with cryoprecipitate, suggesting that it was less immunogenic than factor VIII concentrates. The prevalence of the intron 22 inversion mutation of the factor VIII gene in Polish hemophiliacs is similar to that in other European countries. Treatment regimens with either cryoprecipitate or virus-inactivated plasma-derived factor VIII concentrates may affect inhibitor formation in hemophilia A patients.

  9. Extension of Ostwald Ripening Theory

    NASA Technical Reports Server (NTRS)

    Baird, J.; Naumann, R.

    1985-01-01

    The objective is to develop models based on the mean field approximation of Ostwald ripening to describe the growth of second phase droplets or crystallites. The models will include time variations in nucleation rate, control of saturation through addition of solute, precipitating agents, changes in temperature, and various surface kinetic effects. Numerical integration schemes have been developed and tested against the asymptotic solution of Liftshitz, Slyozov and Wagner (LSW). A second attractor (in addition to the LSW distribution) has been found and, contrary to the LSW theory, the final distribution is dependent on the initial distribution. A series of microgravity experiments is being planned to test this and other results from this work.

  10. Discovery of novel aminobenzisoxazole derivatives as orally available factor IXa inhibitors

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sakurada, Isao; Endo, Toshiya; Hikita, Katsuyoshi

    2017-06-01

    Using structure based drug design, novel aminobenzisoxazoles as coagulation factor IXa inhibitors were designed and synthesized. Highly selective inhibition of FIXa over FXa was demonstrated. Anticoagulation profile of selected compounds was evaluated by aPTT and PT tests. In vitro ADMET and pharmacokinetic (PK) profiles were also evaluated.

  11. Changing patterns of tumor necrosis factor inhibitor use in 9074 patients with rheumatoid arthritis.

    PubMed

    Yazici, Yusuf; Krasnokutsky, Svetlana; Barnes, Jaime P; Hines, Patricia L; Wang, Jason; Rosenblatt, Lisa

    2009-05-01

    Patients with rheumatoid arthritis (RA) commonly switch between tumor necrosis factor (TNF) inhibitors after failing to control disease activity. Much of the clinical data that support switching to a second TNF agent when one agent fails to work has come from small, short-term studies. We utilized a US insurance claims database to determine patterns of use such as dose escalation, time to discontinuation, and switching between TNF inhibitors in patients with RA. A retrospective analysis was performed using an insurance claims database in the US from 2000 to 2005. TNF inhibitor use, time to switch, dose escalation, and continuation times were analyzed in patients with RA. Nine thousand seventy-four patients with RA started TNF inhibitors during the period 2000 to 2005. Etanercept was the most commonly used TNF inhibitor; infliximab had the highest duration of continuation, about 50% at 2 years. In addition, infliximab showed higher rates of dose escalation compared to etanercept and adalimumab. For all TNF inhibitors, time to switching decreased from 2000 to 2005. TNF inhibitor use patterns changed from 2000 to 2005, with more frequent changes among the different TNF inhibitors and a shorter duration of treatment before the change. Only about 50% of TNF inhibitors are still continued at 2 years, reflecting the difference between randomized clinical trials and real-world experience.

  12. Identification of volatile compound markers during the ripening and senescence of lulo (Solanum quitoense Lam.).

    PubMed

    Corpas Iguarán, Eduardo; Taborda Ocampo, Gonzalo; Tapasco Alzate, Omar

    2018-01-01

    Lulo ( Solanum quitoense Lam.) is an exotic fruit cultivated in Colombia. During ripening and senescence, this climactic fruit undergoes biochemical processes that produce the volatiles responsible for its aroma. This study aimed to evaluate the changes in the volatile content during the ripening and senescence of lulo. Analysis of the volatile composition of lulo harvested in each of its five ripening stages and during its senescence time when stored at 18 ± 2 °C was performed using HS-SPME with GC-MS. Throughout ripening, the most notable change was the transformation of alcohols such as (Z)-3-hexen-1-ol and 1-penten-3-ol to afford esters such as (Z)-3-hexenyl acetate and ketones such as 1-penten-3-one. Some acids reacted with alcohols to produce acetate and hexanoate esters, concentrations which increased more than sixfold between stage one and five. Moreover, all the major compounds were C 6 straight chain compounds related to the lipoxygenase pathway. During senescence, majority of compounds were methyl esters, which increased in concentration consistently until day eight. Remarkably, the content of methyl butanoate increased from 0.9% of the total amount of volatiles on day two up to 76.4% on day eight. Some of these volatiles are probably contributors to the "off flavor" during senescence.

  13. Compositional and biochemical changes in Genestoso cheese, a Spanish raw cow's milk variety, during ripening.

    PubMed

    Arenas, Ricardo; González, Leticia; Sacristán, Noelia; Tornadijo, María E; Fresno, José M

    2015-03-15

    Physicochemical characteristics, proteolysis and lipolysis were studied throughout the ripening of eight batches of a traditional Spanish variety made from raw cow's milk, in order to establish a basis for its industrial production. The main compositional characteristics of this cheese after 60 days of ripening were its high proportion of total solids (TS; 752 g kg⁻¹ of cheese), an average content of protein (452.8 g kg⁻¹ TS) and fat (475.1 g kg⁻¹ TS) and the presence of residual lactose (12.5 g kg⁻¹ TS). Its pH value (4.04) was extremely low. Phosphorus (5.13 g kg⁻¹ TS) and sodium (8.29 g kg⁻¹ TS) were the most abundant mineral elements in cheese, whereas calcium levels (1.92 g kg⁻¹ TS) were very low. Proteolysis extension and depth were very low, which resulted in almost zero degradation of αs1- and β-casein. Fat acidity increased during ripening, reaching final values of 50.1 mg KOH kg⁻¹ of fat. The main free fatty acid was C16:0, followed by C18:1 and C14:0. These results suggest that this variety undergoes a limited proteolysis and moderate lipolysis during ripening. The low pH, low calcium content and limited proteolysis led to a crumbly texture in this cheese variety. © 2014 Society of Chemical Industry.

  14. S-nitrosoglutathione reductase (GSNOR) activity is down-regulated during pepper (Capsicum annuum L.) fruit ripening.

    PubMed

    Rodríguez-Ruiz, Marta; Mioto, Paulo; Palma, José M; Corpas, Francisco J

    2017-08-01

    Pepper (Capsicum annuum L.) is an annual plant species of great agronomic importance whose fruits undergo major metabolic changes through development and ripening. These changes include emission of volatile organic compounds associated with respiration, destruction of chlorophylls and synthesis of new pigments (red/yellow carotenoids plus xanthophylls and anthocyans) responsible for color shift, protein degradation/synthesis and changes in total soluble reducing equivalents. Previous data have shown that, during the ripening of pepper fruit, an enhancement of protein tyrosine nitration takes place. On the other hand, it is well known that S-nitrosoglutathione reductase (GSNOR) activity can modulate the transnitrosylation equilibrium between GSNO and S-nitrosylated proteins and, consequently, regulate cellular NO homeostasis. In this study, GSNOR activity, protein content and gene expression were analyzed in green and red pepper fruits. The content of S-nitrosylated proteins on diaminofluorescein (DAF) gels was also studied. The data show that, while GSNOR activity and protein expression diminished during fruit ripening, S-nitrosylated protein content increased. Some of the protein candidates for S-nitrosylation identified, such as cytochorme c oxidase and peroxiredoxin II E, have previously been described as targets of this posttranslational modification in other plant species. These findings corroborate the important role played by GSNOR activity in the NO metabolism during the process of pepper fruit ripening. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Alterations in the morphological, sugar composition, and volatile flavor properties of petai (Parkia speciosa Hassk.) seed during ripening.

    PubMed

    Asikin, Yonathan; Kusumiyati; Taira, Eizo; Wada, Koji

    2018-04-01

    Petai seeds are one of the well-known strong-smelling foods of the Southeast Asian region that have been harvested and commercially offered in different ripening forms. The current study focused on alterations in the size, color, sugar composition, and volatile flavor properties of petai seeds in the four ripening stages (unripe, mid ripe, ripe, and over-ripe). The ripening process was mainly indicated by the increase in size and weight as seed color turned paler and less greenish. The total sugar content gradually increased during ripening, and then elevated from 1.60 g/100 g (ripe seed) to the level of 2.82 g/100 g in the over-ripe seed. Ripening also altered the volatile flavor composition of petai seed, wherein the predominant aldehydes (hexanal and acetaldehyde) were decreased, and the sulfuric compounds (hydrogen sulfide, methanethiol, and 1,2,4-trithiolane) tended to increase. Additionally, gas chromatography-olfactometry (GC-O) analysis revealed alterations in the perceived odor strength and sensation of each volatile compound and demonstrated volatile flavor profiles, viz. detection percentages of volatile group odor strengths and descriptive odors, of petai seed. These results provide valuable information for monitoring alterations in the physical appearance, sugar composition, and aroma that represent the flavor quality in seasonal petai seed. Copyright © 2018 Elsevier Ltd. All rights reserved.

  16. Impact of casing damaging on aflatoxin B1 concentration during the ripening of dry-fermented meat sausages.

    PubMed

    Pleadin, Jelka; Kovačević, Dragan; Perković, Irena

    2015-01-01

    The aim of this article is to investigate the impact of casing damaging on the formation of aflatoxin B1 (AFB1) during the ripening of dry-fermented meat sausages. The level of AFB1 contamination was determined in 24 samples using the ELISA immunoassay throughout a six-month production period. While with intact casing samples no contamination was observed throughout the whole production process, in damaged casing samples AFB1 was detected in the ripening end-stages in the range of 1.62-4.49 μg/kg. The results showed that casing damaging occurring during long-term ripening of dry-fermented sausages can cause AFB1 contamination, possibly arising on the grounds of diffusion of this mycotoxin from the product surface to its interior.

  17. The home as an appropriate setting for women undertaking cervical ripening before the induction of labour.

    PubMed

    Reid, Margaret; Lorimer, Karen; Norman, Jane E; Bollapragada, Shrikant S; Norrie, John

    2011-02-01

    to explore women's experiences of cervical ripening using isosorbide mononitrate (IMN) in the home as part of the main randomised controlled trial. qualitative study with semi-structured interviews carried out at three weeks post partum. Interview transcripts were analysed to identify recurrent themes, focusing on why women became involved in the study, their views about both the self-medication and the home setting, and whether they would repeat the experience. the home. twenty women enrolled in the main randomised controlled trial. the study is part of a double-blind randomised controlled trial with 350 patients investigating whether a nitric oxide donor (IMN) used in cervical ripening improves the process of induction of labour. women liked the opportunity to remain at home during the cervical ripening process. Timing and setting were central issues; women hoped that it would hasten labour, while the home was seen as a setting offering freedom, security and reassurance, as opposed to the hospital, seen as constraining. Two women reported problems with IMN but the remainder reported that they would repeat the experience. women were very positive about the opportunity to undertake cervical ripening at home. It is important to explore this setting further for appropriate interventions. Copyright © 2009 Elsevier Ltd. All rights reserved.

  18. Induction of AGAMOUS gene expression plays a key role in ripening of tomato sepals in vitro.

    PubMed

    Ishida, B K; Jenkins, S M; Say, B

    1998-03-01

    In vitro culture of VFNT Cherry tomato sepals (calyx) at 16-21 degrees C results in developmental changes that are similar to those that occur in fruit tissue [10]. Sepals become swollen, red, and succulent, produce ethylene, and have increased levels of polygalacturonase RNA. They also produce many flavor volatiles characteristic of ripe tomato fruit and undergo similar changes in sugar content [11]. We examined the expression of the tomato AGAMOUS gene, TAG1, in ripening, in vitro sepal cultures and other tissues from the plant and found that TAG1 RNA accumulates to higher levels than expected from data from other plants. Contrary to reports on the absence of AGAMOUS in sepals, TAG1 RNA levels in green sepals from greenhouse-grown plants is detectable, its concentration increasing with in vitro ripening to levels that were even higher than in red, ripe fruit. Sepals of fruit on transgenic tomato plants that expressed TAG1 ectopically were induced by low temperature to ripen in vivo, producing lycopene and undergoing cell wall softening as is characteristic of pericarpic tissue. We therefore propose that the induction of elevated TAG1 gene expression plays a key role in developmental changes that result in sepal ripening.

  19. Synthesis of CdSe/ZnS and CdTe/ZnS Quantum Dots: Refined Digestive Ripening

    DOE PAGES

    Cingarapu, Sreeram; Yang, Zhiqiang; Sorensen, Christopher M.; ...

    2012-01-01

    We report synthesis of CdSe and CdTe quantum dots (QDs) from the bulk CdSe and CdTe material by evaporation/co-condensation using the solvated metal atom dispersion (SMAD) technique and refined digestive ripening. The outcomes of this new process are (1) the reduction of digestive ripening time by employing ligands (trioctylphosphine oxide (TOPO) and oleylamine (OA)) as capping agent as well as digestive ripening solvent, (2) ability to tune the photoluminescence (PL) from 410 nm to 670 nm, (3) demonstrate the ability of SMAD synthesis technique for other semiconductors (CdTe), (4) direct comparison of CdSe QDs growth with CdTe QDs growth based on digestivemore » ripening times, and (5) enhanced PL quantum yield (QY) of CdSe QDs and CdTe QDs upon covering with a ZnS shell. Further, the merit of this synthesis is the use of bulk CdSe and CdTe as the starting materials, which avoids usage of toxic organometallic compounds, eliminates the hot injection procedure, and size selective precipitation processes. It also allows the possibility of scale up. These QDs were characterized by UV-vis, photoluminescence (PL), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), X-ray photoelectron spectroscopy (XPS), and powder XRD.« less

  20. Effect of elicitors on the evolution of grape phenolic compounds during the ripening period.

    PubMed

    Gómez-Plaza, Encarna; Bautista-Ortín, Ana B; Ruiz-García, Yolanda; Fernández-Fernández, José I; Gil-Muñoz, Rocío

    2017-02-01

    The effect of the application of benzothiadiazole (BTH) and methyl jasmonate (MeJ) at veraison on the phenolic composition of grapes from three varieties (Monastrell, Syrah and Merlot) was studied during the ripening period, using HPLC techniques to measure flavonols, anthocyanins and tannins. The effects of the treatments differed in the three varieties, and the maximum concentration of phenolic compounds was not always reached at the end of the ripening period but some days before harvest. At the end of ripening both treated Syrah grapes only differed from control grapes in the flavonol concentration, whereas MeJ-treated Merlot grapes presented higher anthocyanin and skin tannin contents than the control and BTH-treated grapes. Only the anthocyanin content was significantly higher in treated Monastrell grapes at the moment of harvest. The results indicate that the moment of elicitor treatment should be more studied since differences between treated and control grapes were, in general greater several days before harvest in all three varieties. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  1. Proteomic Analysis of Grape Berry Cell Cultures Reveals that Developmentally Regulated Ripening Related Processes Can Be Studied Using Cultured Cells

    PubMed Central

    Sharathchandra, Ramaschandra G.; Stander, Charmaine; Jacobson, Dan; Ndimba, Bongani; Vivier, Melané A.

    2011-01-01

    Background This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first. Methodology/Principal Findings In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions. Conclusions The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry

  2. Ripening of salami: assessment of colour and aspect evolution using image analysis and multivariate image analysis.

    PubMed

    Fongaro, Lorenzo; Alamprese, Cristina; Casiraghi, Ernestina

    2015-03-01

    During ripening of salami, colour changes occur due to oxidation phenomena involving myoglobin. Moreover, shrinkage due to dehydration results in aspect modifications, mainly ascribable to fat aggregation. The aim of this work was the application of image analysis (IA) and multivariate image analysis (MIA) techniques to the study of colour and aspect changes occurring in salami during ripening. IA results showed that red, green, blue, and intensity parameters decreased due to the development of a global darker colour, while Heterogeneity increased due to fat aggregation. By applying MIA, different salami slice areas corresponding to fat and three different degrees of oxidised meat were identified and quantified. It was thus possible to study the trend of these different areas as a function of ripening, making objective an evaluation usually performed by subjective visual inspection. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Parents' information needs and influential factors when making decisions about TNF-α inhibitors.

    PubMed

    Lipstein, Ellen A; Lovell, Daniel J; Denson, Lee A; Kim, Sandra C; Spencer, Charles; Britto, Maria T

    2016-09-15

    Parents struggle when making treatment decisions for children with arthritis or other chronic conditions. Understanding their decision-making process is an essential step towards improving the decision-making experience. The objective of this study was to describe parents' information needs and the influences on their decision making about treatment with TNF-α inhibitors. Survey domains were developed based on qualitative data and cognitive interviewing. We mailed the survey to parents of children with juvenile idiopathic arthritis or inflammatory bowel disease who had initiated treatment with TNF-α inhibitors in the prior 2 years. Data were analyzed using descriptive and non-parametric statistics. Survey response rate was 54.9 %. Each item had <2 % missing responses. Parents used an array of information sources when deciding about treatment with TNF-α inhibitors. Resources other than their child's specialist were most often used to increase confidence in parents' decisions or because they wanted to know more about other people's experiences being treated with TNF-α inhibitors, rather than due to a lack of understanding. All but two (cost and route of administration) of the influential decision factors were very or extremely important to the majority of participants with factors related to long-term side effects, treatment efficacy, and disease impact being most important. This study describes parents' information needs and influential factors in treatment decision making. Results suggest that future work should be aimed at helping families weigh risks and benefits, such as through decision support interventions, as well as developing opportunities to include people beyond the family and physician in the decision-making process.

  4. Vascular endothelial growth factor inhibitors: investigational therapies for the treatment of psoriasis

    PubMed Central

    Weidemann, Anja K; Crawshaw, Ania A; Byrne, Emily; Young, Helen S

    2013-01-01

    Psoriasis is a common inflammatory autoimmune condition in which environmental factors and genetic predisposition contribute to the development of disease in susceptible individuals. Angiogenesis is known to be a key pathogenic feature of psoriasis. Local and systemic elevation of vascular endothelial growth factor (VEGF)-A has been demonstrated in the skin and plasma of patients with psoriasis and is known to correlate with improvement following some traditional psoriasis treatments. A number of VEGF inhibitors are licensed for the treatment of malignancies and eye disease and isolated case reports suggest that some individuals with psoriasis may improve when exposed to these agents. The small number of cases and lack of unified reporting measures makes it difficult to draw generalizations and underline the heterogeneity of psoriasis as a disease entity. Though not yet licensed for the treatment of psoriasis in humans, experimental data supports the potential of VEGF inhibitors to influence relevant aspects of human cell biology (such as endothelial cell differentiation) and to improve animal models of skin disease. Given the multi-factorial nature of psoriasis it is unlikely that VEGF inhibitors will be effective in all patients, however they have the potential to be a valuable addition to the therapeutic arsenal in selected cases. Current VEGF inhibitors in clinical use are associated with a number of potentially serious side effects including hypertension, left ventricular dysfunction, and gastrointestinal perforation. Such risks require careful consideration in psoriasis populations particularly in light of growing concerns linking psoriasis to increased cardiovascular risk. PMID:24101875

  5. Vascular endothelial growth factor inhibitors: investigational therapies for the treatment of psoriasis.

    PubMed

    Weidemann, Anja K; Crawshaw, Ania A; Byrne, Emily; Young, Helen S

    2013-09-26

    Psoriasis is a common inflammatory autoimmune condition in which environmental factors and genetic predisposition contribute to the development of disease in susceptible individuals. Angiogenesis is known to be a key pathogenic feature of psoriasis. Local and systemic elevation of vascular endothelial growth factor (VEGF)-A has been demonstrated in the skin and plasma of patients with psoriasis and is known to correlate with improvement following some traditional psoriasis treatments. A number of VEGF inhibitors are licensed for the treatment of malignancies and eye disease and isolated case reports suggest that some individuals with psoriasis may improve when exposed to these agents. The small number of cases and lack of unified reporting measures makes it difficult to draw generalizations and underline the heterogeneity of psoriasis as a disease entity. Though not yet licensed for the treatment of psoriasis in humans, experimental data supports the potential of VEGF inhibitors to influence relevant aspects of human cell biology (such as endothelial cell differentiation) and to improve animal models of skin disease. Given the multi-factorial nature of psoriasis it is unlikely that VEGF inhibitors will be effective in all patients, however they have the potential to be a valuable addition to the therapeutic arsenal in selected cases. Current VEGF inhibitors in clinical use are associated with a number of potentially serious side effects including hypertension, left ventricular dysfunction, and gastrointestinal perforation. Such risks require careful consideration in psoriasis populations particularly in light of growing concerns linking psoriasis to increased cardiovascular risk.

  6. Selenium delays tomato fruit ripening by inhibiting ethylene biosynthesis and enhancing the antioxidant defense system.

    PubMed

    Zhu, Zhu; Chen, Yanli; Shi, Guoqing; Zhang, Xueji

    2017-03-15

    The antioxidant activity of selenium (Se) detoxifies reactive oxygen species (ROS) in plants and animals. In the present study, we elucidated the mechanism underlying Se induced fruit development and ripening. Our study showed that foliar pretreatment with 1mgL -1 sodium selenate effectively delayed fruit ripening and maintained fruit quality. Gene expression studies revealed that the repression of ethylene biosynthetic genes 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase decreased ethylene production and respiration rate. Moreover, Se treatment probably boosted the antioxidant defense system to reduce ROS generation and membrane damage. The enhanced antioxidative effect was attributed to higher glutathione content and increased activity of enzymes such as glutathione peroxidase and glutathione reductase. The upregulation of respiratory burst oxidase homologue genes in tomato fruit may also contribute to the enhanced antioxidative effect. Selenium treatment represents a promising strategy for delaying ripening and extending the shelf life of tomato fruit. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Expression patterns of cell wall-modifying genes from banana during fruit ripening and in relationship with finger drop

    PubMed Central

    Mbéguié-A-Mbéguié, D.; Hubert, O.; Baurens, F. C.; Matsumoto, T.; Chillet, M.; Fils-Lycaon, B.; Sidibé-Bocs, S.

    2009-01-01

    Few molecular studies have been devoted to the finger drop process that occurs during banana fruit ripening. Recent studies revealed the involvement of changes in the properties of cell wall polysaccharides in the pedicel rupture area. In this study, the expression of cell-wall modifying genes was monitored in peel tissue during post-harvest ripening of Cavendish banana fruit, at median area (control zone) and compared with that in the pedicel rupture area (drop zone). To this end, three pectin methylesterase (PME) and seven xyloglucan endotransglycosylase/hydrolase (XTH) genes were isolated. The accumulation of their mRNAs and those of polygalaturonase, expansin, and pectate lyase genes already isolated from banana were examined. During post-harvest ripening, transcripts of all genes were detected in both zones, but accumulated differentially. MaPME1, MaPG1, and MaXTH4 mRNA levels did not change in either zone. Levels of MaPME3 and MaPG3 mRNAs increased greatly only in the control zone and at the late ripening stages. For other genes, the main molecular changes occurred 1–4 d after ripening induction. MaPME2, MaPEL1, MaPEL2, MaPG4, MaXTH6, MaXTH8, MaXTH9, MaEXP1, MaEXP4, and MaEXP5 accumulated highly in the drop zone, contrary to MaXTH3 and MaXTH5, and MaEXP2 throughout ripening. For MaPG2, MaXET1, and MaXET2 genes, high accumulation in the drop zone was transient. The transcriptional data obtained from all genes examined suggested that finger drop and peel softening involved similar mechanisms. These findings also led to the proposal of a sequence of molecular events leading to finger drop and to suggest some candidates. PMID:19357434

  8. Jasmonate signalling pathway in strawberry: Genome-wide identification, molecular characterization and expression of JAZs and MYCs during fruit development and ripening.

    PubMed

    Garrido-Bigotes, Adrián; Figueroa, Nicolás E; Figueroa, Pablo M; Figueroa, Carlos R

    2018-01-01

    Jasmonates (JAs) are signalling molecules involved in stress responses, development and secondary metabolism biosynthesis, although their roles in fleshy-fruit development and ripening processes are not well known. In strawberry fruit, it has been proposed that JAs could regulate the early development through the activation of the JAs biosynthesis. Moreover, it has been reported that JA treatment increases anthocyanin content in strawberry fruit involving the bioactive jasmonate biosynthesis. Nevertheless, JA signalling pathway, of which main components are the COI1-JAZ co-receptor and the MYC transcription factors (TFs), has not been characterized in strawberry until now. Here we identified and characterized the woodland strawberry (Fragaria vesca) JAZ and MYC genes as well as studied their expression during development and ripening stages in commercial strawberry (Fragaria × ananassa) fruit. We described twelve putative JAZ proteins and two MYC TFs, which showed high conservation with respect to their orthologs in Arabidopsis thaliana and in other fleshy-fruit species such as Malus × domestica, Vitis vinifera and Solanum lycopersicum as revealed by gene synteny and phylogenetic analyses. Noteworthy, their expression levels exhibited a significant decrease from fruit development to ripening stages in F. × ananassa, along with others of the JA signalling-related genes such as FaNINJA and FaJAMs, encoding for negative regulators of JA responses. Moreover, we found that main JA signalling-related genes such as FaMYC2, and FaJAZ1 are promptly induced by JA treatment at early times in F. × ananassa fruit. These results suggest the conservation of the canonical JA signalling pathway in strawberry and a possible role of this pathway in early strawberry fruit development, which also correlates negatively with the beginning of the ripening process.

  9. Jasmonate signalling pathway in strawberry: Genome-wide identification, molecular characterization and expression of JAZs and MYCs during fruit development and ripening

    PubMed Central

    Figueroa, Nicolás E.; Figueroa, Pablo M.

    2018-01-01

    Jasmonates (JAs) are signalling molecules involved in stress responses, development and secondary metabolism biosynthesis, although their roles in fleshy-fruit development and ripening processes are not well known. In strawberry fruit, it has been proposed that JAs could regulate the early development through the activation of the JAs biosynthesis. Moreover, it has been reported that JA treatment increases anthocyanin content in strawberry fruit involving the bioactive jasmonate biosynthesis. Nevertheless, JA signalling pathway, of which main components are the COI1-JAZ co-receptor and the MYC transcription factors (TFs), has not been characterized in strawberry until now. Here we identified and characterized the woodland strawberry (Fragaria vesca) JAZ and MYC genes as well as studied their expression during development and ripening stages in commercial strawberry (Fragaria × ananassa) fruit. We described twelve putative JAZ proteins and two MYC TFs, which showed high conservation with respect to their orthologs in Arabidopsis thaliana and in other fleshy-fruit species such as Malus × domestica, Vitis vinifera and Solanum lycopersicum as revealed by gene synteny and phylogenetic analyses. Noteworthy, their expression levels exhibited a significant decrease from fruit development to ripening stages in F. × ananassa, along with others of the JA signalling-related genes such as FaNINJA and FaJAMs, encoding for negative regulators of JA responses. Moreover, we found that main JA signalling-related genes such as FaMYC2, and FaJAZ1 are promptly induced by JA treatment at early times in F. × ananassa fruit. These results suggest the conservation of the canonical JA signalling pathway in strawberry and a possible role of this pathway in early strawberry fruit development, which also correlates negatively with the beginning of the ripening process. PMID:29746533

  10. Different Citrus rootstocks present high dissimilarities in their antioxidant activity and vitamins content according to the ripening stage.

    PubMed

    Cardeñosa, Vanessa; Barros, Lillian; Barreira, João C M; Arenas, Francisco; Moreno-Rojas, José M; Ferreira, Isabel C F R

    2015-02-01

    "Lane Late" sweet orange grafted on six different citrus rootstocks and grown in the Guadalquivir valley (Seville, Spain) were picked at different ripening stages in two consecutive seasons to characterize their antioxidant activity (free radicals scavenging activity, reducing power and lipid peroxidation inhibition) and quantify their main antioxidant compounds (vitamin E and vitamin C). Linear discriminant analysis and 2-way ANOVA were applied to compare the effects induced by citrus rootstock and ripening stage. The results showed that differences in antioxidant activity and related compounds are mainly dependent on the citrus rootstock, despite ripening stage had also some particular effects. Changes observed in 2012 showed less marked differences among the citrus rootstock. Nevertheless, Cleopatra rootstock showed the highest antioxidant activity in both years, indicating that an increase in its cultivation might be a good solution to sweet orange farmers. Concerning the ripening stage, samples collected in January presented higher vitamin contents, while those collected in April showed higher antioxidant activity. This result allows deciding the harvesting period according to the desired effect. Copyright © 2014 Elsevier GmbH. All rights reserved.

  11. Virus-Induced Gene Silencing of the Eggplant Chalcone Synthase Gene during Fruit Ripening Modifies Epidermal Cells and Gravitropism.

    PubMed

    Wang, Cuicui; Fu, Daqi

    2018-03-21

    Eggplant ( Solanum melongena L.) fruits accumulate flavonoids in their cuticle and epidermal cells during ripening. Although many mutants available in model plant species, such as Arabidopsis thaliana and Medicago truncatula, are enabling the intricacies of flavonoid-related physiology to be deduced, the mechanisms whereby flavonoids influence eggplant fruit physiology are unknown. Virus-induced gene silencing (VIGS) is a reliable tool for the study of flavonoid function in fruit, and in this study, we successfully applied this technique to downregulate S. melongena chalcone synthase gene ( SmCHS) expression during eggplant fruit ripening. In addition to the expected change in fruit color attributable to a lack of anthocyanins, several other modifications, including differences in epidermal cell size and shape, were observed in the different sectors. We also found that silencing of CHS gene expression was associated with a negative gravitropic response in eggplant fruits. These observations indicate that epidermal cell expansion during ripening is dependent upon CHS expression and that there may be a relationship between CHS expression and gravitropism during eggplant fruit ripening.

  12. Survival of Escherichia coli O157:H7 during the manufacture and ripening of Cacioricotta goat cheese.

    PubMed

    Ioanna, F; Quaglia, N C; Storelli, M M; Castiglia, D; Goffredo, E; Storelli, A; De Rosa, M; Normanno, G; Jambrenghi, A Caputi; Dambrosio, A

    2018-04-01

    The aim of this study was to assess the growth and survival of Escherichia coli O157:H7 during the manufacturing and ripening of Cacioricotta goat cheese. Goat milk was artificially contaminated with E. coli O157:H7 and the bacterial load was monitored from production up to 90 days of ripening. Goat milk was inoculated with 10 2  cfu ml -1 of E. coli O157:H7 and the bacterial count of the curd at time zero was 2.31 log 10  cfu g -1 . During the first day of ripening, the bacterial load has increased to 5.73 log 10  cfu g -1 to more than 6.20 log 10  cfu g -1 during the first week. The bacterial load remained constant up to 28 days and then slightly decreased until the end of ripening, with values of a w and pH of 0.88 and 5.41 respectively. The results of this study highlighted that E. coli O157:H7 is able to survive the manufacturing process and they suggest that the 90-day period of ripening alone is insufficient to remove E. coli O157:H7 in contaminated Cacioricotta goat cheese. Moreover, these results support the assumption that the presence of a low contamination of milk with E. coli O157:H7 could represent a potential source of infection and a threat to consumers. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Effect of microgravity simulation using 3D clinostat on cavendish banana (Musa acuminata AAA Group) ripening process

    NASA Astrophysics Data System (ADS)

    Dwivany, Fenny Martha; Esyanti, Rizkita R.; Prapaisie, Adeline; Puspa Kirana, Listya; Latief, Chunaeni; Ginaldi, Ari

    2016-11-01

    The objective of the research was to determine the effect of microgravity simulation by 3D clinostat on Cavendish banana (Musa acuminata AAA group) ripening process. In this study, physical, physiological changes as well as genes expression were analysed. The result showed that in microgravity simulation condition ripening process in banana was delayed and the MaACOl, MaACSl and MaACS5 gene expression were affected.

  14. Proteasome inhibitors enhance endothelial thrombomodulin expression via induction of Krüppel-like transcription factors

    PubMed Central

    Hiroi, Toyoko; Deming, Clayton B.; Zhao, Haige; Hansen, Baranda S.; Arkenbout, Elisabeth K.; Myers, Thomas J.; McDevitt, Michael A.; Rade, Jeffrey J.

    2009-01-01

    Objective Impairment of the thrombomodulin-protein C anticoagulant pathway has been implicated in pathologic thrombosis associated with malignancy. Patients who receive proteasome inhibitors as part of their chemotherapeutic regimen appear to be at decreased risk for thromboembolic events. We investigated the effects of proteasome inhibitors on endothelial thrombomodulin expression and function. Methods and Results Proteasome inhibitors as a class markedly induced the expression thrombomodulin and enhanced the protein C activating capacity of endothelial cells. Thrombomodulin upregulation was independent of NF-κB signaling, a principal target of proteasome inhibitors, but was instead a direct consequence of increased expression of the Krüppel-like transcription factors, KLF2 and KLF4. These effects were confirmed in vivo, where systemic administration of a proteasome inhibitor enhanced thrombomodulin expression that was paralleled by changes in the expression of KLF2 and KLF4. Conclusions These findings identify a novel mechanism of action of proteasome inhibitors that may help to explain their clinically observed thromboprotective effects. PMID:19661484

  15. Linking hormonal profiles with variations in sugar and anthocyanin contents during the natural development and ripening of sweet cherries.

    PubMed

    Teribia, Natalia; Tijero, Verónica; Munné-Bosch, Sergi

    2016-12-25

    Sweet cherries are highly appreciated by consumers worldwide and are usually cold-stored during postharvest to prevent over-ripening before distribution to the market. Sweet cherry is a non-climacteric fruit, for which ripening is known to be regulated by abscisic acid. Here we aimed to examine the hormone profiles, including measurements of abscisic acid, auxins, cytokinins and gibberellins by ultrahigh performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS), in relation to variations in sugar and anthocyanin contents, during growth and ripening of this fruit. Hormonal profiling revealed that indole-3-acetic acid, GA 1 and trans-zeatin levels decreased at early stages of fruit development, while GA 3 levels decreased at early stages but also later, once anthocyanin accumulation started. Conversely, abscisic acid levels rose significantly once the fruit started to synthetize anthocyanins, and isopentenyladenosine levels also increased during the ripening of sweet cherries. A strong negative correlation was found between GA 4 levels and both fruit biomass and anthocyanin levels, and between the levels of trans-zeatin and both fruit biomass and total sugar contents. In contrast, abscisic acid and isopentenyladenosine levels correlated positively with fruit biomass, anthocyanin and total soluble sugar content. Results suggest that auxins, cytokinins and gibberellins may act coordinately with abscisic acid in the regulation of sweet cherry development and ripening. Furthermore, it is shown that hormonal profile measurements by UHPLC-MS/MS may be a helpful tool to elucidate the timing of action of each specific hormonal compound during ripening, which has important applications in the agri-food biotechnological sector. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Pectic-β(1,4)-galactan, extensin and arabinogalactan–protein epitopes differentiate ripening stages in wine and table grape cell walls

    PubMed Central

    Moore, John P.; Fangel, Jonatan U.; Willats, William G. T.; Vivier, Melané A.

    2014-01-01

    Background and Aims Cell wall changes in ripening grapes (Vitis vinifera) have been shown to involve re-modelling of pectin, xyloglucan and cellulose networks. Newer experimental techniques, such as molecular probes specific for cell wall epitopes, have yet to be extensively used in grape studies. Limited general information is available on the cell wall properties that contribute to texture differences between wine and table grapes. This study evaluates whether profiling tools can detect cell wall changes in ripening grapes from commercial vineyards. Methods Standard sugar analysis and infra-red spectroscopy were used to examine the ripening stages (green, véraison and ripe) in grapes collected from Cabernet Sauvignon and Crimson Seedless vineyards. Comprehensive microarray polymer profiling (CoMPP) analysis was performed on cyclohexanediaminetetraacetic acid (CDTA) and NaOH extracts of alcohol-insoluble residue sourced from each stage using sets of cell wall probes (mAbs and CBMs), and the datasets were analysed using multivariate software. Key Results The datasets obtained confirmed previous studies on cell wall changes known to occur during grape ripening. Probes for homogalacturonan (e.g. LM19) were enriched in the CDTA fractions of Crimson Seedless relative to Cabernet Sauvignon grapes. Probes for pectic-β-(1,4)-galactan (mAb LM5), extensin (mAb LM1) and arabinogalactan proteins (AGPs, mAb LM2) were strongly correlated with ripening. From green stage to véraison, a progressive reduction in pectic-β-(1,4)-galactan epitopes, present in both pectin-rich (CDTA) and hemicellulose-rich (NaOH) polymers, was observed. Ripening changes in AGP and extensin epitope abundance also were found during and after véraison. Conclusions Combinations of cell wall probes are able to define distinct ripening phases in grapes. Pectic-β-(1,4)-galactan epitopes decreased in abundance from green stage to véraison berries. From véraison there was an increase in abundance of

  17. Tissue specific localization of pectin-Ca²⁺ cross-linkages and pectin methyl-esterification during fruit ripening in tomato (Solanum lycopersicum).

    PubMed

    Hyodo, Hiromi; Terao, Azusa; Furukawa, Jun; Sakamoto, Naoya; Yurimoto, Hisayoshi; Satoh, Shinobu; Iwai, Hiroaki

    2013-01-01

    Fruit ripening is one of the developmental processes accompanying seed development. The tomato is a well-known model for studying fruit ripening and development, and the disassembly of primary cell walls and the middle lamella, such as through pectin de-methylesterified by pectin methylesterase (PE) and depolymerization by polygalacturonase (PG), is generally accepted to be one of the major changes that occur during ripening. Although many reports of the changes in pectin during tomato fruit ripening are focused on the relation to softening of the pericarp or the Blossom-end rot by calcium (Ca²⁺) deficiency disorder, the changes in pectin structure and localization in each tissues during tomato fruit ripening is not well known. In this study, to elucidate the tissue-specific role of pectin during fruit development and ripening, we examined gene expression, the enzymatic activities involved in pectin synthesis and depolymerisation in fruit using biochemical and immunohistochemical analyses, and uronic acids and calcium (Ca)-bound pectin were determined by secondary ion-microprobe mass spectrometry. These results show that changes in pectin properties during fruit development and ripening have tissue-specific patterns. In particular, differential control of pectin methyl-esterification occurs in each tissue. Variations in the cell walls of the pericarp are quite different from that of locular tissues. The Ca-binding pectin and hairy pectin in skin cell layers are important for intercellular and tissue-tissue adhesion. Maintenance of the globular form and softening of tomato fruit may be regulated by the arrangement of pectin structures in each tissue.

  18. Non-genetic risk factors in haemophilia A inhibitor management - the danger theory and the use of animal models.

    PubMed

    Lövgren, K M; Søndergaard, H; Skov, S; Wiinberg, B

    2016-09-01

    In haemophilia A (HA) management, antidrug antibodies, or inhibitors, are a serious complication that renders factor VIII (FVIII) replacement therapy ineffective, increases morbidity and reduces quality of life for affected patients. Inhibitor development aetiology is multifactorial and covers both genetic and therapy related risk factors. Many therapy-related risk factors have proven difficult to confirm due to several confounding factors and the small study populations available. However, clinical studies indicate that e.g. on-demand treatment and surgery affect inhibitor development, and explanations for this association are being investigated. A potential explanation is the danger signal effect, where the immune response is activated by endogenous or exogenous danger or damage signals present at the time and site of FVIII administration. The danger theory explains how alarm signals from stressed, injured or dying cells can activate an immune reaction, without the involvement of foreign antigens. Bleeds, trauma, surgery or concomitant infection could be events initiating danger signalling in HA patients, resulting in an immune reaction towards administered FVIII that otherwise would pass unnoticed. This role of danger in HA inhibitor formation has previously been suggested, but a thorough discussion of this subject is lacking. The present review will discuss the potential role of danger signals in haemophilia and inhibitor development, with focus on treatment related risk factors with a suspected danger signal aetiology; on-demand treatment, treatment during major bleeds or surgery, and treatment during infection or vaccination. Clinical studies as well as animal experiments addressing these factors will be reviewed. © 2016 John Wiley & Sons Ltd.

  19. Preoperative ripening of the cervix before operative hysteroscopy.

    PubMed

    Al-Fozan, Haya; Firwana, Belal; Al Kadri, Hanan; Hassan, Samar; Tulandi, Togas

    2015-04-23

    Hysteroscopy is an operation in which the gynaecologist examines the uterine cavity using a small telescopic instrument (hysteroscope) inserted via the vagina and the cervix. Almost 50% of hysteroscopic complications are related to difficulty with cervical entry. Potential complications include cervical tears, creation of a false passage, perforation, bleeding, or simply difficulty in entering the internal os (between the cervix and the uterus) with the hysteroscope. These complications may possibly be reduced with adequate preparation of the cervix (cervical ripening) prior to hysteroscopy. Cervical ripening agents include oral or vaginal prostaglandin, which can be synthetic (e.g misoprostol) or natural (e.g. dinoprostone) and vaginal osmotic dilators, which can be naturally occurring (e.g. laminaria) or synthetic. To determine whether preoperative cervical preparation facilitates cervical dilatation and reduces the complications of operative hysteroscopy in women undergoing the procedure for any condition. In August 2014 we searched sources including the Menstrual Disorders and Subfertility Group (MDSG) Trials Register, Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, PsycINFO, CINAHL, ClinicalTrials.gov and reference lists of relevant articles. We searched for published and unpublished studies in any language. Two review authors independently selected randomised controlled trials (RCTs) of cervical ripening agents used before operative hysteroscopy in pre- and postmenopausal women. Cervical ripening agents could be compared to each other, placebo or no treatment. Data extraction and quality assessment were conducted independently by two review authors. The primary review outcomes were effectiveness of cervical dilatation (defined as the proportion of women requiring mechanical cervical dilatation) and intraoperative complications. Secondary outcomes were mean time required to dilate the cervix, preoperative pain, cervical width

  20. The Citrus ABA signalosome: identification and transcriptional regulation during sweet orange fruit ripening and leaf dehydration.

    PubMed

    Romero, Paco; Lafuente, María T; Rodrigo, María J

    2012-08-01

    The abscisic acid (ABA) signalling core in plants include the cytosolic ABA receptors (PYR/PYL/RCARs), the clade-A type 2C protein phosphatases (PP2CAs), and the subclass III SNF1-related protein kinases 2 (SnRK2s). The aim of this work was to identify these ABA perception system components in sweet orange and to determine the influence of endogenous ABA on their transcriptional regulation during fruit development and ripening, taking advantage of the comparative analysis between a wild-type and a fruit-specific ABA-deficient mutant. Transcriptional changes in the ABA signalosome during leaf dehydration were also studied. Six PYR/PYL/RCAR, five PP2CA, and two subclass III SnRK2 genes, homologous to those of Arabidopsis, were identified in the Citrus genome. The high degree of homology and conserved motifs for protein folding and for functional activity suggested that these Citrus proteins are bona fide core elements of ABA perception in orange. Opposite expression patterns of CsPYL4 and CsPYL5 and ABA accumulation were found during ripening, although there were few differences between varieties. In contrast, changes in expression of CsPP2CA genes during ripening paralleled those of ABA content and agreeed with the relevant differences between wild-type and mutant fruit transcript accumulation. CsSnRK2 gene expression continuously decreased with ripening and no remarkable differences were found between cultivars. Overall, dehydration had a minor effect on CsPYR/PYL/RCAR and CsSnRK2 expression in vegetative tissue, whereas CsABI1, CsAHG1, and CsAHG3 were highly induced by water stress. The global results suggest that responsiveness to ABA changes during citrus fruit ripening, and leaf dehydration was higher in the CsPP2CA gene negative regulators than in the other ABA signalosome components.

  1. The Citrus ABA signalosome: identification and transcriptional regulation during sweet orange fruit ripening and leaf dehydration

    PubMed Central

    Rodrigo, María J.

    2012-01-01

    The abscisic acid (ABA) signalling core in plants include the cytosolic ABA receptors (PYR/PYL/RCARs), the clade-A type 2C protein phosphatases (PP2CAs), and the subclass III SNF1-related protein kinases 2 (SnRK2s). The aim of this work was to identify these ABA perception system components in sweet orange and to determine the influence of endogenous ABA on their transcriptional regulation during fruit development and ripening, taking advantage of the comparative analysis between a wild-type and a fruit-specific ABA-deficient mutant. Transcriptional changes in the ABA signalosome during leaf dehydration were also studied. Six PYR/PYL/RCAR, five PP2CA, and two subclass III SnRK2 genes, homologous to those of Arabidopsis, were identified in the Citrus genome. The high degree of homology and conserved motifs for protein folding and for functional activity suggested that these Citrus proteins are bona fide core elements of ABA perception in orange. Opposite expression patterns of CsPYL4 and CsPYL5 and ABA accumulation were found during ripening, although there were few differences between varieties. In contrast, changes in expression of CsPP2CA genes during ripening paralleled those of ABA content and agreeed with the relevant differences between wild-type and mutant fruit transcript accumulation. CsSnRK2 gene expression continuously decreased with ripening and no remarkable differences were found between cultivars. Overall, dehydration had a minor effect on CsPYR/PYL/RCAR and CsSnRK2 expression in vegetative tissue, whereas CsABI1, CsAHG1, and CsAHG3 were highly induced by water stress. The global results suggest that responsiveness to ABA changes during citrus fruit ripening, and leaf dehydration was higher in the CsPP2CA gene negative regulators than in the other ABA signalosome components. PMID:22888124

  2. Evidences for Chlorogenic Acid — A Major Endogenous Polyphenol Involved in Regulation of Ripening and Senescence of Apple Fruit

    PubMed Central

    Xi, Yu; Cheng, Dai; Zeng, Xiangquan; Cao, Jiankang; Jiang, Weibo

    2016-01-01

    To learn how the endogenous polyphenols may play a role in fruit ripening and senescence, apple pulp discs were used as a model to study the influences of chlorogenic acid (CHA, a major polyphenol in apple pulp) on fruit ripening and senescence. Apple (‘Golden Delicious’) pulp discs prepared from pre-climacteric fruit were treated with 50 mg L-1 CHA and incubated in flasks with 10 mM MES buffer (pH 6.0, 11% sorbitol). Compared to the control samples, treatment with CHA significantly reduced ethylene production and respiration rate, and enhanced levels of firmness and soluble solids content of the pulp discs during incubation at 25°C. These results suggested that CHA could retard senescence of the apple pulp discs. Proteomics analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry (MALDI-TOF/TOF) revealed that the expressions of several key proteins correlated to fruit ripening and senescence were affected by the treatment with CHA. Further study showed that treating the pulp discs with CHA remarkably reduced levels of lipoxygenase, β-galactosidase, NADP-malic enzyme, and enzymatic activities of lipoxygenase and UDP-glucose pyrophosphorylase, all of which are known as promoters of fruit ripening and senescence. These results could provide new insights into the functions of endogenous phenolic compounds in fruit ripening and senescence. PMID:26756813

  3. Sarcoidosis as an adverse effect of tumor necrosis factor inhibitors.

    PubMed

    Cathcart, Shelley; Sami, Naveed; Elewski, Boni

    2012-05-01

    Tumor necrosis factor inhibitors are valuable tools for dermatologists. As their use increases, rare adverse events are more likely to be encountered. We describe one patient who developed sarcoidosis while being treated for psoriasis with etanercept. We sought to review to previously reported cases and further characterize the nature of this reaction. A literature search was performed with the key words "sarcoidosis, sarcoid, etanercept, infliximab, adalimumab, granulomatous, and drug reaction." All relevant cases in the English language were included and evaluated for demographic data, duration of therapy prior to developing sarcoid, duration of sarcoid signs/symptoms, treatments used and time to resolution after discontinuation of the drug. Including the present case, there are 34 cases of sarcoidosis developing during anti-tumor necrosis factor therapy. All previously reported cases were patients with a primarily rheumatologic diagnosis. In all but one case, discontinuation of the drug resulted in complete resolution of symptoms. The lung and surrounding lymph nodes were the areas most commonly affected. The average amount of time between initiation of therapy and onset of symptoms was 22 months. The average time to resolution of symptoms after discontinuation of the drug was 5.2 months. This is a retrospective case review. These data indicated that sarcoid is a possible adverse effect of tumor necrosis factor inhibitor therapy that should be noted by dermatologists using these drugs. While it has been reported in the rheumatology literature, it may be under-recognized by dermatologists.

  4. Safety of 4-factor prothrombin complex concentrate (4F-PCC) for emergent reversal of factor Xa inhibitors.

    PubMed

    Tao, Jing; Bukanova, Elena N; Akhtar, Shamsuddin

    2018-01-01

    Although factor Xa inhibitors have become a popular choice for chronic oral anticoagulation, effective drug reversal remains difficult due to a lack of specific antidote. Currently, 4-factor prothrombin complex concentrate (4F-PCC) is considered the treatment of choice for factor Xa inhibitor-related major bleeding. However, safety of 4F-PCC and its risk of thrombosis when used for this off-label purpose remain unclear. The purpose of this retrospective study is to determine the rate of thromboembolism when 4F-PCC is used for the emergent reversal of factor Xa inhibitors. We conducted a single-center retrospective review of medical records between 2013 and 2017. Patients were included if they received 4F-PCC to reverse rivaroxaban, apixaban, or edoxaban for emergent invasive procedures or during episodes of major bleeding defined as bleeding with hemodynamic instability, fall in hemoglobin of 2 g/dL, or bleeding requiring blood transfusion. Thrombotic events including myocardial infarction, pulmonary embolism, deep vein thrombosis, cerebral vascular accident, and arterial thrombosis of the limb or mesentery were recorded if they occurred within 14 days of 4F-PCC administration. Data was analyzed using point and interval estimation to approximate the rate and confidence interval of thromboembolic events. Forty-three patients were identified in our review. Doses of 4F-PCC were determined by the treating physician and mainly ranged from 25 to 50 IU/kg. Twenty-two patients (51.2%) received both sequential compression devices (SCDs) and subcutaneous heparin for DVT prophylaxis. Twenty-one patients (48.8%) were placed on SCDs only. Three patients received concomitant FFP. Thrombotic events within 14 days of 4F-PCC administration occurred in 1 out of 43 patients (2.1%, 95% CI [0.1-12.3]). This thrombotic event was an upper extremity DVT which occurred 1 day after the patient received 1325 IU (25 IU/kg) of 4F-PCC to reverse rivaroxaban for traumatic intracranial

  5. Identification of Differentially Expressed Genes Associated with Apple Fruit Ripening and Softening by Suppression Subtractive Hybridization

    PubMed Central

    Zhang, Zongying; Jiang, Shenghui; Wang, Nan; Li, Min; Ji, Xiaohao; Sun, Shasha; Liu, Jingxuan; Wang, Deyun; Xu, Haifeng; Qi, Sumin; Wu, Shujing; Fei, Zhangjun; Feng, Shouqian; Chen, Xuesen

    2015-01-01

    Apple is one of the most economically important horticultural fruit crops worldwide. It is critical to gain insights into fruit ripening and softening to improve apple fruit quality and extend shelf life. In this study, forward and reverse suppression subtractive hybridization libraries were generated from ‘Taishanzaoxia’ apple fruits sampled around the ethylene climacteric to isolate ripening- and softening-related genes. A set of 648 unigenes were derived from sequence alignment and cluster assembly of 918 expressed sequence tags. According to gene ontology functional classification, 390 out of 443 unigenes (88%) were assigned to the biological process category, 356 unigenes (80%) were classified in the molecular function category, and 381 unigenes (86%) were allocated to the cellular component category. A total of 26 unigenes differentially expressed during fruit development period were analyzed by quantitative RT-PCR. These genes were involved in cell wall modification, anthocyanin biosynthesis, aroma production, stress response, metabolism, transcription, or were non-annotated. Some genes associated with cell wall modification, anthocyanin biosynthesis and aroma production were up-regulated and significantly correlated with ethylene production, suggesting that fruit texture, coloration and aroma may be regulated by ethylene in ‘Taishanzaoxia’. Some of the identified unigenes associated with fruit ripening and softening have not been characterized in public databases. The results contribute to an improved characterization of changes in gene expression during apple fruit ripening and softening. PMID:26719904

  6. Nontraumatic spinal subdural hematoma complicating direct factor Xa inhibitor treatment (rivaroxaban): a challenging management.

    PubMed

    Dargazanli, Cyril; Lonjon, Nicolas; Gras-Combe, Guillaume

    2016-05-01

    We report on a 72-year-old male patient who developed a nontraumatic spinal subdural hematoma (SSDH) during rivaroxaban therapy, a relatively new orally administered direct factor Xa inhibitor. The patient sustained a sudden onset of interscapular pain, followed by gait impairment and paraplegia. Magnetic resonance imaging (MRI) of the spine demonstrated SSDH from T6 to T8. Laboratory tests revealed a high rivaroxaban level, associated with a major hemorrhagic risk. Surgery was, therefore, performed the following morning, after normalization of coagulation parameters. Determining the time of safe surgery remains challenging when hemorrhagic complications happen with direct factor Xa inhibitor, especially when neurological prognosis is engaged. Spinal subdural hematoma has not previously been reported following rivaroxaban therapy.

  7. The role of FaBG3 in fruit ripening and B. cinerea fungal infection of strawberry.

    PubMed

    Li, Qian; Ji, Kai; Sun, Yufei; Luo, Hao; Wang, Hongqing; Leng, Ping

    2013-10-01

    In plants, β-glucosidases (BG) have been implicated in developmental and pathogen defense, and are thought to take part in abscisic acid (ABA) synthesis via hydrolysis of ABA glucose ester to release active ABA; however, there is no genetic evidence for the role of BG genes in ripening and biotic/abiotic stress in fruits. To clarify the role of BG genes in fruit, eight Fa/FvBG genes encoding β-glucosidase were isolated using information from the GenBank strawberry nucleotide database. Of the Fa/FvBG genes examined, expression of FaBG3 was the highest, showing peaks at the mature stage, coincident with the changes observed in ABA content. To verify the role of this gene, we suppressed the expression of FaBG3 via inoculation with Agrobacterium tumefaciens containing tobacco rattle virus carrying a FaBG3 fragment (RNAi). The expression of FaBG3 in FaBG3-RNAi-treated fruit was markedly reduced, and the ABA content was lower than that of the control. FaBG3-RNAi-treated fruit did not exhibit full ripening, and were firmer, had lower sugar content, and were pale compared with the control due to down-regulation of ripening-related genes. FaBG3-RNAi-treated fruit with reduced ABA levels were much more resistant to Botrytis cinerea fungus but were more sensitive to dehydration stress than control fruit. These results indicate that FaBG3 may play key roles in fruit ripening, dehydration stress and B. cinerea fungal infection in strawberries via modulation of ABA homeostasis and transcriptional regulation of ripening-related genes. © 2013 The Authors The Plant Journal © 2013 John Wiley & Sons Ltd.

  8. Quantitative proteomic analysis of bacterial enzymes released in cheese during ripening.

    PubMed

    Jardin, Julien; Mollé, Daniel; Piot, Michel; Lortal, Sylvie; Gagnaire, Valérie

    2012-04-02

    Due to increasingly available bacterial genomes in databases, proteomic tools have recently been used to screen proteins expressed by micro-organisms in food in order to better understand their metabolism in situ. While the main objective is the systematic identification of proteins, the next step will be to bridge the gap between identification and quantification of these proteins. For that purpose, a new mass spectrometry-based approach was applied, using isobaric tagging reagent for quantitative proteomic analysis (iTRAQ), which are amine specific and yield labelled peptides identical in mass. Experimental Swiss-type cheeses were manufactured from microfiltered milk using Streptococcus thermophilus ITG ST20 and Lactobacillus helveticus ITG LH1 as lactic acid starters. At three ripening times (7, 20 and 69 days), cheese aqueous phases were extracted and enriched in bacterial proteins by fractionation. Each sample, standardised in protein amount prior to proteomic analyses, was: i) analysed by 2D-electrophoresis for qualitative analysis and ii) submitted to trypsinolysis, and labelled with specific iTRAQ tag, one per ripening time. The three labelled samples were mixed together and analysed by nano-LC coupled on-line with ESI-QTOF mass spectrometer. Thirty proteins, both from bacterial or bovine origin, were identified and efficiently quantified. The free bacterial proteins detected were enzymes from the central carbon metabolism as well as stress proteins. Depending on the protein considered, the quantity of these proteins in the cheese aqueous extract increased from 2.5 to 20 fold in concentration from day 7 to day 69 of ripening. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Factor Xa inhibitors versus vitamin K antagonists for preventing cerebral or systemic embolism in patients with atrial fibrillation.

    PubMed

    Bruins Slot, Karsten M H; Berge, Eivind

    2013-08-08

    Anticoagulant treatment with vitamin K antagonists (VKAs) is aimed at preventing thromboembolic complications and has been the therapy of choice for most people with non-valvular atrial fibrillation (AF) for many decades. A new class of anticoagulants, the factor Xa inhibitors, appear to have several pharmacological and practical advantages over VKAs. To assess the effectiveness and safety of treatment with factor Xa inhibitors versus VKAs for the prevention of cerebral or systemic embolic events in people with AF. We searched the trials registers of the Cochrane Stroke Group and the Cochrane Heart Group (June 2012), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2012, Issue 10), MEDLINE (1950 to April 2013) and EMBASE (1980 to April 2013). In an effort to identify further published, unpublished and ongoing trials we searched trials registers and Google Scholar (July 2012). We also screened reference lists and contacted pharmaceutical companies, authors and sponsors of relevant published trials. Randomised controlled trials that directly compared the effects of long-term treatment (more than four weeks) with factor Xa inhibitors and VKAs for the prevention of cerebral and systemic embolism in patients with AF. We included patients with and without a previous stroke or TIA. The primary efficacy outcome was the composite endpoint of all strokes and other systemic embolic events. Two authors independently assessed trial quality and the risk of bias, and extracted data. We calculated a weighted estimate of the typical treatment effect across trials using the odds ratio (OR) with 95% confidence interval (CI) by means of a fixed-effect model. However, in the case of moderate or high heterogeneity of treatment effects, we used a random-effects model to compare the overall treatment effects and performed a pre-specified sensitivity analysis excluding any fully open-label studies. We included data from 42,084 participants randomised into

  10. Small molecule inhibitors of Late SV40 Factor (LSF) abrogate hepatocellular carcinoma (HCC): Evaluation using an endogenous HCC model.

    PubMed

    Rajasekaran, Devaraja; Siddiq, Ayesha; Willoughby, Jennifer L S; Biagi, Jessica M; Christadore, Lisa M; Yunes, Sarah A; Gredler, Rachel; Jariwala, Nidhi; Robertson, Chadia L; Akiel, Maaged A; Shen, Xue-Ning; Subler, Mark A; Windle, Jolene J; Schaus, Scott E; Fisher, Paul B; Hansen, Ulla; Sarkar, Devanand

    2015-09-22

    Hepatocellular carcinoma (HCC) is a lethal malignancy with high mortality and poor prognosis. Oncogenic transcription factor Late SV40 Factor (LSF) plays an important role in promoting HCC. A small molecule inhibitor of LSF, Factor Quinolinone Inhibitor 1 (FQI1), significantly inhibited human HCC xenografts in nude mice without harming normal cells. Here we evaluated the efficacy of FQI1 and another inhibitor, FQI2, in inhibiting endogenous hepatocarcinogenesis. HCC was induced in a transgenic mouse with hepatocyte-specific overexpression of c-myc (Alb/c-myc) by injecting N-nitrosodiethylamine (DEN) followed by FQI1 or FQI2 treatment after tumor development. LSF inhibitors markedly decreased tumor burden in Alb/c-myc mice with a corresponding decrease in proliferation and angiogenesis. Interestingly, in vitro treatment of human HCC cells with LSF inhibitors resulted in mitotic arrest with an accompanying increase in CyclinB1. Inhibition of CyclinB1 induction by Cycloheximide or CDK1 activity by Roscovitine significantly prevented FQI-induced mitotic arrest. A significant induction of apoptosis was also observed upon treatment with FQI. These effects of LSF inhibition, mitotic arrest and induction of apoptosis by FQI1s provide multiple avenues by which these inhibitors eliminate HCC cells. LSF inhibitors might be highly potent and effective therapeutics for HCC either alone or in combination with currently existing therapies.

  11. Expression profiles of a MhCTR1 gene in relation to banana fruit ripening.

    PubMed

    Hu, Huei-Lin; Do, Yi-Yin; Huang, Pung-Ling

    2012-07-01

    The banana (Musa spp.) is a typical climacteric fruit of high economic importance. The development of bananas from maturing to ripening is characterized by increased ethylene production accompanied by a respiration burst. To elucidate the signal transduction pathway involved in the ethylene regulation of banana ripening, a gene homologous to Arabidopsis CTR1 (constitutive triple response 1) was isolated from Musa spp. (Hsien Jin Chiao, AAA group) and designated as MhCTR1. MhCTR1 spans 11.5 kilobases and consists of 15 exons and 14 introns with consensus GT-AG nucleotides situated at their boundaries. MhCTR1 encodes a polypeptide of 805 amino acid residues with a calculated molecular weight of 88.6 kDa. The deduced amino acid sequence of MhCTR1 demonstrates 55%, 56% and 55% homology to AtCTR1, RhCTR1, and LeCTR1, respectively. MhCTR1 is expressed mostly in the mature green pulp and root organs. During fruit development MhCTR1 expression increases just before ethylene production rises. Moreover, MhCTR1 expression was detected mainly in the pulps at ripening stage 3, and correlated with the onset of peel yellowing, while MhCTR1 was constitutively expressed in the peels. MhCTR1 expression could be induced by ethylene treatment (0.01 μL L(-1)), and MhCTR1 expression decreased in both peel and pulp 24 h after treatment. Overall, changes observed in MhCTR1 expression in the pulp closely related to the regulation of the banana ripening process. Copyright © 2012. Published by Elsevier Masson SAS.

  12. Effect of LED irradiation on the ripening and nutritional quality of postharvest banana fruit.

    PubMed

    Huang, Jen-Yi; Xu, Fengying; Zhou, Weibiao

    2018-04-24

    With the ability to tailor wavelengths necessary to the photosynthetically active radiation spectrum of plant pigments, light-emitting diodes (LEDs) offer vast possibilities in horticultural lighting. The influence of LED light irradiation on major postharvest features of banana was investigated. Mature green bananas were treated daily with selected blue (464-474 nm), green (515-525 nm) and red (617-627 nm) LED lights for 8 days, and compared with non-illuminated control. The positive effect of LED lighting on the acceleration of ripening in bananas was greatest for blue, followed by red and green. Under the irradiation of LED lights, faster peel de-greening and flesh softening, and increased ethylene production and respiration rate in bananas were observed during storage. Furthermore, the accumulations of ascorbic acid, total phenols, and total sugars in banana fruit were enhanced by LED light exposure. LED light treatment can induce the ripening of bananas and improve their quality and nutrition potential. These findings might provide new chemical-free strategies to shorten the time to ripen banana after harvest by using LED light source. This article is protected by copyright. All rights reserved.

  13. Effects of different strains Penicillium nalgiovense in the Nalžovy cheese during ripening.

    PubMed

    Mrázek, Josef; Pachlová, Vendula; Buňka, František; Černíková, Michaela; Dráb, Vladimír; Bejblová, Martina; Staněk, Karel; Buňková, Leona

    2016-05-01

    The present study examined the influence of eight selected strains of Penicillium nalgiovense as secondary culture on the ripening of soft mould cheese. Ion-exchange chromatography, SDS-PAGE and sensory analysis were used for comparison of P. nalgiovense with common secondary mould culture (Penicillium camemberti). Proteolytic activity was more intensive in the case of the strains of P. nalgiovense in comparison with the control strains of P. camemberti. Some differences in the proportion and amount of free amino acids were observed between the strains of P. nalgiovense during ripening. Lower proteolytic activity of P. nalgiovense was more desirable for sensory properties of cheese, especially in case sample N2. This study provides information on the usability of selected strains of P. nalgiovense that could support the development of sensory properties of mould-ripened soft cheese. The cheese with the culture N2 had the best flavour after 35 days of storage. Moreover, this strain showed the lowest bitterness in comparison with the other used P. nalgiovense cultures. The intensity of bitterness of sample N2 was comparable to that of the control sample with P. camemberti. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  14. Dynamics of bacterial communities during the ripening process of different Croatian cheese types derived from raw ewe's milk cheeses.

    PubMed

    Fuka, Mirna Mrkonjić; Wallisch, Stefanie; Engel, Marion; Welzl, Gerhard; Havranek, Jasmina; Schloter, Michael

    2013-01-01

    Microbial communities play an important role in cheese ripening and determine the flavor and taste of different cheese types to a large extent. However, under adverse conditions human pathogens may colonize cheese samples during ripening and may thus cause severe outbreaks of diarrhoea and other diseases. Therefore in the present study we investigated the bacterial community structure of three raw ewe's milk cheese types, which are produced without the application of starter cultures during ripening from two production sites based on fingerprinting in combination with next generation sequencing of 16S rRNA gene amplicons. Overall a surprisingly high diversity was found in the analyzed samples and overall up to 213 OTU97 could be assigned. 20 of the major OTUs were present in all samples and include mostly lactic acid bacteria (LAB), mainly Lactococcus, and Enterococcus species. Abundance and diversity of these genera differed to a large extent between the 3 investigated cheese types and in response to the ripening process. Also a large number of non LAB genera could be identified based on phylogenetic alignments including mainly Enterobacteriaceae and Staphylococcacae. Some species belonging to these two families could be clearly assigned to species which are known as potential human pathogens like Staphylococcus saprophyticus or Salmonella spp. However, during cheese ripening their abundance was reduced. The bacterial genera, namely Lactobacillus, Streptococcus, Leuconostoc, Bifidobacterium, Brevibacterium, Corynebacterium, Clostridium, Staphylococcus, Thermoanerobacterium, E. coli, Hafnia, Pseudomonas, Janthinobacterium, Petrotoga, Kosmotoga, Megasphaera, Macrococcus, Mannheimia, Aerococcus, Vagococcus, Weissella and Pediococcus were identified at a relative low level and only in selected samples. Overall the microbial composition of the used milk and the management of the production units determined the bacterial community composition for all cheese types to a

  15. Dynamics of Bacterial Communities during the Ripening Process of Different Croatian Cheese Types Derived from Raw Ewe's Milk Cheeses

    PubMed Central

    Fuka, Mirna Mrkonjić; Wallisch, Stefanie; Engel, Marion; Welzl, Gerhard; Havranek, Jasmina; Schloter, Michael

    2013-01-01

    Microbial communities play an important role in cheese ripening and determine the flavor and taste of different cheese types to a large extent. However, under adverse conditions human pathogens may colonize cheese samples during ripening and may thus cause severe outbreaks of diarrhoea and other diseases. Therefore in the present study we investigated the bacterial community structure of three raw ewe's milk cheese types, which are produced without the application of starter cultures during ripening from two production sites based on fingerprinting in combination with next generation sequencing of 16S rRNA gene amplicons. Overall a surprisingly high diversity was found in the analyzed samples and overall up to 213 OTU97 could be assigned. 20 of the major OTUs were present in all samples and include mostly lactic acid bacteria (LAB), mainly Lactococcus, and Enterococcus species. Abundance and diversity of these genera differed to a large extent between the 3 investigated cheese types and in response to the ripening process. Also a large number of non LAB genera could be identified based on phylogenetic alignments including mainly Enterobacteriaceae and Staphylococcacae. Some species belonging to these two families could be clearly assigned to species which are known as potential human pathogens like Staphylococcus saprophyticus or Salmonella spp. However, during cheese ripening their abundance was reduced. The bacterial genera, namely Lactobacillus, Streptococcus, Leuconostoc, Bifidobacterium, Brevibacterium, Corynebacterium, Clostridium, Staphylococcus, Thermoanerobacterium, E. coli, Hafnia, Pseudomonas, Janthinobacterium, Petrotoga, Kosmotoga, Megasphaera, Macrococcus, Mannheimia, Aerococcus, Vagococcus, Weissella and Pediococcus were identified at a relative low level and only in selected samples. Overall the microbial composition of the used milk and the management of the production units determined the bacterial community composition for all cheese types to a

  16. Metabolomic analysis of avocado fruits by GC-APCI-TOF MS: effects of ripening degrees and fruit varieties.

    PubMed

    Hurtado-Fernández, E; Pacchiarotta, T; Mayboroda, O A; Fernández-Gutiérrez, A; Carrasco-Pancorbo, A

    2015-01-01

    In order to investigate avocado fruit ripening, nontargeted GC-APCI-TOF MS metabolic profiling analyses were carried out. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used to explore the metabolic profiles from fruit samples of 13 varieties at two different ripening degrees. Mannoheptulose; pentadecylfuran; aspartic, malic, stearic, citric and pantothenic acids; mannitol; and β-sitosterol were some of the metabolites found as more influential for the PLS-DA model. The similarities among genetically related samples (putative mutants of "Hass") and their metabolic differences from the rest of the varieties under study have also been evaluated. The achieved results reveal new insights into avocado fruit composition and metabolite changes, demonstrating therefore the value of metabolomics as a functional genomics tool in characterizing the mechanism of fruit ripening development, a key developmental stage in most economically important fruit crops.

  17. Small-Molecule Inhibitors of the SOX18 Transcription Factor.

    PubMed

    Fontaine, Frank; Overman, Jeroen; Moustaqil, Mehdi; Mamidyala, Sreeman; Salim, Angela; Narasimhan, Kamesh; Prokoph, Nina; Robertson, Avril A B; Lua, Linda; Alexandrov, Kirill; Koopman, Peter; Capon, Robert J; Sierecki, Emma; Gambin, Yann; Jauch, Ralf; Cooper, Matthew A; Zuegg, Johannes; Francois, Mathias

    2017-03-16

    Pharmacological modulation of transcription factors (TFs) has only met little success over the past four decades. This is mostly due to standard drug discovery approaches centered on blocking protein/DNA binding or interfering with post-translational modifications. Recent advances in the field of TF biology have revealed a central role of protein-protein interaction in their mode of action. In an attempt to modulate the activity of SOX18 TF, a known regulator of vascular growth in development and disease, we screened a marine extract library for potential small-molecule inhibitors. We identified two compounds, which inspired a series of synthetic SOX18 inhibitors, able to interfere with the SOX18 HMG DNA-binding domain, and to disrupt HMG-dependent protein-protein interaction with RBPJ. These compounds also perturbed SOX18 transcriptional activity in a cell-based reporter gene system. This approach may prove useful in developing a new class of anti-angiogenic compounds based on the inhibition of TF activity. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Climacteric ripening of apple fruit is regulated by transcriptional circuits stimulated by cross-talks between ethylene and auxin.

    PubMed

    Busatto, Nicola; Tadiello, Alice; Trainotti, Livio; Costa, Fabrizio

    2017-01-02

    Apple is a fleshy fruit distinguished by a climacteric type of ripening, since most of the relevant physiological changes are triggered and governed by the action of ethylene. After its production, this hormone is perceived by a series of receptors to regulate, through a signaling cascade, downstream ethylene related genes. The possibility to control the effect of ethylene opened new horizons to the improvement of the postharvest fruit quality. To this end, 1-methylcyclopropene (1-MCP), an ethylene antagonist, is routinely used to modulate the ripening progression increasing storage life. In a recent work published in The Plant Journal, the whole transcriptome variation throughout fruit development and ripening, with the adjunct comparison between normal and impaired postharvest ripening, has been illustrated. In particular, besides the expected downregulation of ethylene-regulated genes, we shed light on a regulatory circuit leading to de-repressing the expression of a specific set of genes following 1-MCP treatment, such as AUX/IAA, NAC and MADS. These findings suggested the existence of a possible ethylene/auxin cross-talk in apple, regulated by a transcriptional circuit stimulated by the interference at the ethylene receptor level.

  19. Factor Xa inhibitors versus vitamin K antagonists for preventing cerebral or systemic embolism in patients with atrial fibrillation.

    PubMed

    Bruins Slot, Karsten Mh; Berge, Eivind

    2018-03-06

    Factor Xa inhibitors and vitamin K antagonists (VKAs) are now recommended in treatment guidelines for preventing stroke and systemic embolic events in people with atrial fibrillation (AF). This is an update of a Cochrane review previously published in 2013. To assess the effectiveness and safety of treatment with factor Xa inhibitors versus VKAs for preventing cerebral or systemic embolic events in people with AF. We searched the trials registers of the Cochrane Stroke Group and the Cochrane Heart Group (September 2016), the Cochrane Central Register of Controlled Trials (CENTRAL) (August 2017), MEDLINE (1950 to April 2017), and Embase (1980 to April 2017). We also contacted pharmaceutical companies, authors and sponsors of relevant published trials. We used outcome data from marketing authorisation applications of apixaban, edoxaban and rivaroxaban that were submitted to regulatory authorities in Europe and the USA. We included randomised controlled trials (RCTs) that directly compared the effects of long-term treatment (lasting more than four weeks) with factor Xa inhibitors versus VKAs for preventing cerebral and systemic embolism in people with AF. The primary efficacy outcome was the composite endpoint of all strokes and systemic embolic events. Two review authors independently extracted data, and assessed the quality of the trials and the risk of bias. We calculated a weighted estimate of the typical treatment effect across trials using the odds ratio (OR) with 95% confidence interval (CI) by means of a fixed-effect model. In case of moderate or high heterogeneity of treatment effects, we used a random-effects model to compare the overall treatment effects. We also performed a pre-specified sensitivity analysis excluding any open-label studies. We included data from 67,688 participants randomised into 13 RCTs. The included trials directly compared dose-adjusted warfarin with either apixaban, betrixaban, darexaban, edoxaban, idraparinux, idrabiotaparinux, or

  20. Comparative transcriptome analysis of unripe and mid-ripe fruit of Mangifera indica (var. “Dashehari”) unravels ripening associated genes

    PubMed Central

    Srivastava, Smriti; Singh, Rajesh K.; Pathak, Garima; Goel, Ridhi; Asif, Mehar Hasan; Sane, Aniruddha P.; Sane, Vidhu A.

    2016-01-01

    Ripening in mango is under a complex control of ethylene. In an effort to understand the complex spatio-temporal control of ripening we have made use of a popular N. Indian variety “Dashehari” This variety ripens from the stone inside towards the peel outside and forms jelly in the pulp in ripe fruits. Through a combination of 454 and Illumina sequencing, a transcriptomic analysis of gene expression from unripe and midripe stages have been performed in triplicates. Overall 74,312 unique transcripts with ≥1 FPKM were obtained. The transcripts related to 127 pathways were identified in “Dashehari” mango transcriptome by the KEGG analysis. These pathways ranged from detoxification, ethylene biosynthesis, carbon metabolism and aromatic amino acid degradation. The transcriptome study reveals differences not only in expression of softening associated genes but also those that govern ethylene biosynthesis and other nutritional characteristics. This study could help to develop ripening related markers for selective breeding to reduce the problems of excess jelly formation during softening in the “Dashehari” variety. PMID:27586495

  1. A SEPALLATA gene is involved in the development and ripening of strawberry (Fragaria x ananassa Duch.) fruit, a non-climacteric tissue

    USDA-ARS?s Scientific Manuscript database

    Climacteric and non-climacteric fruits have traditionally been viewed as representing two distinct programs of ripening associated with differential respiration and ethylene hormone effects. In climacteric fruits, such as tomato and banana, the ripening process is marked by increased respiration and...

  2. Plasmin-dependent proteolysis of Tissue Factor Pathway Inhibitor in a mouse model of endotoxemia

    PubMed Central

    Lupu, Cristina; Herlea, Oana; Tang, Haiwang; Lijnen, Roger H.; Lupu, Florea

    2012-01-01

    Summary Background Development of a procoagulant state in sepsis, due to aberrant expression of tissue factor (TF) and sharp decrease of its major inhibitor tissue factor pathway inhibitor (TFPI), could lead to microthrombotic organ failure. The mechanism for the decline of TFPI activity in the lung could involve plasmin-mediated cleavage of the inhibitor. Objective To investigate the effect of plasmin generation on lung-associated TFPI activity, in normal conditions and during infusion of endotoxin (LPS) in mice. Methods Plasmin generation and TFPI activity were assayed in the lungs of mice deficient of tissue-type plasminogen activator (t-PA) or plasminogen (Plg), at 2-hrs after LPS or saline injection. Results The sharp loss of lung-associated TFPI activity at 2-hrs post LPS paralleled the abrupt increase of plasmin generation. TFPI activity was significantly retained in both t-PA-/- and Plg-/- mice, which are unable to generate plasmin. Conclusion The increased plasmin generation during the early stages of sepsis could cleave/inactivate TFPI and thus lead to thrombotic complications. PMID:23106863

  3. Portable Raman spectroscopy for an in-situ monitoring the ripening of tomato (Solanum lycopersicum) fruits

    NASA Astrophysics Data System (ADS)

    Trebolazabala, Josu; Maguregui, Maite; Morillas, Héctor; de Diego, Alberto; Madariaga, Juan Manuel

    2017-06-01

    Ripening is one of the most important transformations that fruits and vegetables suffer, from an unripe to a ripe stage. In this study, it was followed up and analyzed the variations in the composition of tomato fruits at different ripening stages (green or unripe, orange or middle ripe, red or ripe and brown or overripe). The results obtained from the Raman measurements carried out showed a change in the composition of tomato fruits in the transit from green to brown. The analysis confirmed an increase of carotenoids from an unripe to a ripe stage of these fruits, being lycopene the characteristic carotenoid of the optimum ripe stage. The presence of chlorophyll and cuticular waxes decrease from the unripe to the ripe stage. Moreover, the relative intensity of phytofluene, a transition compound in the carotenoid biosynthetic pathway, is higher in the orange or middle ripening stage. The results obtained in-situ, without cutting and handling the tomato fruits, by means of a portable Raman spectrometer offered the same information that can be achieved using a more expensive and sophisticated confocal Raman microscope.

  4. Modeling the growth of Listeria monocytogenes in mold-ripened cheeses.

    PubMed

    Lobacz, Adriana; Kowalik, Jaroslaw; Tarczynska, Anna

    2013-06-01

    This study presents possible applications of predictive microbiology to model the safety of mold-ripened cheeses with respect to bacteria of the species Listeria monocytogenes during (1) the ripening of Camembert cheese, (2) cold storage of Camembert cheese at temperatures ranging from 3 to 15°C, and (3) cold storage of blue cheese at temperatures ranging from 3 to 15°C. The primary models used in this study, such as the Baranyi model and modified Gompertz function, were fitted to growth curves. The Baranyi model yielded the most accurate goodness of fit and the growth rates generated by this model were used for secondary modeling (Ratkowsky simple square root and polynomial models). The polynomial model more accurately predicted the influence of temperature on the growth rate, reaching the adjusted coefficients of multiple determination 0.97 and 0.92 for Camembert and blue cheese, respectively. The observed growth rates of L. monocytogenes in mold-ripened cheeses were compared with simulations run with the Pathogen Modeling Program (PMP 7.0, USDA, Wyndmoor, PA) and ComBase Predictor (Institute of Food Research, Norwich, UK). However, the latter predictions proved to be consistently overestimated and contained a significant error level. In addition, a validation process using independent data generated in dairy products from the ComBase database (www.combase.cc) was performed. In conclusion, it was found that L. monocytogenes grows much faster in Camembert than in blue cheese. Both the Baranyi and Gompertz models described this phenomenon accurately, although the Baranyi model contained a smaller error. Secondary modeling and further validation of the generated models highlighted the issue of usability and applicability of predictive models in the food processing industry by elaborating models targeted at a specific product or a group of similar products. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  5. Different Preclimacteric Events in Apple Cultivars with Modified Ripening Physiology

    PubMed Central

    Singh, Vikram; Weksler, Asya; Friedman, Haya

    2017-01-01

    “Anna” is an early season apple cultivar exhibiting a fast softening and juiciness loss during storage, in comparison to two mid-late season cultivars “Galaxy” and “GD.” The poor storage capacity of “Anna” was correlated with high lipid oxidation-related autoluminescence, high respiration and ethylene production rates, associated with high expression of MdACO1, 2, 4, 7, and MdACS1. All cultivars at harvest responded to exogenous ethylene by enhancing ethylene production, typical of system-II. The contribution of pre-climacteric events to the poor storage capacity of “Anna” was examined by comparing respiration and ethylene production rates, response to exogenous ethylene, expression of genes responsible for ethylene biosynthesis and response, and developmental regulators in the three cultivars throughout fruit development. In contrast to the “Galaxy” and “GD,” “Anna” showed higher ethylene production and respiration rates during fruit development, and exhibited auto-stimulatory (system II-like) effect in response to exogenous ethylene. The higher ethylene production rate in “Anna” was correlated with higher expression of ethylene biosynthesis genes, MdACS3a MdACO2, 4, and 7 during early fruit development. The expression of negative regulators of ripening (AP2/ERF) and ethylene response pathway, (MdETR1,2 and MdCTR1) was lower in “Anna” in comparison to the other two cultivars throughout development and ripening. Similar pattern of gene expression was found for SQUAMOSA promoter binding protein (SBP)-box genes, including MdCNR and for MdFUL. Taken together, this study provides new understanding on pre-climacteric events in “Anna” that might affect its ripening behavior and physiology following storage. PMID:28928755

  6. Simultaneous expression of tissue factor and tissue factor pathway inhibitor by human monocytes. A potential mechanism for localized control of blood coagulation

    PubMed Central

    1994-01-01

    Cells of monocytic lineage can initiate extravascular fibrin deposition via expression of blood coagulation mediators. This report is about experiments on three mechanisms with the potential to modulate monocyte- initiated coagulation. Monocyte procoagulant activity was examined as a function of lipid cofactor, protein cofactor, and specific inhibitor expression during short-term culture in vitro. Lipid cofactor activity was measured as the initial rate of factor X activation by intrinsic- pathway components, the assembly of which depends on this cofactor. Lipid cofactor activity levels changed by < 30% during 48-h culture. Protein cofactor, i.e., tissue factor (TF) antigen was measured by enzyme immunoassay. It increased from 461 pg/ml to a maximum value of 3,550 pg/ml at 24 h and remained at 70% of this value. Specific TF activity, measured as factor VII-dependent factor X activation rate, decreased from 54 to 18 nM FXa/min between 24 and 48 h. TF activity did not correlate well with either lipid cofactor or TF protein levels. In contrast, the decrease in TF activity coincided in time with maximal expression of tissue factor pathway inhibitor (TFPI) mRNA, which was determined using reverse transcriptase polymerase chain reaction (RT- PCR), and with maximal TFPI protein levels measured by immunoassay. The number of mRNA copies coding for TFPI and TF in freshly isolated blood monocytes were 46 and 20 copies/cells, respectively. These values increased to 220 and 63 copies/cell during short-term cell culture in the presence of endotoxin. Results demonstrate concomitant expression by monocytes of genes coding for both the essential protein cofactor and the specific inhibitor of the extrinsic coagulation pathway. Together with functional and antigenic analyses, they also imply that the initiation of blood clotting by extravascular monocyte/macrophages can be modulated locally by TFPI independently of plasma sources of the inhibitor. PMID:8195712

  7. Mango (Mangifera indica L.) cv. Kent fruit mesocarp de novo transcriptome assembly identifies gene families important for ripening.

    PubMed

    Dautt-Castro, Mitzuko; Ochoa-Leyva, Adrian; Contreras-Vergara, Carmen A; Pacheco-Sanchez, Magda A; Casas-Flores, Sergio; Sanchez-Flores, Alejandro; Kuhn, David N; Islas-Osuna, Maria A

    2015-01-01

    Fruit ripening is a physiological and biochemical process genetically programmed to regulate fruit quality parameters like firmness, flavor, odor and color, as well as production of ethylene in climacteric fruit. In this study, a transcriptomic analysis of mango (Mangifera indica L.) mesocarp cv. "Kent" was done to identify key genes associated with fruit ripening. Using the Illumina sequencing platform, 67,682,269 clean reads were obtained and a transcriptome of 4.8 Gb. A total of 33,142 coding sequences were predicted and after functional annotation, 25,154 protein sequences were assigned with a product according to Swiss-Prot database and 32,560 according to non-redundant database. Differential expression analysis identified 2,306 genes with significant differences in expression between mature-green and ripe mango [1,178 up-regulated and 1,128 down-regulated (FDR ≤ 0.05)]. The expression of 10 genes evaluated by both qRT-PCR and RNA-seq data was highly correlated (R = 0.97), validating the differential expression data from RNA-seq alone. Gene Ontology enrichment analysis, showed significantly represented terms associated to fruit ripening like "cell wall," "carbohydrate catabolic process" and "starch and sucrose metabolic process" among others. Mango genes were assigned to 327 metabolic pathways according to Kyoto Encyclopedia of Genes and Genomes database, among them those involved in fruit ripening such as plant hormone signal transduction, starch and sucrose metabolism, galactose metabolism, terpenoid backbone, and carotenoid biosynthesis. This study provides a mango transcriptome that will be very helpful to identify genes for expression studies in early and late flowering mangos during fruit ripening.

  8. Pharmacophore selection and redesign of non-nucleotide inhibitors of anthrax edema factor.

    PubMed

    Schein, Catherine H; Chen, Deliang; Ma, Lili; Kanalas, John J; Gao, Jian; Jimenez, Maria Estrella; Sower, Laurie E; Walter, Mary A; Gilbertson, Scott R; Peterson, Johnny W

    2012-11-08

    Antibiotic treatment may fail to protect individuals, if not started early enough, after infection with Bacillus anthracis, due to the continuing activity of toxins that the bacterium produces. Stable and easily stored inhibitors of the edema factor toxin (EF), an adenylyl cyclase, could save lives in the event of an outbreak, due to natural causes or a bioweapon attack. The toxin's basic activity is to convert ATP to cAMP, and it is thus in principle a simple phosphatase, which means that many mammalian enzymes, including intracellular adenylcyclases, may have a similar activity. While nucleotide based inhibitors, similar to its natural substrate, ATP, were identified early, these compounds had low activity and specificity for EF. We used a combined structural and computational approach to choose small organic molecules in large, web-based compound libraries that would, based on docking scores, bind to residues within the substrate binding pocket of EF. A family of fluorenone-based inhibitors was identified that inhibited the release of cAMP from cells treated with EF. The lead inhibitor was also shown to inhibit the diarrhea caused by enterotoxigenic E. coli (ETEC) in a murine model, perhaps by serving as a quorum sensor. These inhibitors are now being tested for their ability to inhibit Anthrax infection in animal models and may have use against other pathogens that produce toxins similar to EF, such as Bordetella pertussis or Vibrio cholera.

  9. Effect of CRC::etr1-1 transgene expression on ethylene production, sex expression, fruit set and fruit ripening in transgenic melon (Cucumis melo L.).

    PubMed

    Switzenberg, Jessica A; Beaudry, Randy M; Grumet, Rebecca

    2015-06-01

    Ethylene is a key factor regulating sex expression in cucurbits. Commercial melons (Cucumis melo L.) are typically andromonoecious, producing male and bisexual flowers. Our prior greenhouse studies of transgenic melon plants expressing the dominant negative ethylene perception mutant gene, etr1-1, under control of the carpel- and nectary-primordia targeted CRAB'S CLAW (CRC) promoter showed increased number and earlier appearance of carpel-bearing flowers. To further investigate this phenomenon which could be potentially useful for earlier fruit production, we observed CRC::etr1-1 plants in the field for sex expression, fruit set, fruit development, and ripening. CRC::etr1-1 melon plants showed increased number of carpel-bearing open flowers on the main stem and earlier onset by 7-10 nodes. Additional phenotypes observed in the greenhouse and field were conversion of approximately 50% of bisexual buds to female, and elongated ovaries and fruits. Earlier and greater fruit set occurred on the transgenic plants. However, CRC::etr1-1 plants had greater abscission of young fruit, and smaller fruit, so that final yield (kg/plot) was equivalent to wild type. Earlier fruit set in line M5 was accompanied by earlier appearance of ripe fruit. Fruit from line M15 frequently did not exhibit external ripening processes of rind color change and abscission, but when cut open, the majority showed a ripe or overripe interior accompanied by elevated internal ethylene. The non-ripening external phenotype in M15 fruit corresponded with elevated etr1-1 transgene expression in the exocarp. These results provide insight into the role of ethylene perception in carpel-bearing flower production, fruit set, and ripening.

  10. Comparative Analysis of Fruit Ripening-Related miRNAs and Their Targets in Blueberry Using Small RNA and Degradome Sequencing

    PubMed Central

    Hou, Yanming; Zhai, Lulu; Li, Xuyan; Xue, Yu; Wang, Jingjing; Yang, Pengjie; Cao, Chunmei; Li, Hongxue; Cui, Yuhai; Bian, Shaomin

    2017-01-01

    MicroRNAs (miRNAs) play vital roles in the regulation of fruit development and ripening. Blueberry is an important small berry fruit crop with economical and nutritional value. However, nothing is known about the miRNAs and their targets involved in blueberry fruit ripening. In this study, using high-throughput sequencing of small RNAs, 84 known miRNAs belonging to 28 families and 16 novel miRNAs were identified in white fruit (WF) and blue fruit (BF) libraries, which represent fruit ripening onset and in progress, respectively. Among them, 41 miRNAs were shown to be differentially expressed during fruit maturation, and 16 miRNAs representing 16 families were further chosen to validate the sRNA sequencing data by stem-loop qRT-PCR. Meanwhile, 178 targets were identified for 41 known and 7 novel miRNAs in WF and BF libraries using degradome sequencing, and targets of miR160 were validated using RLM-RACE (RNA Ligase-Mediated (RLM)-Rapid Amplification of cDNA Ends) approach. Moreover, the expression patterns of 6 miRNAs and their targets were examined during fruit development and ripening. Finally, integrative analysis of miRNAs and their targets revealed a complex miRNA-mRNA regulatory network involving a wide variety of biological processes. The findings will facilitate future investigations of the miRNA-mediated mechanisms that regulate fruit development and ripening in blueberry. PMID:29257112

  11. Evolution of total and individual capsaicinoids in peppers during ripening of the Cayenne pepper plant (Capsicum annuum L.).

    PubMed

    Barbero, Gerardo F; Ruiz, Aurora G; Liazid, Ali; Palma, Miguel; Vera, Jesús C; Barroso, Carmelo G

    2014-06-15

    The evolution of total capsaicinoids and the individual contents of the five major capsaicinoids: nordihydrocapsaicin, capsaicin, dihydrocapsaicin, homocapsaicin and homodihydrocapsaicin present in the Cayenne pepper (Capsicum annuum L.), during fruit ripening, has been established. Capsaicinoids begin to accumulate gradually in the peppers from the beginning of its development up to a maximum concentration (1,789 μmol/Kg FW). From this time there is initially a sharp decrease in the total capsaicinoid content (32%), followed by a gradual decrease until day 80 of ripening. The two major capsaicinoids present in the Cayenne pepper are capsaicin and dihydrocapsaicin, which represent between 79% and 90%, respectively, of total capsaicinoids depending on fruit ripening. The relative content of capsaicin differs from the evolution of the other four capsaicinoids studied. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Transcriptome profiling analysis of cultivar-specific apple fruit ripening and texture attributes

    USDA-ARS?s Scientific Manuscript database

    Molecular events regulating cultivar-specific apple fruit ripening and sensory quality are largely unknown. Such knowledge is essential for genomic-assisted apple breeding and postharvest quality management. In this study, transcriptome profile analysis, scanning electron microscopic examination an...

  13. Development of Inhibitors of the PAS-B Domain of the HIF-2α Transcription Factor

    PubMed Central

    Rogers, Jamie L.; Bayeh, Liela; Scheuermann, Thomas H.; Longgood, Jamie; Key, Jason; Naidoo, Jacinth; Melito, Lisa; Shokri, Cameron; Frantz, Doug E.; Bruick, Richard K.; Gardner, Kevin H.; MacMillan, John B.; Tambar, Uttam K.

    2013-01-01

    Hypoxia Inducible Factors (HIFs) are heterodimeric transcription factors induced in a variety of pathophysiological settings, including cancer. We describe the first detailed structure-activity-relationship study of small molecules designed to inhibit HIF-2α–ARNT heterodimerization by binding an internal cavity of the HIF-2α PAS-B domain. Through a series of biophysical characterizations of inhibitor/protein interactions (NMR and X-ray crystallography), we have established the structural requirements for artificial inhibitors of the HIF-2α–ARNT PAS-B interaction. These results may serve as a foundation for discovering therapeutic agents that function by a novel mode of action. PMID:23363003

  14. Monitoring the ripening process of Cheddar cheese based on hydrophilic component profiling using gas chromatography-mass spectrometry.

    PubMed

    Ochi, H; Sakai, Y; Koishihara, H; Abe, F; Bamba, T; Fukusaki, E

    2013-01-01

    We proposed an application methodology that combines metabolic profiling with multiple appropriate multivariate analyses and verified it on the industrial scale of the ripening process of Cheddar cheese to make practical use of hydrophilic low-molecular-weight compound profiling using gas chromatography-mass spectrometry to design optimal conditions and quality monitoring of the cheese ripening process. Principal components analysis provided an overview of the effect of sodium chloride content and kind of lactic acid bacteria starter on the metabolic profile in the ripening process of Cheddar cheese and orthogonal partial least squares-discriminant analysis unveiled the difference in characteristic metabolites. When the sodium chloride contents were different (1.6 and 0.2%) but the same lactic acid bacteria starter was used, the 2 cheeses were classified by orthogonal partial least squares-discriminant analysis from their metabolic profiles, but were not given perfect discrimination. Not much difference existed in the metabolic profile between the 2 cheeses. Compounds including lactose, galactose, lactic acid, 4-aminobutyric acid, and phosphate were identified as contents that differed between the 2 cheeses. On the other hand, in the case of the same salt content of 1.6%, but different kinds of lactic acid bacteria starter, an excellent distinctive discrimination model was obtained, which showed that the difference of lactic acid bacteria starter caused an obvious difference in metabolic profiles. Compounds including lactic acid, lactose, urea, 4-aminobutyric acid, galactose, phosphate, proline, isoleucine, glycine, alanine, lysine, leucine, valine, and pyroglutamic acid were identified as contents that differed between the 2 cheeses. Then, a good sensory prediction model for "rich flavor," which was defined as "thick and rich, including umami taste and soy sauce-like flavor," was constructed based on the metabolic profile during ripening using partial least

  15. The common transcriptional subnetworks of the grape berry skin in the late stages of ripening.

    PubMed

    Ghan, Ryan; Petereit, Juli; Tillett, Richard L; Schlauch, Karen A; Toubiana, David; Fait, Aaron; Cramer, Grant R

    2017-05-30

    Wine grapes are important economically in many countries around the world. Defining the optimum time for grape harvest is a major challenge to the grower and winemaker. Berry skins are an important source of flavor, color and other quality traits in the ripening stage. Senescent-like processes such as chloroplast disorganization and cell death characterize the late ripening stage. To better understand the molecular and physiological processes involved in the late stages of berry ripening, RNA-seq analysis of the skins of seven wine grape cultivars (Cabernet Franc, Cabernet Sauvignon, Merlot, Pinot Noir, Chardonnay, Sauvignon Blanc and Semillon) was performed. RNA-seq analysis identified approximately 2000 common differentially expressed genes for all seven cultivars across four different berry sugar levels (20 to 26 °Brix). Network analyses, both a posteriori (standard) and a priori (gene co-expression network analysis), were used to elucidate transcriptional subnetworks and hub genes associated with traits in the berry skins of the late stages of berry ripening. These independent approaches revealed genes involved in photosynthesis, catabolism, and nucleotide metabolism. The transcript abundance of most photosynthetic genes declined with increasing sugar levels in the berries. The transcript abundance of other processes increased such as nucleic acid metabolism, chromosome organization and lipid catabolism. Weighted gene co-expression network analysis (WGCNA) identified 64 gene modules that were organized into 12 subnetworks of three modules or more and six higher order gene subnetworks. Some gene subnetworks were highly correlated with sugar levels and some subnetworks were highly enriched in the chloroplast and nucleus. The petal R package was utilized independently to construct a true small-world and scale-free complex gene co-expression network model. A subnetwork of 216 genes with the highest connectivity was elucidated, consistent with the module results

  16. Dynamics in the concentrations of health-promoting compounds: lupeol, mangiferin and different phenolic acids during postharvest ripening of mango fruit.

    PubMed

    Vithana, Mekhala Dk; Singh, Zora; Johnson, Stuart K

    2018-03-01

    Mango fruit (Mangifera indica L.) is renowned for its pleasant taste and as a rich source of health beneficial compounds. The aim of this study was to investigate the changes in concentrations of health-promoting compounds, namely ascorbic acid, carotenoids, antioxidants, lupeol, mangiferin, total phenols and individual phenolic acids, as well as ethylene production and respiration rates during climacteric ripening in 'Kensington Pride' and 'R2E2' mango fruit. The climacteric ethylene and respiration peaks were noted on the third day of the fruit ripening period. The concentrations of total carotenoids in the pulp, total antioxidants in both pulp and peel, and total phenols of the peel, lupeol and mangiferin were significantly elevated, whereas the concentration of ascorbic acid declined during post-climacteric ripening. Gallic, chlorogenic and vanillic acids were identified as the major phenolic acids in both pulp and peel of 'Kensington Pride' and 'R2E2' mangoes. The concentrations of phenolic acids (gallic, chlorogenic, vanillic, ferulic and caffeic acids) also increased during the post-climacteric phase. The concentrations of all phenolic compounds were several-fold higher in the peel than pulp. Mangoes at post-climacteric ripening phase offer the highest concentrations of health-promoting compounds. Peel, at this stage of fruit ripening, could be exploited as a good source for extraction of these compounds. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  17. Influence of starters on chemical, biochemical, and sensory changes in Turkish White-brined cheese during ripening.

    PubMed

    Hayaloglu, A A; Guven, M; Fox, P F; McSweeney, P L H

    2005-10-01

    Turkish White-brined cheese was manufactured using Lactococcus strains (Lactococcus lactis ssp. lactis NCDO763 plus L. lactis ssp. cremoris SK11 and L. lactis ssp. lactis UC317 plus L. lactis ssp. cremoris HP) or without a starter culture, and ripened for 90 d. It was found that the use of starters significantly influenced the physical, chemical, biochemical, and sensory properties of the cheeses. Chemical composition, pH, and sensory properties of cheeses made with starter were not affected by the different starter bacteria. The levels of soluble nitrogen fractions and urea-PAGE of the pH 4.6-insoluble fractions were found to be significantly different at various stages of ripening. Urea-PAGE patterns of the pH 4.6-insoluble fractions of the cheeses showed that considerable degradation of alpha(s1)-casein occurred and that beta-casein was more resistant to hydrolysis. The use of a starter culture significantly influenced the levels of 12% trichloroacetic acid-soluble nitrogen, 5% phosphotungstic acid-soluble nitrogen, free amino acids, total free fatty acids, and the peptide profiles (reverse phase-HPLC) of 70% (vol/vol) ethanol-soluble and insoluble fractions of the pH 4.6-soluble fraction of the cheeses. The levels of peptides in the cheeses increased during the ripening period. Principal component and hierarchical cluster analyses of electrophoretic and chromatographic results indicated that the cheeses were significantly different in terms of their peptide profiles and they were grouped based on the use and type of starter and stage of ripening. Levels of free amino acid in the cheeses differed; Leu, Glu, Phe, Lys, and Val were the most abundant amino acids. Nitrogen fractions, total free amino acids, total free fatty acids, and the levels of peptides resolved by reverse phase-HPLC increased during ripening. No significant differences were found between the sensory properties of cheeses made using a starter, but the cheese made without starter received lower

  18. Carotenoid Profile, Antioxidant Capacity, and Chromoplasts of Pink Guava (Psidium guajava L. Cv. 'Criolla') during Fruit Ripening.

    PubMed

    Rojas-Garbanzo, Carolina; Gleichenhagen, Maike; Heller, Annerose; Esquivel, Patricia; Schulze-Kaysers, Nadine; Schieber, Andreas

    2017-05-10

    Pigments of pericarp and pulp of pink guava (Psidium guajava L. cv. 'Criolla') were investigated to elucidate the profile and the accumulation of main carotenoids during four stages of fruit ripening by using HPLC-DAD and APCI-MS/MS analysis. Seventeen carotenoids were identified, and changes in their profile during fruit ripening were observed. The carotenoids all-trans-β-carotene, 15-cis-lycopene, and all-trans-lycopene were present in all ripening stages, but all-trans-lycopene was found to be predominant (from 63% to 92% of total carotenoids) and responsible for the high lipophilic antioxidant capacity determined by spectrophotometric assays. By using light and transmission electron microscopy, the development of chromoplasts in pericarp and pulp was demonstrated. The accumulation of all-trans-lycopene and all-trans-β-carotene coincided with the development of large crystals; the chromoplasts of pink guava belong, therefore, to the crystalline type.

  19. Fleshy Fruit Expansion and Ripening Are Regulated by the Tomato SHATTERPROOF Gene TAGL1[W][OA

    PubMed Central

    Vrebalov, Julia; Pan, Irvin L.; Arroyo, Antonio Javier Matas; McQuinn, Ryan; Chung, MiYoung; Poole, Mervin; Rose, Jocelyn; Seymour, Graham; Grandillo, Silvana; Giovannoni, James; Irish, Vivian F.

    2009-01-01

    The maturation and ripening of fleshy fruits is a developmental program that synchronizes seed maturation with metabolism, rendering fruit tissues desirable to seed dispersing organisms. Through RNA interference repression, we show that Tomato AGAMOUS-LIKE1 (TAGL1), the tomato (Solanum lycopersicum) ortholog of the duplicated SHATTERPROOF (SHP) MADS box genes of Arabidopsis thaliana, is necessary for fruit ripening. Tomato plants with reduced TAGL1 mRNA produced yellow-orange fruit with reduced carotenoids and thin pericarps. These fruit are also decreased in ethylene, indicating a comprehensive inhibition of maturation mediated through reduced ACC Synthase 2 expression. Furthermore, ectopic expression of TAGL1 in tomato resulted in expansion of sepals and accumulation of lycopene, supporting the role of TAGL1 in ripening. In Arabidopsis, the duplicate SHP1 and SHP2 MADS box genes regulate the development of separation layers essential for pod shatter. Expression of TAGL1 in Arabidopsis failed to completely rescue the shp1 shp2 mutant phenotypes, indicating that TAGL1 has evolved distinct molecular functions compared with its Arabidopsis counterparts. These analyses demonstrate that TAGL1 plays an important role in regulating both fleshy fruit expansion and the ripening process that together are necessary to promote seed dispersal of fleshy fruit. From this broad perspective, SHP1/2 and TAGL1, while distinct in molecular function, regulate similar activities via their necessity for seed dispersal in Arabidopsis and tomato, respectively. PMID:19880793

  20. A Quantitative Chemotherapy Genetic Interaction Map Reveals Factors Associated with PARP Inhibitor Resistance.

    PubMed

    Hu, Hsien-Ming; Zhao, Xin; Kaushik, Swati; Robillard, Lilliane; Barthelet, Antoine; Lin, Kevin K; Shah, Khyati N; Simmons, Andy D; Raponi, Mitch; Harding, Thomas C; Bandyopadhyay, Sourav

    2018-04-17

    Chemotherapy is used to treat most cancer patients, yet our understanding of factors that dictate response and resistance to such drugs remains limited. We report the generation of a quantitative chemical-genetic interaction map in human mammary epithelial cells charting the impact of the knockdown of 625 genes related to cancer and DNA repair on sensitivity to 29 drugs, covering all classes of chemotherapy. This quantitative map is predictive of interactions maintained in other cell lines, identifies DNA-repair factors, predicts cancer cell line responses to therapy, and prioritizes synergistic drug combinations. We identify that ARID1A loss confers resistance to PARP inhibitors in cells and ovarian cancer patients and that loss of GPBP1 causes resistance to cisplatin and PARP inhibitors through the regulation of genes involved in homologous recombination. This map helps navigate patient genomic data and optimize chemotherapeutic regimens by delineating factors involved in the response to specific types of DNA damage. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  1. Persistence of Mycobacterium paratuberculosis during Manufacture and Ripening of Cheddar Cheese

    PubMed Central

    Donaghy, J. A.; Totton, N. L.; Rowe, M. T.

    2004-01-01

    Model Cheddar cheeses were prepared from pasteurized milk artificially contaminated with high 104 to 105 CFU/ml) and low (101 to 102 CFU/ml) inocula of three different Mycobacterium paratuberculosis strains. A reference strain, NCTC 8578, and two strains (806PSS and 796PSS) previously isolated from pasteurized milk for retail sale were investigated in this study. The manufactured Cheddar cheeses were similar in pH, salt, moisture, and fat composition to commercial Cheddar. The survival of M. paratuberculosis cells was monitored over a 27-week ripening period by plating homogenized cheese samples onto HEYM agar medium supplemented with the antibiotics vancomycin, amphotericin B, and nalidixic acid without a decontamination step. A concentration effect was observed in M. paratuberculosis numbers between the inoculated milk and the 1-day old cheeses for each strain. For all manufactured cheeses, a slow gradual decrease in M. paratuberculosis CFU in cheese was observed over the ripening period. In all cases where high levels (>3.6 log10) of M. paratuberculosis were present in 1-day cheeses, the organism was culturable after the 27-week ripening period. The D values calculated for strains 806PSS, 796PSS, and NCTC 8578 were 107, 96, and 90 days, respectively. At low levels of contamination, M. paratuberculosis was only culturable from 27-week-old cheese spiked with strain 806PSS. M. paratuberculosis was recovered from the whey fraction in 10 of the 12 manufactured cheeses. Up to 4% of the initial M. paratuberculosis load was recovered in the culture-positive whey fractions at either the high or low initial inoculum. PMID:15294829

  2. Metabolic Profiling during Peach Fruit Development and Ripening Reveals the Metabolic Networks That Underpin Each Developmental Stage1[C][W

    PubMed Central

    Lombardo, Verónica A.; Osorio, Sonia; Borsani, Julia; Lauxmann, Martin A.; Bustamante, Claudia A.; Budde, Claudio O.; Andreo, Carlos S.; Lara, María V.; Fernie, Alisdair R.; Drincovich, María F.

    2011-01-01

    Fruit from rosaceous species collectively display a great variety of flavors and textures as well as a generally high content of nutritionally beneficial metabolites. However, relatively little analysis of metabolic networks in rosaceous fruit has been reported. Among rosaceous species, peach (Prunus persica) has stone fruits composed of a juicy mesocarp and lignified endocarp. Here, peach mesocarp metabolic networks were studied across development using metabolomics and analysis of key regulatory enzymes. Principal component analysis of peach metabolic composition revealed clear metabolic shifts from early through late development stages and subsequently during postharvest ripening. Early developmental stages were characterized by a substantial decrease in protein abundance and high levels of bioactive polyphenols and amino acids, which are substrates for the phenylpropanoid and lignin pathways during stone hardening. Sucrose levels showed a large increase during development, reflecting translocation from the leaf, while the importance of galactinol and raffinose is also inferred. Our study further suggests that posttranscriptional mechanisms are key for metabolic regulation at early stages. In contrast to early developmental stages, a decrease in amino acid levels is coupled to an induction of transcripts encoding amino acid and organic acid catabolic enzymes during ripening. These data are consistent with the mobilization of amino acids to support respiration. In addition, sucrose cycling, suggested by the parallel increase of transcripts encoding sucrose degradative and synthetic enzymes, appears to operate during postharvest ripening. When taken together, these data highlight singular metabolic programs for peach development and may allow the identification of key factors related to agronomic traits of this important crop species. PMID:22021422

  3. Two divergent endo-beta-1,4-glucanase genes exhibit overlapping expression in ripening fruit and abscising flowers.

    PubMed Central

    Lashbrook, C C; Gonzalez-Bosch, C; Bennett, A B

    1994-01-01

    Two structurally divergent endo-beta-1,4-glucanase (EGase) cDNAs were cloned from tomato. Although both cDNAs (Cel1 and Cel2) encode potentially glycosylated, basic proteins of 51 to 53 kD and possess multiple amino acid domains conserved in both plant and microbial EGases, Cel1 and Cel2 exhibit only 50% amino acid identity at the overall sequence level. Amino acid sequence comparisons to other plant EGases indicate that tomato Cel1 is most similar to bean abscission zone EGase (68%), whereas Cel2 exhibits greatest sequence identity to avocado fruit EGase (57%). Sequence comparisons suggest the presence of at least two structurally divergent EGase families in plants. Unlike ripening avocado fruit and bean abscission zones in which a single EGase mRNA predominates, EGase expression in tomato reflects the overlapping accumulation of both Cel1 and Cel2 transcripts in ripening fruit and in plant organs undergoing cell separation. Cel1 mRNA contributes significantly to total EGase mRNA accumulation within plant organs undergoing cell separation (abscission zones and mature anthers), whereas Cel2 mRNA is most abundant in ripening fruit. The overlapping expression of divergent EGase genes within a single species may suggest that multiple activities are required for the cooperative disassembly of cell wall components during fruit ripening, floral abscission, and anther dehiscence. PMID:7994180

  4. Mining secreted proteins that function in pepper fruit development and ripening using a yeast secretion trap (YST)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lee, Je Min, E-mail: jemin@knu.ac.kr; Department of Horticultural Science, Kyungpook National University, Daegu; Lee, Sang-Jik

    Highlights: • Yeast secretion trap (YST) is a valuable tool for mining secretome. • A total of 80 secreted proteins are newly identified via YST in pepper fruits. • The secreted proteins are differentially regulated during pepper development and ripening. • Transient GFP-fusion assay and in planta secretion trap can effectively validate the secretion of proteins. - Abstract: Plant cells secrete diverse sets of constitutively- and conditionally-expressed proteins under various environmental and developmental states. Secreted protein populations, or secretomes have multiple functions, including defense responses, signaling, metabolic processes, and developmental regulation. To identify genes encoding secreted proteins that function inmore » fruit development and ripening, a yeast secretion trap (YST) screen was employed using pepper (Capsicum annuum) fruit cDNAs. The YST screen revealed 80 pepper fruit-related genes (CaPFRs) encoding secreted proteins including cell wall proteins, several of which have not been previously described. Transient GFP-fusion assay and an in planta secretion trap were used to validate the secretion of proteins encoded by selected YST clones. In addition, RNA gel blot analyses provided further insights into their expression and regulation during fruit development and ripening. Integrating our data, we conclude that the YST provides a valuable functional genomics tool for the identification of substantial numbers of novel secreted plant proteins that are associated with biological processes, including fruit development and ripening.« less

  5. Immunization by bovine thrombin used with fibrin glue during cardiovascular operations. Development of thrombin and factor V inhibitors.

    PubMed

    Berruyer, M; Amiral, J; Ffrench, P; Belleville, J; Bastien, O; Clerc, J; Kassir, A; Estanove, S; Dechavanne, M

    1993-05-01

    Brief case histories of three patients aged 58, 38, and 44 years are reported. All underwent cardiovascular operations. Subsequently hemostasis test abnormalities developed between the seventh and eighth postoperative days after exposure to bovine thrombin used with fibrin glue. These were characterized by an increased activated partial thromboplastin time (64 to 147 seconds), prothrombin time (19 to 24 seconds), bovine thrombin time (> 120 seconds) and a markedly reduced factor V level (< 10% in two patients and 16% in the third patient). A patient plasma dilution of 1 in 200 with a normal plasma pool was necessary to correct bovine thrombin time. No fast-acting or progressive inhibitor against factor V could be detected by coagulation tests, and fresh frozen plasma perfusion had no effect. Plasmapheresis was performed preventatively to avoid bleeding, and factor V levels stabilized at around 50% after two to four exchanges. Immunologic studies showed that the inhibitors were directed not only against bovine factors but also against human ones. Therefore factor V decrease could have been the result of rapid clearance from the circulation of complexes formed with a nonneutralizing inhibitor that is not detected by clotting tests. These antibodies were purified by standard methods and immunoaffinity. Fast immunization could be explained by a prior sensitization to bovine thrombin exposure during previous operations. It is suggested that bovine thrombin used with fibrin glue contains small amounts of factor V and may be responsible for these abnormalities. This is in agreement with previous literature reports. However, these described neutralizing factor V inhibitors, which were easily detected.

  6. Altered Middle Lamella Homogalacturonan and Disrupted Deposition of (1→5)-α-l-Arabinan in the Pericarp of Cnr, a Ripening Mutant of Tomato1

    PubMed Central

    Orfila, Caroline; Seymour, Graham B.; Willats, William G.T.; Huxham, I. Max; Jarvis, Michael C.; Dover, Colin J.; Thompson, Andrew J.; Knox, J. Paul

    2001-01-01

    Cnr (colorless non-ripening) is a pleiotropic tomato (Lycopersicon esculentum) fruit ripening mutant with altered tissue properties including weaker cell-to-cell contacts in the pericarp (A.J. Thompson, M. Tor, C.S. Barry, J. Vrebalov, C. Orfila, M.C. Jarvis, J.J. Giovannoni, D. Grierson, G.B. Seymour [1999] Plant Physiol 120: 383–390). Whereas the genetic basis of the Cnr mutation is being identified by molecular analyses, here we report the identification of cell biological factors underlying the Cnr texture phenotype. In comparison with wild type, ripe-stage Cnr fruits have stronger, non-swollen cell walls (CW) throughout the pericarp and extensive intercellular space in the inner pericarp. Using electron energy loss spectroscopy imaging of calcium-binding capacity and anti-homogalacturonan (HG) antibody probes (PAM1 and JIM5) we demonstrate that maturation processes involving middle lamella HG are altered in Cnr fruit, resulting in the absence or a low level of HG-/calcium-based cell adhesion. We also demonstrate that the deposition of (1→5)-α-l-arabinan is disrupted in Cnr pericarp CW and that this disruption occurs prior to fruit ripening. The relationship between the disruption of (1→5)-α-l-arabinan deposition in pericarp CW and the Cnr phenotype is discussed. PMID:11351084

  7. Mango (Mangifera indica L.) cv. Kent fruit mesocarp de novo transcriptome assembly identifies gene families important for ripening

    PubMed Central

    Dautt-Castro, Mitzuko; Ochoa-Leyva, Adrian; Contreras-Vergara, Carmen A.; Pacheco-Sanchez, Magda A.; Casas-Flores, Sergio; Sanchez-Flores, Alejandro; Kuhn, David N.; Islas-Osuna, Maria A.

    2015-01-01

    Fruit ripening is a physiological and biochemical process genetically programmed to regulate fruit quality parameters like firmness, flavor, odor and color, as well as production of ethylene in climacteric fruit. In this study, a transcriptomic analysis of mango (Mangifera indica L.) mesocarp cv. “Kent” was done to identify key genes associated with fruit ripening. Using the Illumina sequencing platform, 67,682,269 clean reads were obtained and a transcriptome of 4.8 Gb. A total of 33,142 coding sequences were predicted and after functional annotation, 25,154 protein sequences were assigned with a product according to Swiss-Prot database and 32,560 according to non-redundant database. Differential expression analysis identified 2,306 genes with significant differences in expression between mature-green and ripe mango [1,178 up-regulated and 1,128 down-regulated (FDR ≤ 0.05)]. The expression of 10 genes evaluated by both qRT-PCR and RNA-seq data was highly correlated (R = 0.97), validating the differential expression data from RNA-seq alone. Gene Ontology enrichment analysis, showed significantly represented terms associated to fruit ripening like “cell wall,” “carbohydrate catabolic process” and “starch and sucrose metabolic process” among others. Mango genes were assigned to 327 metabolic pathways according to Kyoto Encyclopedia of Genes and Genomes database, among them those involved in fruit ripening such as plant hormone signal transduction, starch and sucrose metabolism, galactose metabolism, terpenoid backbone, and carotenoid biosynthesis. This study provides a mango transcriptome that will be very helpful to identify genes for expression studies in early and late flowering mangos during fruit ripening. PMID:25741352

  8. Phenolic Profiles, Antioxidant Activities, and Neuroprotective Properties of Mulberry (Morus atropurpurea Roxb.) Fruit Extracts from Different Ripening Stages.

    PubMed

    Yang, Jiufang; Liu, Xuanjun; Zhang, Xiaoxu; Jin, Qing; Li, Jingming

    2016-10-01

    The present work investigated the phenolic profiles (including nonanthocyanin and anthocyanin phenolics), antioxidant activities, and neuroprotective potential of mulberry fruit (MF) (Morus atropurpurea Roxb.) grown in China at different ripening stages. High-performance liquid chromatography-tandem mass spectrometry method (HPLC-MS/MS) was used to identify and quantify the phenolic compounds. The antioxidant capacity, total phenolic content (TPC), total flavonoid content (TFC), and total monomeric anthocyanin content (TAC) were determined using spectrophotometric methods. The neuroprotective effects of MFs at different ripening stages were investigated using Aβ 25-35 -treated PC12 cells as the cellular model of Alzheimer's disease. Of the 19 phenolic compounds characterized from the MF extracts, the contents of rutin and anthocyanins increased and that of chlorogenic acid decreased significantly with maturity. At the fully ripened stage, MF extracts showed the highest amounts of TPC (11.23 mg gallic acid equivalents/g fresh weight), TFC (15.1 mg rutin equivalents/g fresh weight), and TAC (1177 mg cyanidin 3-O-glucoside equivalents/100 g fresh weight). Meanwhile, antioxidant activity of MF extracts at this stage was highest according to ABTS (an IC50 value of 4.11 μg/mL) and DPPH (an IC50 value of 10.08 μg/mL) assays. Cellular assays revealed increased cell viability in cells treated with the ripe MF extracts; compared with the control groups, the ripening fruits also increased the antioxidant enzyme levels in PC12 cells. Together, these results suggest that the antioxidant activities and neuroprotective properties of ripening MFs are related to the contents and types of phenolic compounds that are present in the fruits. © 2016 Institute of Food Technologists®.

  9. Products Released from Enzymically Active Cell Wall Stimulate Ethylene Production and Ripening in Preclimacteric Tomato (Lycopersicon esculentum Mill.) Fruit 1

    PubMed Central

    Brecht, Jeffrey K.; Huber, Donald J.

    1988-01-01

    Enzymically active cell wall from ripe tomato (Lycopersicon esculentum Mill.) fruit pericarp release uronic acids through the action of wall-bound polygalacturonase. The potential involvement of products of wall hydrolysis in the induction of ethylene synthesis during tomato ripening was investigated by vacuum infiltrating preclimacteric (green) fruit with solutions containing pectin fragments enzymically released from cell wall from ripe fruit. Ripening initiation was accelerated in pectin-infiltrated fruit compared to control (buffer-infiltrated) fruit as measured by initiation of climacteric CO2 and ethylene production and appearance of red color. The response to infiltration was maximum at a concentration of 25 micrograms pectin per fruit; higher concentrations (up to 125 micrograms per fruit) had no additional effect. When products released from isolated cell wall from ripe pericarp were separated on Bio-Gel P-2 and specific size classes infiltrated into preclimacteric fruit, ripening-promotive activity was found only in the larger (degree of polymerization >8) fragments. Products released from pectin derived from preclimacteric pericarp upon treatment with polygalacturonase from ripe pericarp did not stimulate ripening when infiltrated into preclimacteric fruit. PMID:16666417

  10. In Depth Proteome Analysis of Ripening Muscadine Grape Berry cv. Carlos Reveals Proteins Associated with Flavor and Aroma Compounds.

    PubMed

    Kambiranda, Devaiah; Basha, Sheikh M; Singh, Rakesh K; He, Huan; Calvin, Kate; Mercer, Roger

    2016-09-02

    Ripening in nonclimacteric fruits such as grape involves complex chemical changes that have a profound influence on the accumulation of flavor and aroma compounds distinct to a particular grape genotype. In this study, proteome characterization of wine type bronze muscadine grape (Vitis rotundifolia cv. Carlos), primarily grown in the Southeastern United States was performed during berry ripening. Stage-specific protein expression was obtained among different stages of berries. Differential analysis showed the expression of 522 proteins that regulate diverse biological processes and metabolic pathways. Of these, 30 proteins are associated with the production of key phenolic compounds, whereas 25 are associated with the production of muscadine aroma compounds. These proteins are involved in the phenylpropanoid pathway, terpene synthesis, fatty acid derived volatiles and esters that affect muscadine berry flavor and aroma characteristics. Further, gene expression analysis during ripening validated the expression pattern of 12 proteins. Catechin, epicatechin, and four stilbenes were quantified to correlate observed proteome changes. This study not only revealed biochemical changes during muscadine berry ripening but also offers indicators for marker-assisted breeding to enhance organoleptic properties of muscadine grape to improve its flavor and aroma properties.

  11. Hormones, polyamines, and cell wall metabolism during oil palm fruit mesocarp development and ripening.

    PubMed

    Teh, Huey Fang; Neoh, Bee Keat; Wong, Yick Ching; Kwong, Qi Bin; Ooi, Tony Eng Keong; Ng, Theresa Lee Mei; Tiong, Soon Huat; Low, Jaime Yoke Sum; Danial, Asma Dazni; Ersad, Mohd Amiron; Kulaveerasingam, Harikrishna; Appleton, David R

    2014-08-13

    Oil palm is one of the most productive oil-producing crops and can store up to 90% oil in its fruit mesocarp. Oil palm fruit is a sessile drupe consisting of a fleshy mesocarp from which palm oil is extracted. Biochemical changes in the mesocarp cell walls, polyamines, and hormones at different ripening stages of oil palm fruits were studied, and the relationship between the structural and the biochemical metabolism of oil palm fruits during ripening is discussed. Time-course analysis of the changes in expression of polyamines, hormones, and cell-wall-related genes and metabolites provided insights into the complex processes and interactions involved in fruit development. Overall, a strong reduction in auxin-responsive gene expression was observed from 18 to 22 weeks after pollination. High polyamine concentrations coincided with fruit enlargement during lipid accumulation and latter stages of maturation. The trend of abscisic acid (ABA) concentration was concordant with GA₄ but opposite to the GA₃ profile such that as ABA levels increase the resulting elevated ABA/GA₃ ratio clearly coincides with maturation. Polygalacturonase, expansin, and actin gene expressions were also observed to increase during fruit maturation. The identification of the master regulators of these coordinated processes may allow screening for oil palm variants with altered ripening profiles.

  12. Effect of hypoxia on tissue factor pathway inhibitor expression in breast cancer.

    PubMed

    Cui, X Y; Tinholt, M; Stavik, B; Dahm, A E A; Kanse, S; Jin, Y; Seidl, S; Sahlberg, K K; Iversen, N; Skretting, G; Sandset, P M

    2016-02-01

    ESSENTIALS: A hypoxic microenvironment is a common feature of tumors that may influence activation of coagulation. MCF-7 and SK-BR-3 breast cancer cells and breast cancer tissue samples were used. The results showed transcriptional repression of tissue factor pathway inhibitor expression in hypoxia. Hypoxia-inducible factor 1α may be a target for the therapy of cancer-related coagulation and thrombosis. Activation of coagulation is a common finding in patients with cancer, and is associated with an increased risk of venous thrombosis. As a hypoxic microenvironment is a common feature of solid tumors, we investigated the role of hypoxia in the regulation of tissue factor (TF) pathway inhibitor (TFPI) expression in breast cancer. To explore the transcriptional regulation of TFPI by hypoxia-inducible factor (HIF)-1α in breast cancer cells and their correlation in breast cancer tissues. MCF-7 and SK-BR-3 breast cancer cells were cultured in 1% oxygen or treated with cobalt chloride (CoCl2 ) to mimic hypoxia. Time-dependent and dose-dependent downregulation of TFPI mRNA (quantitative RT-PCR) and of free TFPI protein (ELISA) were observed in hypoxia. Western blotting showed parallel increases in the levels of HIF-1α protein and TF. HIF-1α inhibitor abolished or attenuated the hypoxia-induced downregulation of TFPI. Luciferase reporter assay showed that both hypoxia and HIF-1α overexpression caused strong repression of TFPI promoter activity. Subsequent chromatin immunoprecipitation and mutagenesis analysis demonstrated a functional hypoxia response element within the TFPI promoter, located at -1065 to -1060 relative to the transcriptional start point. In breast cancer tissue samples, gene expression analyses showed a positive correlation between the mRNA expression of TFPI and that of HIF-1α. This study demonstrates that HIF-1α is involved in the transcriptional regulation of the TFPI gene, and suggests that a hypoxic microenvironment inside a breast tumor may

  13. Novel factor Xa inhibitors: a patent review.

    PubMed

    de Candia, Modesto; Lopopolo, Gianfranco; Altomare, Cosimo

    2009-11-01

    New oral anticoagulants with favorable safety profiles and fixed doses are required for the management of thromboembolism and stroke prevention in patients with atrial fibrillation. Among them, fXa inhibitors (the so-called xabans) are attractive options that can overcome limitations (e.g., bleeding) of the current oral antithrombotic therapy. The rational design of small-molecule direct fXa inhibitors, whose importance is testified by the growing number of publications and patents recently registered, has been fully supported by the X-ray crystallography of enzyme-ligand complexes. Pubmed, SciFinder Scholar, ISI web of knowledge(SM), http://ep.espacenet.com/ and Google websites were used as the main sources for literature retrieving, and > 100 patents filed between 2006 and April 2009, reviewed and discussed herein, highlight the variety among the P1 and P4 moieties on suitable scaffolds. The replacement of the benzamidine P1 moiety, which characterizes the first generation, with less basic bioisosteric or nonpolar neutral P1 groups led to the disclosure of numerous fXa inhibitors with high potency, selectivity and oral bioavailability. Novel selective fXa inhibitors with stable pharmacokinetics, better therapeutic windows and ease-of-use than the existing anticoagulants are currently under advanced stage clinical trials. Available data from Phase II and Phase III studies reflect the drive towards fXa inhibitors as potentially more effective and safer antithrombotic drugs. Their development is expected to address two major needs for anticoagulation, namely safety and ease-of-use, and to significantly affect the anticoagulant market.

  14. Synergistic inhibition with a dual epidermal growth factor receptor/HER-2/neu tyrosine kinase inhibitor and a disintegrin and metalloprotease inhibitor.

    PubMed

    Witters, Lois; Scherle, Peggy; Friedman, Steven; Fridman, Jordan; Caulder, Eian; Newton, Robert; Lipton, Allan

    2008-09-01

    The ErbB family of receptors is overexpressed in numerous human tumors. Overexpression correlates with poor prognosis and resistance to therapy. Use of ErbB-specific antibodies to the receptors (Herceptin or Erbitux) or ErbB-specific small-molecule inhibitors of the receptor tyrosine kinase activity (Iressa or Tarceva) has shown clinical efficacy in several solid tumors. An alternative method of affecting ErbB-initiated tumor growth and survival is to block sheddase activity. Sheddase activity is responsible for cleavage of multiple ErbB ligands and receptors, a necessary step in availability of the soluble, active form of the ligand and a constitutively activated ligand-independent receptor. This sheddase activity is attributed to the ADAM (a disintegrin and metalloprotease) family of proteins. ADAM 10 is the main sheddase of epidermal growth factor (EGF) and HER-2/neu cleavage, whereas ADAM17 is required for cleavage of additional EGF receptor (EGFR) ligands (transforming growth factor-alpha, amphiregulin, heregulin, heparin binding EGF-like ligand). This study has shown that addition of INCB3619, a potent inhibitor of ADAM10 and ADAM17, reduces in vitro HER-2/neu and amphiregulin shedding, confirming that it interferes with both HER-2/neu and EGFR ligand cleavage. Combining INCB3619 with a lapatinib-like dual inhibitor of EGFR and HER-2/neu kinases resulted in synergistic growth inhibition in MCF-7 and HER-2/neu-transfected MCF-7 human breast cancer cells. Combining the INCB7839 second-generation sheddase inhibitor with lapatinib prevented the growth of HER-2/neu-positive BT474-SC1 human breast cancer xenografts in vivo. These results suggest that there may be an additional clinical benefit of combining agents that target the ErbB pathways at multiple points.

  15. Genome-wide analysis and characterization of Aux/IAA family genes related to fruit ripening in papaya (Carica papaya L.).

    PubMed

    Liu, Kaidong; Yuan, Changchun; Feng, Shaoxian; Zhong, Shuting; Li, Haili; Zhong, Jundi; Shen, Chenjia; Liu, Jinxiang

    2017-05-05

    Auxin/indole-3-acetic acid (Aux/IAA) family genes encode short-lived nuclear proteins that mediate the responses of auxin-related genes and are involved in several plant developmental and growth processes. However, how Aux/IAA genes function in the fruit development and ripening of papaya (Carica papaya L.) is largely unknown. In this study, a comprehensive identification and a distinctive expression analysis of 18 C. papaya Aux/IAA (CpIAA) genes were performed using newly updated papaya reference genome data. The Aux/IAA gene family in papaya is slightly smaller than that in Arabidopsis, but all of the phylogenetic subfamilies are represented. Most of the CpIAA genes are responsive to various phytohormones and expressed in a tissues-specific manner. To understand the putative biological functions of the CpIAA genes involved in fruit development and ripening, quantitative real-time PCR was used to test the expression profiling of CpIAA genes at different stages. Furthermore, an IAA treatment significantly delayed the ripening process in papaya fruit at the early stages. The expression changes of CpIAA genes in ACC and 1-MCP treatments suggested a crosstalk between auxin and ethylene during the fruit ripening process of papaya. Our study provided comprehensive information on the Aux/IAA family in papaya, including gene structures, phylogenetic relationships and expression profiles. The involvement of CpIAA gene expression changes in fruit development and ripening gives us an opportunity to understand the roles of auxin signaling in the maturation of papaya reproductive organs.

  16. Acrylamide formation in plantain (Musa paradisiaca) chips influenced by different ripening stages: A correlation study with respect to reducing sugars, amino acids and phenolic content.

    PubMed

    Shamla, L; Nisha, P

    2017-05-01

    The effect of ripening on the formation of acrylamide in deep fried plantain chips made from Nendran variety (Musa paradisiaca) was investigated. The precursors of acrylamide formation, reducing sugars (glucose and fructose) and ten major amino acids, were quantified during different stages of ripening using HPLC and correlated with acrylamide formation. The total phenolic content and total flavonoid content were also estimated and correlated with acrylamide formation. Both glucose and fructose increased during ripening and demonstrated a positive correlation on formation of acrylamide (correlation coefficient of r=0.95 and 0.94 respectively (p<0.05), whereas asparagine, was poorly correlated (p>0.05). The decreased levels of phenolic content during ripening of plantain were negatively correlated with acrylamide formation in the deep fried chips prepared. Thus the selection of proper ripening stage renders reduced formation of acrylamide in plantain chips to a reasonable extend. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. A specific antidote for reversal of anticoagulation by direct and indirect inhibitors of coagulation factor Xa.

    PubMed

    Lu, Genmin; DeGuzman, Francis R; Hollenbach, Stanley J; Karbarz, Mark J; Abe, Keith; Lee, Gail; Luan, Peng; Hutchaleelaha, Athiwat; Inagaki, Mayuko; Conley, Pamela B; Phillips, David R; Sinha, Uma

    2013-04-01

    Inhibitors of coagulation factor Xa (fXa) have emerged as a new class of antithrombotics but lack effective antidotes for patients experiencing serious bleeding. We designed and expressed a modified form of fXa as an antidote for fXa inhibitors. This recombinant protein (r-Antidote, PRT064445) is catalytically inactive and lacks the membrane-binding γ-carboxyglutamic acid domain of native fXa but retains the ability of native fXa to bind direct fXa inhibitors as well as low molecular weight heparin-activated antithrombin III (ATIII). r-Antidote dose-dependently reversed the inhibition of fXa by direct fXa inhibitors and corrected the prolongation of ex vivo clotting times by such inhibitors. In rabbits treated with the direct fXa inhibitor rivaroxaban, r-Antidote restored hemostasis in a liver laceration model. The effect of r-Antidote was mediated by reducing plasma anti-fXa activity and the non-protein bound fraction of the fXa inhibitor in plasma. In rats, r-Antidote administration dose-dependently and completely corrected increases in blood loss resulting from ATIII-dependent anticoagulation by enoxaparin or fondaparinux. r-Antidote has the potential to be used as a universal antidote for a broad range of fXa inhibitors.

  18. A Ser/Thr protein kinase phosphorylates MA-ACS1 (Musa acuminata 1-aminocyclopropane-1-carboxylic acid synthase 1) during banana fruit ripening.

    PubMed

    Choudhury, Swarup Roy; Roy, Sujit; Sengupta, Dibyendu N

    2012-08-01

    1-Aminocyclopropane-1-carboxylic acid synthase (ACS) catalyzes the rate-limiting step in ethylene biosynthesis during ripening. ACS isozymes are regulated both transcriptionally and post-translationally. However, in banana, an important climacteric fruit, little is known about post-translational regulation of ACS. Here, we report the post-translational modification of MA-ACS1 (Musa acuminata ACS1), a ripening inducible isozyme in the ACS family, which plays a key role in ethylene biosynthesis during banana fruit ripening. Immunoprecipitation analyses of phospholabeled protein extracts from banana fruit using affinity-purified anti-MA-ACS1 antibody have revealed phosphorylation of MA-ACS1, particularly in ripe fruit tissue. We have identified the induction of a 41-kDa protein kinase activity in pulp at the onset of ripening. The 41-kDa protein kinase has been identified as a putative protein kinase by MALDI-TOF/MS analysis. Biochemical analyses using partially purified protein kinase fraction from banana fruit have identified the protein kinase as a Ser/Thr family of protein kinase and its possible involvement in MA-ACS1 phosphorylation during ripening. In vitro phosphorylation analyses using synthetic peptides and site-directed mutagenized recombinant MA-ACS1 have revealed that serine 476 and 479 residues at the C-terminal region of MA-ACS1 are phosphorylated. Overall, this study provides important novel evidence for in vivo phosphorylation of MA-ACS1 at the molecular level as a possible mechanism of post-translational regulation of this key regulatory protein in ethylene signaling pathway in banana fruit during ripening.

  19. EIN3-like gene expression during fruit ripening of Cavendish banana (Musa acuminata cv. Grande naine).

    PubMed

    Mbéguié-A-Mbéguié, Didier; Hubert, Olivier; Fils-Lycaon, Bernard; Chillet, Marc; Baurens, Franc-Christophe

    2008-06-01

    Ethylene signal transduction initiates with ethylene binding at receptor proteins and terminates in a transcription cascade involving the EIN3/EIL transcription factors. Here, we have isolated four cDNAs homologs of the Arabidopsis EIN3/EIN3-like gene, MA-EILs (Musa acuminata ethylene insensitive 3-like) from banana fruit. Sequence comparison with other banana EIL gene already registered in the database led us to conclude that, at this day, at least five different genes namely MA-EIL1, MA-EIL2/AB266318, MA-EIL3/AB266319, MA-EIL4/AB266320 and AB266321 exist in banana. Phylogenetic analyses included all banana EIL genes within a same cluster consisting of rice OsEILs, a monocotyledonous plant as banana. However, MA-EIL1, MA-EIL2/AB266318, MA-EIL4/AB266320 and AB266321 on one side, and MA-EIL3/AB266319 on the other side, belong to two distant subclusters. MA-EIL mRNAs were detected in all examined banana tissues but at lower level in peel than in pulp. According to tissues, MA-EIL genes were differentially regulated by ripening and ethylene in mature green fruit and wounding in old and young leaves. MA-EIL2/AB266318 was the unique ripening- and ethylene-induced gene; MA-EIL1, MA-EIL4/Ab266320 and AB266321 genes were downregulated, while MA-EIL3/AB266319 presented an unusual pattern of expression. Interestingly, a marked change was observed mainly in MA-EIL1 and MA-EIL3/Ab266319 mRNA accumulation concomitantly with changes in ethylene responsiveness of fruit. Upon wounding, the main effect was observed in MA-EIL4/AB266320 and AB266321 mRNA levels, which presented a markedly increase in both young and old leaves, respectively. Data presented in this study suggest the importance of a transcriptionally step control in the regulation of EIL genes during banana fruit ripening.

  20. Recombinant factor VIIa (eptacog alfa): a review of its use in congenital hemophilia with inhibitors, acquired hemophilia, and other congenital bleeding disorders.

    PubMed

    Croom, Katherine F; McCormack, Paul L

    2008-01-01

    Recombinant factor VIIa (NovoSeven; also known as recombinant activated factor VII or eptacog alfa) is structurally similar to human plasma-derived coagulation factor VIIa, but is manufactured using DNA biotechnology. Recombinant factor VIIa interacts with thrombin-activated platelets to produce a thrombin burst leading to accelerated fibrin clot formation localized to the site of vascular injury. It is approved in many countries for use as an intravenous hemostatic agent in patients with congenital hemophilia with inhibitors, and also for acquired hemophilia, factor VII deficiency, and Glanzmann thrombasthenia in some countries. Studies have shown it to be effective and generally well tolerated when used intravenously to treat bleeding episodes or provide hemostatic cover during surgery in patients with congenital hemophilia with inhibitors, acquired hemophilia, factor VII deficiency or Glanzmann thrombasthenia. Based on available data, its efficacy in terms of patient-assessed response may be similar to that of activated prothrombin complex concentrate (aPCC), but treatment with a single 270 microg/kg dose of recombinant factor VIIa might reduce the need for rescue therapy compared with aPCC. Recombinant factor VIIa is not immunogenic in patients with hemophilia, does not produce an anamnestic response in hemophilia patients with inhibitors, and has very low thrombogenicity. It is recommended in guidelines as the treatment of choice for bleeds in patients with hemophilia B with high-responding inhibitors and for patients with factor VII deficiency, and is also a first-line therapeutic option for high-responder hemophilia A patients with inhibitors and those with acquired hemophilia. Cost data from pharmacoeconomic analyses support its use in hemophilia patients with inhibitors. Thus, recombinant factor VIIa is a valuable treatment option for patients with these rare, but potentially serious, bleeding disorders.

  1. Application of infrared microspectroscopy and multivariate analysis for monitoring the effect of adjunct cultures during Swiss cheese ripening.

    PubMed

    Chen, G; Kocaoglu-Vurma, N A; Harper, W J; Rodriguez-Saona, L E

    2009-08-01

    Improved cheese flavor has been attributed to the addition of adjunct cultures, which provide certain key enzymes for proteolysis and affect the dynamics of starter and nonstarter cultures. Infrared microspectroscopy provides unique fingerprint-like spectra for cheese samples and allows for rapid monitoring of cheese composition during ripening. The objective was to use infrared microspectroscopy and multivariate analysis to evaluate the effect of adjunct cultures on Swiss cheeses during ripening. Swiss cheeses, manufactured using a commercial starter culture combination and 1 of 3 adjunct Lactobacillus spp., were evaluated at d 1, 6, 30, 60, and 90 of ripening. Cheese samples (approximately 20 g) were powdered with liquid nitrogen and homogenized using water and organic solvents, and the water-soluble components were separated. A 3-microL aliquot of the extract was applied onto a reflective microscope slide, vacuum-dried, and analyzed by infrared microspectroscopy. The infrared spectra (900 to 1,800 cm(-1)) produced specific absorption profiles that allowed for discrimination among different cheese samples. Cheeses manufactured with adjunct cultures showed more uniform and consistent spectral profiles, leading to the formation of tight clusters by pattern-recognition analysis (soft independent modeling of class analogy) as compared with cheeses with no adjuncts, which exhibited more spectral variability among replicated samples. In addition, the soft independent modeling of class analogy discriminating power indicated that cheeses were differentiated predominantly based on the band at 1,122 cm(-1), which was associated with S-O vibrations. The greatest changes in the chemical profile of each cheese occurred between d 6 and 30 of warm-room ripening. The band at 1,412 cm(-1), which was associated with acidic AA, had the greatest contribution to differentiation, indicating substantial changes in levels of proteolysis during warm-room ripening in addition to propionic

  2. Melasma treatment: A novel approach using a topical agent that contains an anti-estrogen and a vascular endothelial growth factor inhibitor.

    PubMed

    Cohen, Philip R

    2017-04-01

    Melasma is an acquired disorder of pigmentation that presents with asymptomatic symmetric darkening of the face. The pathogenesis of this condition is multifactorial and influenced by several factors including female sex hormones, genetic predisposition and ultraviolet light exposure. The management of melasma is usually directed at more than one of the causative etiologic factors and often incorporates a combination of topical agents, with or without the addition of physical modalities. Estrogen and angiogenesis are significant factors in the etiology of melasma. A useful addition to the therapeutic armentarium for treating melasma would include a topical agent that could effect both of these causative factors. Specifically, a topical preparation consisting of an anti-estrogen and a vascular endothelial growth factor inhibitor would accomplish this goal. Suitable candidates that target estrogen receptors and vascular endothelial growth factor are currently used in medical oncology as systemic antineoplastic agents. The anti-estrogen could be either a selective estrogen receptor modulator (such as tamoxifen or raloxifene) or an aromatase inhibitor (such as anastrozole or letrozole or exemestane). The vascular endothelial growth factor inhibitor would be bevacizumab. In conclusion, a novel-topically administered-therapy for melasma would combine an anti-estrogen and a vascular endothelial growth factor inhibitor. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Sequestration of auxin by the indole-3-acetic acid-amido synthetase GH3-1 in grape berry (Vitis vinifera L.) and the proposed role of auxin conjugation during ripening.

    PubMed

    Böttcher, Christine; Keyzers, Robert A; Boss, Paul K; Davies, Christopher

    2010-08-01

    In fleshy fruit, levels of indole-3-acetic acid (IAA), the most abundant auxin, decline towards the onset of ripening. The application of auxins to immature fruit can delay the ripening processes. However, the mechanisms by which the decrease in endogenous IAA concentrations and the maintenance of low auxin levels in maturing fruit are achieved remain elusive. The transcript of a GH3 gene (GH3-1), encoding for an IAA-amido synthetase which conjugates IAA to amino acids, was detected in grape berries (Vitis vinifera L.). GH3-1 expression increased at the onset of ripening (veraison), suggesting that it might be involved in the establishment and maintenance of low IAA concentrations in ripening berries. Furthermore, this grapevine GH3 gene, responded positively to the combined application of abscisic acid and sucrose and to ethylene, linking it to the control of ripening processes. Levels of IAA-aspartic acid (IAA-Asp), an in vitro product of recombinant GH3-1, rose after veraison and remained high during the following weeks of the ripening phase when levels of free IAA were low. A similar pattern of changes in free IAA and IAA-Asp levels was detected in developing tomatoes (Solanum lycopersicum Mill.), where low concentrations of IAA and an increase in IAA-Asp concentrations coincided with the onset of ripening in this climacteric fruit. Since IAA-Asp might be involved in IAA degradation, the GH3 catalysed formation of this conjugate at, and after, the onset of ripening could represent a common IAA inactivation mechanism in climacteric and non-climacteric fruit which enables ripening.

  4. Changes in transcription of cytokinin metabolism and signalling genes in grape (Vitis vinifera L.) berries are associated with the ripening-related increase in isopentenyladenine.

    PubMed

    Böttcher, Christine; Burbidge, Crista A; Boss, Paul K; Davies, Christopher

    2015-09-16

    Cytokinins are known to play an important role in fruit set and early fruit growth, but their involvement in later stages of fruit development is less well understood. Recent reports of greatly increased cytokinin concentrations in the flesh of ripening kiwifruit (Actinidia deliciosa (A. Chev.) C.F. Liang & A.R. Ferguson) and grapes (Vitis vinifera L.) have suggested that these hormones are implicated in the control of ripening-related processes. A similar pattern of isopentenyladenine (iP) accumulation was observed in the ripening fruit of several grapevine cultivars, strawberry (Fragaria ananassa Duch.) and tomato (Solanum lycopersicum Mill.), suggesting a common, ripening-related role for this cytokinin. Significant differences in maximal iP concentrations between grapevine cultivars and between fruit species might reflect varying degrees of relevance or functional adaptations of this hormone in the ripening process. Grapevine orthologues of five Arabidopsis (Arabidopsis thaliana L.) gene families involved in cytokinin metabolism and signalling were identified and analysed for their expression in developing grape berries and a range of other grapevine tissues. Members of each gene family were characterised by distinct expression profiles during berry development and in different grapevine organs, suggesting a complex regulation of cellular cytokinin activities throughout the plant. The post-veraison-specific expression of a set of biosynthesis, activation, perception and signalling genes together with a lack of expression of degradation-related genes during the ripening phase were indicative of a local control of berry iP concentrations leading to the observed accumulation of iP in ripening grapes. The transcriptional analysis of grapevine genes involved in cytokinin production, degradation and response has provided a possible explanation for the ripening-associated accumulation of iP in grapes and other fruit. The pre- and post-veraison-specific expression of

  5. Energetic factors determining the binding of type I inhibitors to c-Met kinase: experimental studies and quantum mechanical calculations.

    PubMed

    Yu, Zhe; Ma, Yu-chi; Ai, Jing; Chen, Dan-qi; Zhao, Dong-mei; Wang, Xin; Chen, Yue-lei; Geng, Mei-yu; Xiong, Bing; Cheng, Mao-sheng; Shen, Jing-Kang

    2013-11-01

    To decipher the molecular interactions between c-Met and its type I inhibitors and to facilitate the design of novel c-Met inhibitors. Based on the prototype model inhibitor 1, four ligands with subtle differences in the fused aromatic rings were synthesized. Quantum chemistry was employed to calculate the binding free energy for each ligand. Symmetry-adapted perturbation theory (SAPT) was used to decompose the binding energy into several fundamental forces to elucidate the determinant factors. Binding free energies calculated from quantum chemistry were correlated well with experimental data. SAPT calculations showed that the predominant driving force for binding was derived from a sandwich π-π interaction with Tyr-1230. Arg-1208 was the differentiating factor, interacting with the 6-position of the fused aromatic ring system through the backbone carbonyl with a force pattern similar to hydrogen bonding. Therefore, a hydrogen atom must be attached at the 6-position, and changing the carbon atom to nitrogen caused unfavorable electrostatic interactions. The theoretical studies have elucidated the determinant factors involved in the binding of type I inhibitors to c-Met.

  6. Gene expression and enzymatic activity of pectin methylesterase during fruit development and ripening in Coffea arabica L.

    PubMed

    Cação, S M B; Leite, T F; Budzinski, I G F; dos Santos, T B; Scholz, M B S; Carpentieri-Pipolo, V; Domingues, D S; Vieira, L G E; Pereira, L F P

    2012-09-03

    Coffee quality is directly related to the harvest and post harvest conditions. Non-uniform maturation of coffee fruits, combined with inadequate harvest, negatively affects the final quality of the product. Pectin methylesterase (PME) plays an important role in fruit softening due to the hydrolysis of methylester groups in cell wall pectins. In order to characterize the changes occurring during coffee fruit maturation, the enzymatic activity of PME was measured during different stages of fruit ripening. PME activity progressively increased from the beginning of the ripening process to the cherry fruit stage. In silico analysis of expressed sequence tags of the Brazilian Coffee Genome Project database identified 5 isoforms of PME. We isolated and cloned a cDNA homolog of PME for further characterization. CaPME4 transcription was analyzed in pericarp, perisperm, and endosperm tissues during fruit development and ripening as well as in other plant tissues. Northern blot analysis revealed increased transcription of CaPME4 in the pericarp 300 days after flowering. Low levels of CaPME4 mRNAs were observed in the endosperm 270 days after flowering. Expression of CaPME4 transcripts was strong in the branches and lower in root and flower tissues. We showed that CaPME4 acts specifically during the later stages of fruit ripening and possibly contributes to the softening of coffee fruit, thus playing a significant role in pectin degradation in the fruit pericarp.

  7. [Comparison of dinoprostone (ovules and gel) to achieve cervical ripening in patients with term pregnancy that occurs with premature membranes rupture].

    PubMed

    López-Farfán, José Angel; Gámez-Guevara, Catalina

    2010-02-01

    Premature rupture of membranes is a normal occurrence of labor and can occur before or after the onset of contractions. The clinical factors associated with premature rupture of membranes include: low socioeconomic status, low body mass index, prior preterm pregnancies, smoking, sexually transmitted infections and urinary tract, conization, cervical cerclage and amniocentesis. To evaluate whether prolonged release of the vaginal insert of PGE2 is superior to dinoprostone gel to achieve cervical ripening in patients with term pregnancy that occur with premature rupture of membranes. Randomized clinical trial in the surgical unit of play in a period of 6 months, with an estimated sample of 50 patients was randomized by block table. After assessment confirming rupture of membranes, Bishop Score and meeting inclusion criteria, group A was applied PGE2 intracervical gel 0.5 mg with a maximum of 3 doses, every 6 hours. Group B was administered at vaginal insert of PGE2 single dose for 24 hours, the patient was left to sleep 30 minutes cardio toco-monitoring chart for at least 2 hours after application. The average time to maturity was 310.59 minutes with a standard deviation of 198.7 and concluded that there was no significant difference between the onset of uterine activity and the onset of labor among the prolonged release dinoprostone and alternatives such as the gel cervical for cervical ripening. Either this is a good choice to ripen the cervix in patients with term pregnancy and premature rupture of membranes.

  8. Utilization of a freeze-thaw treatment to enhance phenolic ripening and tannin oxidation of grape seeds in red (Vitis vinifera L.) cultivars.

    PubMed

    Rustioni, Laura; Cola, Gabriele; VanderWeide, Josh; Murad, Patrick; Failla, Osvaldo; Sabbatini, Paolo

    2018-09-01

    Phenolic ripening represents a major interest for quality wine producers. Nevertheless, climatic or genotypical limitations can often prevent optimal maturation process. During winemaking seeds can be easily separated and technologically processed to improve their quality. Relying on the key role of oxidation for phenolic ripening, a freeze-thaw treatment was proposed to improve the fruit quality for potential use in challenging growing conditions. The experiment was carried on in two distinctive viticultural areas, Michigan and Italy. Five cultivars (Cabernet Franc, Cabernet Sauvignon, Merlot, Pinot noir and Chambourcin) and six cultivars (Cabernet Sauvignon, Sangiovese, Syrah, Croatina, Barbera and Nebbiolo) were used in Michigan and Italy, respectively. Samples were collected at different phenological stages, to describe the natural ripening process and grape seeds were characterized before and after a freeze-thaw treatment. Colorimetric and spectrophotometric data highlighted similarities among natural and artificial seed ripening promising future applications for the wine industries. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. The composition of Camembert cheese-ripening cultures modulates both mycelial growth and appearance.

    PubMed

    Lessard, Marie-Hélène; Bélanger, Gaétan; St-Gelais, Daniel; Labrie, Steve

    2012-03-01

    The fungal microbiota of bloomy-rind cheeses, such as Camembert, forms a complex ecosystem that has not been well studied, and its monitoring during the ripening period remains a challenge. One limitation of enumerating yeasts and molds on traditional agar media is that hyphae are multicellular structures, and colonies on a petri dish rarely develop from single cells. In addition, fungi tend to rapidly invade agar surfaces, covering small yeast colonies and resulting in an underestimation of their number. In this study, we developed a real-time quantitative PCR (qPCR) method using TaqMan probes to quantify a mixed fungal community containing the most common dairy yeasts and molds: Penicillium camemberti, Geotrichum candidum, Debaryomyces hansenii, and Kluyveromyces lactis on soft-cheese model curds (SCMC). The qPCR method was optimized and validated on pure cultures and used to evaluate the growth dynamics of a ripening culture containing P. camemberti, G. candidum, and K. lactis on the surface of the SCMC during a 31-day ripening period. The results showed that P. camemberti and G. candidum quickly dominated the ecosystem, while K. lactis remained less abundant. When added to this ecosystem, D. hansenii completely inhibited the growth of K. lactis in addition to reducing the growth of the other fungi. This result was confirmed by the decrease in the mycelium biomass on SCMC. This study compares culture-dependent and qPCR methods to successfully quantify complex fungal microbiota on a model curd simulating Camembert-type cheese.

  10. Salt influence on surface microorganisms and ripening of soft ewe cheese.

    PubMed

    Tabla, Rafael; Gómez, Antonia; Rebollo, José E; Roa, Isidro

    2015-05-01

    The effect of different brining treatments on salt uptake and diffusion during the first 30 d of ripening was determined in soft ewe cheese. Additionally, salt influence on surface microorganisms and physicochemical parameters was evaluated. Cheeses were placed into different brine solutions (14, 18 and 24°Bé) at 5 and 10 °C for 1, 2 or 3 h. Samples from rind, outer core and inner core were analysed at 0, 7, 15 and 30 d. Complete salt diffusion from rind to the inner core took about 15 d. The resulting salt gradient favoured the development of a pH gradient from the surface to the inner core. Salt concentration also had a significant effect on the growth of surface microorganisms (mesophiles, pseudomonads and halotolerants). However, mould and yeasts were not affected throughout ripening by the salt levels achieved. Brine salting by immersion for 3 h at 10 °C in 24°B brine was found to be the most suitable treatment to control pseudomonads in cheese rind, as spoilage microorganism.

  11. Determination of optimum harvest maturity and physico-chemical quality of Rastali banana (Musa AAB Rastali) during fruit ripening.

    PubMed

    Kheng, Tee Yei; Ding, Phebe; Abdul Rahman, Nor Aini

    2012-01-15

    A series of physico-chemical quality (peel and pulp colours, pulp firmness, fruit pH, sugars and acids content, respiration rate and ethylene production) were conducted to study the optimum harvest periods (either week 11 or week 12 after emergence of the first hand) of Rastali banana (Musa AAB Rastali) based on the fruit quality during ripening. Rastali banana fruit exhibited a climacteric rise with the peaks of both CO(2) and ethylene production occurring simultaneously at day 3 after ripening was initiated and declined at day 5 when fruits entered the senescence stage. De-greening was observed in both of the harvesting weeks with peel turned from green to yellow, tissue softening, and fruits became more acidic and sweeter as ripening progressed. Sucrose, fructose and glucose were the main sugars found while malic, citric and succinic acids were the main organic acids found in the fruit. Rastali banana harvested at weeks 11 and 12 can be considered as commercial harvest period when the fruits have developed good organoleptic and quality attributes during ripening. However, Rastali banana fruit at more mature stage of harvest maturity taste slightly sweeter and softer with higher ethylene production which also means the fruits may undergo senescence faster than fruit harvested at week 11. Copyright © 2011 Society of Chemical Industry.

  12. Prognostic Factors in Cholinesterase Inhibitor Poisoning.

    PubMed

    Sun, In O; Yoon, Hyun Ju; Lee, Kwang Young

    2015-09-28

    Organophosphates and carbamates are insecticides that are associated with high human mortality. The purpose of this study is to investigate the prognostic factors affecting survival in patients with cholinesterase inhibitor (CI) poisoning. This study included 92 patients with CI poisoning in the period from January 2005 to August 2013. We divided these patients into 2 groups (survivors vs. non-survivors), compared their clinical characteristics, and analyzed the predictors of survival. The mean age of the included patients was 56 years (range, 16-88). The patients included 57 (62%) men and 35 (38%) women. When we compared clinical characteristics between the survivor group (n=81, 88%) and non-survivor group (n=11, 12%), there were no differences in renal function, pancreatic enzymes, or serum cholinesterase level, except for serum bicarbonate level and APACHE II score. The serum bicarbonate level was lower in non-survivors than in survivors (12.45±2.84 vs. 18.36±4.73, P<0.01). The serum APACHE II score was higher in non-survivors than in survivors (24.36±5.22 vs. 12.07±6.67, P<0.01). The development of pneumonia during hospitalization was higher in non-survivors than in survivors (n=9, 82% vs. n=31, 38%, P<0.01). In multiple logistic regression analysis, serum bicarbonate concentration, APACHE II score, and pneumonia during hospitalization were the important prognostic factors in patients with CI poisoning. Serum bicarbonate and APACHE II score are useful prognostic factors in patients with CI poisoning. Furthermore, pneumonia during hospitalization was also important in predicting prognosis in patients with CI poisoning. Therefore, prevention and active treatment of pneumonia is important in the management of patients with CI poisoning.

  13. Suppressing Type 2C Protein Phosphatases Alters Fruit Ripening and the Stress Response in Tomato.

    PubMed

    Zhang, Yushu; Li, Qian; Jiang, Li; Kai, Wenbin; Liang, Bin; Wang, Juan; Du, Yangwei; Zhai, Xiawan; Wang, Jieling; Zhang, Yingqi; Sun, Yufei; Zhang, Lusheng; Leng, Ping

    2018-01-01

    Although ABA signaling has been widely studied in Arabidopsis, the roles of core ABA signaling components in fruit remain poorly understood. Herein, we characterize SlPP2C1, a group A type 2C protein phosphatase that negatively regulates ABA signaling and fruit ripening in tomato. The SlPP2C1 protein was localized in the cytoplasm close to AtAHG3/AtPP2CA. The SlPP2C1 gene was expressed in all tomato tissues throughout development, particularly in flowers and fruits, and it was up-regulated by dehydration and ABA treatment. SlPP2C1 expression in fruits was increased at 30 d after full bloom and peaked at the B + 1 stage. Suppression of SlPP2C1 expression significantly accelerated fruit ripening which was associated with higher levels of ABA signaling genes that are reported to alter the expression of fruit ripening genes involved in ethylene release and cell wall catabolism. SlPP2C1-RNAi (RNA interference) led to increased endogenous ABA accumulation and advanced release of ethylene in transgenic fruits compared with wild-type (WT) fruits. SlPP2C1-RNAi also resulted in abnormal flowers and obstructed the normal abscission of pedicels. SlPP2C1-RNAi plants were hypersensitized to ABA, and displayed delayed seed germination and primary root growth, and increased resistance to drought stress compared with WT plants. These results demonstrated that SlPP2C1 is a functional component in the ABA signaling pathway which participates in fruit ripening, ABA responses and drought tolerance. © The Author 2017. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  14. Recreational Use of Phosphodiesterase 5 Inhibitors and Its Associated Factors among Undergraduate Male Students in an Ethiopian University: A Cross-Sectional Study.

    PubMed

    Gebreyohannes, Eyob Alemayehu; Bhagavathula, Akshaya Srikanth; Gebresillassie, Begashaw Melaku; Tefera, Yonas Getaye; Belachew, Sewunet Admasu; Erku, Daniel Asfaw

    2016-12-01

    To assess the prevalence of phosphodiesterase 5 (PDE5) inhibitor use and associated factors among University of Gondar undergraduate students. An institution-based, cross-sectional study, using a survey questionnaire, was conducted from October to December 2015 to assess PDE5 inhibitor use and associated factors among male students at the University of Gondar. A Self-Esteem and Relationship questionnaire (14 items), an International Index of Erectile Function questionnaire (15 items) and a questionnaire on PDE5 inhibitor use (14 items) were included in the survey. Across all respondents (age, 21.9±1.88 years), more than half (55.7%, n=233) had heard about PDE5 inhibitors, but only 23 men (5.5%) reported trying a PDE5 inhibitor drug at least once. Older students were more likely to use PDE5 inhibitors compared to younger students (adjusted odds ratio [AOR], 1.40; 95% confidence interval [CI], 1.109~1.768). Those students who were smokers were 5.15 times more likely to use PDE5 inhibitors as compared to their non-smoking counterparts (AOR, 5.15; 95% CI, 2.096~12.687). In addition, multivariate logistic regression showed that being in a relationship, alcohol use, greater number of cigarettes smoked per day, and more sexual partners were significantly associated with PDE5 inhibitor use. The prevalence of PDE5 inhibitor use among undergraduate students was 5.5%. Cigarette smoking and other substance use, older age, and greater number of sexual partners were significantly associated factors for PDE5 inhibitor use. These findings suggest that restricting access to PDE5 inhibitor drugs is essential to curtailing misuse among university students.

  15. Evaluating carotenoid changes in tomatoes during postharvest ripening using Raman chemical imaging.

    USDA-ARS?s Scientific Manuscript database

    During the postharvest ripening of tomato fruits, the increasing presence of lycopene in the tomatoe samples spanning a range of fruit maturity. In this study, Raman chemical images were acquired of tomato samples spanning a range of fruit maturity stages, and were evaluated for the presence and di...

  16. Classification of Bitter Orange Essential Oils According to Fruit Ripening Stage by Untargeted Chemical Profiling and Machine Learning.

    PubMed

    Taghadomi-Saberi, Saeedeh; Mas Garcia, Sílvia; Allah Masoumi, Amin; Sadeghi, Morteza; Marco, Santiago

    2018-06-13

    The quality and composition of bitter orange essential oils (EOs) strongly depend on the ripening stage of the citrus fruit. The concentration of volatile compounds and consequently its organoleptic perception varies. While this can be detected by trained humans, we propose an objective approach for assessing the bitter orange from the volatile composition of their EO. The method is based on the combined use of headspace gas chromatography⁻mass spectrometry (HS-GC-MS) and artificial neural networks (ANN) for predictive modeling. Data obtained from the analysis of HS-GC-MS were preprocessed to select relevant peaks in the total ion chromatogram as input features for ANN. Results showed that key volatile compounds have enough predictive power to accurately classify the EO, according to their ripening stage for different applications. A sensitivity analysis detected the key compounds to identify the ripening stage. This study provides a novel strategy for the quality control of bitter orange EO without subjective methods.

  17. Causal Relationship between Microbial Ecology Dynamics and Proteolysis during Manufacture and Ripening of Protected Designation of Origin (PDO) Cheese Canestrato Pugliese

    PubMed Central

    De Pasquale, Ilaria; Calasso, Maria; Mancini, Leonardo; Ercolini, Danilo; La Storia, Antonietta; De Angelis, Maria; Gobbetti, Marco

    2014-01-01

    Pyrosequencing of the 16S rRNA gene, community-level physiological profiles determined by the use of Biolog EcoPlates, and proteolysis analyses were used to characterize Canestrato Pugliese Protected Designation of Origin (PDO) cheese. The number of presumptive mesophilic lactococci in raw ewes' milk was higher than that of presumptive mesophilic lactobacilli. The numbers of these microbial groups increased during ripening, showing temporal and numerical differences. Urea-PAGE showed limited primary proteolysis, whereas the analysis of the pH 4.6-soluble fraction of the cheese revealed that secondary proteolysis increased mainly from 45 to 75 days of ripening. This agreed with the concentration of free amino acids. Raw ewes' milk was contaminated by several bacterial phyla: Proteobacteria (68%; mainly Pseudomonas), Firmicutes (30%; mainly Carnobacterium and Lactococcus), Bacteroidetes (0.05%), and Actinobacteria (0.02%). Almost the same microbial composition persisted in the curd after molding. From day 1 of ripening onwards, the phylum Firmicutes dominated. Lactococcus dominated throughout ripening, and most of the Lactobacillus species appeared only at 7 or 15 days. At 90 days, Lactococcus (87.2%), Lactobacillus (4.8%; mainly Lactobacillus plantarum and Lactobacillus sakei), and Leuconostoc (3.9%) dominated. The relative utilization of carbon sources by the bacterial community reflected the succession. This study identified strategic phases that characterized the manufacture and ripening of Canestrato Pugliese cheese and established a causal relationship between mesophilic lactobacilli and proteolysis. PMID:24771032

  18. Causal relationship between microbial ecology dynamics and proteolysis during manufacture and ripening of protected designation of origin (PDO) cheese Canestrato Pugliese.

    PubMed

    De Pasquale, Ilaria; Calasso, Maria; Mancini, Leonardo; Ercolini, Danilo; La Storia, Antonietta; De Angelis, Maria; Di Cagno, Raffaella; Gobbetti, Marco

    2014-07-01

    Pyrosequencing of the 16S rRNA gene, community-level physiological profiles determined by the use of Biolog EcoPlates, and proteolysis analyses were used to characterize Canestrato Pugliese Protected Designation of Origin (PDO) cheese. The number of presumptive mesophilic lactococci in raw ewes' milk was higher than that of presumptive mesophilic lactobacilli. The numbers of these microbial groups increased during ripening, showing temporal and numerical differences. Urea-PAGE showed limited primary proteolysis, whereas the analysis of the pH 4.6-soluble fraction of the cheese revealed that secondary proteolysis increased mainly from 45 to 75 days of ripening. This agreed with the concentration of free amino acids. Raw ewes' milk was contaminated by several bacterial phyla: Proteobacteria (68%; mainly Pseudomonas), Firmicutes (30%; mainly Carnobacterium and Lactococcus), Bacteroidetes (0.05%), and Actinobacteria (0.02%). Almost the same microbial composition persisted in the curd after molding. From day 1 of ripening onwards, the phylum Firmicutes dominated. Lactococcus dominated throughout ripening, and most of the Lactobacillus species appeared only at 7 or 15 days. At 90 days, Lactococcus (87.2%), Lactobacillus (4.8%; mainly Lactobacillus plantarum and Lactobacillus sakei), and Leuconostoc (3.9%) dominated. The relative utilization of carbon sources by the bacterial community reflected the succession. This study identified strategic phases that characterized the manufacture and ripening of Canestrato Pugliese cheese and established a causal relationship between mesophilic lactobacilli and proteolysis. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  19. The influence of ripening stage and cultivation system on the total antioxidant activity and total phenolic compounds of yellow passion fruit pulp.

    PubMed

    Macoris, Mariana S; De Marchi, Renata; Janzantti, Natália S; Monteiro, Magali

    2012-07-01

    This work aimed to investigate the influence of both ripening stage and cultivation system on the total phenolic compounds (TPC) and total antioxidant activity (TAA) of passion fruit pulp. TPC extraction was optimized using a 2³ central composed design. The variables were fruit pulp volume, methanol volume and extraction solution volume. TPC was determined using the Folin-Ciocalteu reaction, and TAA using the ABTS radical reaction. The conditions to extract TPC were 2 mL passion fruit pulp and 9 mL extraction solution containing 40% methanol:water (v/v). TPC values increased in the passion fruit pulp during ripening for both cultivation systems, ranging from 281.8 to 361.9 mg gallic acid L⁻¹ (P ≤ 0.05) for the organic pulp and from 291.0 to 338.6 mg gallic acid L⁻¹ (P ≤ 0.05) for the conventional pulp. TPC values increased during ripening for both organic and conventional passion fruit. The same was true for TAA values for conventional passion fruit. For organic passion fruit, however, TAA values were highest at the initial ripening stages. These results suggest that antioxidant compounds exert strong influence on the initial ripening stages for organic passion fruit, when TPC still did not reach its maximum level. Copyright © 2012 Society of Chemical Industry.

  20. Isoprenoid, lipid, and protein contents in intact plastids isolated from mesocarp cells of traditional and high-pigment tomato cultivars at different ripening stages.

    PubMed

    Lenucci, Marcello S; Serrone, Lucia; De Caroli, Monica; Fraser, Paul D; Bramley, Peter M; Piro, Gabriella; Dalessandro, Giuseppe

    2012-02-22

    This study reports quali-quantitative analyses on isoprenoids, phospholipids, neutral lipids, phytosterols, and proteins in purified plastids isolated from fresh fruits of traditional (Donald and Incas) and high-pigment (Kalvert and HLY-18) tomato cultivars at four ripening stages. In all of the investigated cultivars, lycopene, β-catotene, lutein, and total carotenoids varied significantly during ripening. Chromoplasts of red-ripe tomato fruits of high-pigment cultivars accumulated twice as much as lycopene (307.6 and 319.2 μg/mg of plastid proteins in Kalvert and HLY-18, respectively) than ordinary cultivars (178.6 and 151.7 μg/mg of plastid proteins in Donald and Incas, respectively); differences in chlorophyll and α-tocopherol contents were also evidenced. Phospholipids and phytosterols increased during ripening, whereas triglycerides showed a general decrease. Regardless of the stage of ripening, palmitic acid was the major fatty acid in all cultivars (ranging from 35 to 52% of the total fatty acids), followed by stearic, oleic, linoleic, linolenic, and myristic acids, but their relative percentage was affected by ripening. Most of the bands detected on the SDS-PAGEs of plastid proteins were constantly present during chloroplast-to-chromoplast conversion, some others disappeared, and only one, with a molecular weight of ~41.6 kDa, was found to increase in intensity.

  1. Modeling of Camembert-type cheese mass loss in a ripening chamber: main biological and physical phenomena.

    PubMed

    Hélias, A; Mirade, P-S; Corrieu, G

    2007-11-01

    A model of the mass loss of Camembert-type cheese was established with data obtained from 2 experimental ripening trials carried out in 2 pilot ripening chambers. During these experiments, a cheese was continuously weighed and the relative humidity, temperature, oxygen, and carbon dioxide concentrations in the ripening chamber were recorded online. The aim was to establish a simple but accurate model that would predict cheese mass changes according to available online measurements. The main hypotheses were that 1) the cheese water activity was constant during ripening, 2) the respiratory activity of the microflora played a major role by inducing heat production, combined with important water evaporation, 3) the temperature gradient existing inside the cheese was negligible, and the limiting phenomenon was the convective transfer. The water activity and the specific heat of the cheeses were assessed by offline measurements. The others parameters in the model were obtained from the literature. This dynamic model was built with 2 state variables: the cheese mass and the surface temperature of the cheese. In this way, only the heat transfer coefficient had to be fitted, and it was strongly determined by the airflow characteristics close to the cheeses. Model efficiency was illustrated by comparing the estimated and measured mass and the mass loss rate for the 2 studied runs; the relative errors were less than 1.9 and 3.2% for the mass loss and the mass loss rate, respectively. The dynamic effects of special events, such as room defrosting or changes in chamber relative humidity, were well described by the model, especially in terms of kinetics (mass loss rates).

  2. Controlled water deficit during ripening affects proanthocyanidin synthesis, concentration and composition in Cabernet Sauvignon grape skins.

    PubMed

    Cáceres-Mella, Alejandro; Talaverano, M Inmaculada; Villalobos-González, Luis; Ribalta-Pizarro, Camila; Pastenes, Claudio

    2017-08-01

    The influence of controlled water deficit on the phenolic composition and gene expression of VvLAR2, VvMYBPA1, VvMYBPA2 and VvMYB4a in Cabernet Sauvignon grape skins throughout ripening was investigated. The assay was carried out on own-rooted Vitis vinifera plants cv. Cabernet Sauvignon in a commercial vineyard from veraison until commercial harvest. Three irrigation regimes were used from veraison until harvest with the following treatments: T1: 3.6 mm day -1 ; T2: 1.8 mm day -1 and T3: 0.3 mm day -1 . The content of total phenols and total anthocyanins in grape skins increased during ripening, but water deficit did not produce differences among treatments in the total anthocyanin concentration. Proanthocyanidins (PAs) decreased throughout ripening, although approximately 25 days after veraison (DAV), their content slightly increased. This effect was more pronounced in the most restrictive treatment (T3). A similar pattern was observed in the transcript abundance of VvLAR2, VvMYBPA1 and VvMYB4a. PAs separation revealed differences in concentration but not in the proportion among fractions among the irrigation treatments. Additionally, controlled water deficit increased the mean degree of polymerization and the flavan-3-ol polymeric concentration in grape skins throughout ripening but with no effects on the extent of PAs galloylation. Our results suggest that the water status of Cabernet Sauvignon grapevines affects the gene expression for proteins involved in the synthesis of PAs, increasing their concentration and also their composition, with further evidence for the efficacy of a convenient, controlled water deficit strategy for grapevine cultivation. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  3. Isolation of genes differentially expressed during development and ripening of Fragaria chiloensis fruit by suppression subtractive hybridization.

    PubMed

    Pimentel, Paula; Salvatierra, Ariel; Moya-León, María Alejandra; Herrera, Raúl

    2010-09-15

    Fragaria chiloensis, the native Chilean strawberry, is noted for its good fruit quality characters. However, it is a highly perishable fruit due to its rapid softening. With the aim to screen for genes differentially expressed during development and ripening of strawberry fruit, the subtractive suppressive hybridization (SSH) methodology was employed. Six libraries were generated contrasting transcripts from four different developmental stages. A set of 1807 genes was isolated and characterized. In our EST collection, approximately 90% of partial cDNAs showed significant similarity to proteins with known or unknown function registered in databases. Among them, proteins related to protein fate were identified in a large green fruit library and protein related with cellular transport, cell wall-related proteins, and transcription regulators were identified in a ripe fruit library. Thirteen genes were analyzed by qRT-PCR during development and ripening of the Chilean strawberry fruit. The information generated in this study provides new clues to aid the understanding of the ripening process in F. chiloensis fruit. Copyright 2010 Elsevier GmbH. All rights reserved.

  4. Asynchronous ripening behavior of cactus pear (Opuntia ficus-indica) cultivars with respect to physicochemical and physiological attributes.

    PubMed

    Kyriacou, M C; Emmanouilidou, M G; Soteriou, G A

    2016-11-15

    Physicochemical and physiological ripening events in cactus pear (Opuntia ficus-indica) fruit of cultivars 'Ntopia' and 'Hercules' were profiled against skin coloration from mature-green (S1) to over-mature (S5). Fructose and glucose accumulation were linear in 'Ntopia' but peaked near S3 in 'Hercules' synchronously to the appearance of sucrose. Betalains increased steadily in 'Ntopia' (103.2mg/l) but peaked before full skin coloration in 'Hercules' (49.7mg/l); whereas phenolic content remained invariable and ascorbate content peaked near S5 in both 'Ntopia' (108.6μg/g) and 'Hercules' (163.1μg/g). Cell wall material diminished with maturity though textural changes with ripening appeared not related to pectin solubilization but to weakening of glycan bonding and loss of neutral sugars. Fruit firmness rather was correlated to seed weight (r=0.89) and seed-to-pulp ratio (r=0.73). Cultivar differences highlighted in the chronology of ripening events are critical for defining optimum harvest maturity and postharvest handling protocols for premium quality cactus pear fruit. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Skin problems and EGFR-tyrosine kinase inhibitor

    PubMed Central

    Kozuki, Toshiyuki

    2016-01-01

    Epidermal growth factor receptor inhibition is a good target for the treatment of lung, colon, pancreatic and head and neck cancers. Epidermal growth factor receptor-tyrosine kinase inhibitor was first approved for the treatment of advanced lung cancer in 2002. Epidermal growth factor receptor-tyrosine kinase inhibitor plays an essential role in the treatment of cancer, especially for patients harbouring epidermal growth factor receptor activating mutation. Hence, skin toxicity is the most concerning issue for the epidermal growth factor receptor-tyrosine kinase inhibitor treatment. Skin toxicity is bothersome and sometimes affects the quality of life and treatment compliance. Thus, it is important for physicians to understand the background and how to manage epidermal growth factor receptor-tyrosine kinase inhibitor-associated skin toxicity. Here, the author reviewed the mechanism and upfront preventive and reactive treatments for epidermal growth factor receptor inhibitor-associated skin toxicities. PMID:26826719

  6. The onset of grapevine berry ripening is characterized by ROS accumulation and lipoxygenase-mediated membrane peroxidation in the skin

    PubMed Central

    2014-01-01

    Background The ripening of fleshy fruits is a complex developmental program characterized by extensive transcriptomic and metabolic remodeling in the pericarp tissues (pulp and skin) making unripe green fruits soft, tasteful and colored. The onset of ripening is regulated by a plethora of endogenous signals tuned to external stimuli. In grapevine and tomato, which are classified as non-climacteric and climacteric species respectively, the accumulation of hydrogen peroxide (H2O2) and extensive modulation of reactive oxygen species (ROS) scavenging enzymes at the onset of ripening has been reported, suggesting that ROS could participate to the regulatory network of fruit development. In order to investigate this hypothesis, a comprehensive biochemical study of the oxidative events occurring at the beginning of ripening in Vitis vinifera cv. Pinot Noir has been undertaken. Results ROS-specific staining allowed to visualize not only H2O2 but also singlet oxygen (1O2) in berry skin cells just before color change in distinct subcellular locations, i.e. cytosol and plastids. H2O2 peak in sample skins at véraison was confirmed by in vitro quantification and was supported by the concomitant increase of catalase activity. Membrane peroxidation was also observed by HPLC-MS on galactolipid species at véraison. Mono- and digalactosyl diacylglycerols were found peroxidized on one or both α-linolenic fatty acid chains, with a 13(S) absolute configuration implying the action of a specific enzyme. A lipoxygenase (PnLOXA), expressed at véraison and localizing inside the chloroplasts, was indeed able to catalyze membrane galactolipid peroxidation when overexpressed in tobacco leaves. Conclusions The present work demonstrates the controlled, harmless accumulation of specific ROS in distinct cellular compartments, i.e. cytosol and chloroplasts, at a definite developmental stage, the onset of grape berry ripening. These features strongly candidate ROS as cellular signals in fruit

  7. Reduction of biogenic amines in sufu by ethanol addition during ripening stage

    USDA-ARS?s Scientific Manuscript database

    The aim of this study was to investigate the contents of biogenic amines (BAs) in different types of sufu samples obtained from different producers, and also the possible effect of addition of ethanol on reducing the levels of BAs during sufu ripening. The levels of BAs in sufu samples were analyze...

  8. Secretion imbalance between tumour necrosis factor and its inhibitor in inflammatory bowel disease

    PubMed Central

    Noguchi, M; Hiwatashi, N; Liu, Z; Toyota, T

    1998-01-01

    Background—Tumour necrosis factor (TNF) α and TNF-β are soluble ligands binding to TNF receptors with similar activities; soluble TNF receptors neutralise TNF activity by acting as inhibitors. Little is known about the cytokine/soluble receptor role in inflammatory bowel disease (IBD). 
Aims—To test the hypothesis that an imbalance in secretion between TNF and TNF inhibitors plays a role in gut inflammation in patients with IBD. 
Methods—The secretion of TNF-α, TNF-β, and soluble TNF receptors was compared in the culture supernatants of colonic biopsy specimens and isolated lamina propria mononuclear cells from patients with active colonic IBD. 
Results—Spontaneous secretion of TNF-α in involved IBD mucosa was higher than in normal control and self limited colitis mucosa. Secretion of TNF-β was higher in patients with Crohn's disease than in those with ulcerative colitis. Soluble TNF receptor in IBD mucosa inhibited TNF activity. Type 2 soluble receptor release from IBD mucosa was increased in active inflammation; release from lamina propria cells was not increased. Mucosal TNF-α production correlated with severity of disease. 
Conclusions—Results showed enhanced secretion of TNF-α but failure to release enhanced amounts of soluble TNF receptor in lamina propria mononuclear cells of patients with IBD. An imbalance in secretion between TNF and TNF inhibitor may be implicated in the pathogenesis of IBD. 

 Keywords: Crohn's disease; inflammation; mucosal immunology; soluble TNF receptor; tumour necrosis factor; ulcerative colitis PMID:10189845

  9. Energetic factors determining the binding of type I inhibitors to c-Met kinase: experimental studies and quantum mechanical calculations

    PubMed Central

    Yu, Zhe; Ma, Yu-chi; Ai, Jing; Chen, Dan-qi; Zhao, Dong-mei; Wang, Xin; Chen, Yue-lei; Geng, Mei-yu; Xiong, Bing; Cheng, Mao-sheng; Shen, Jing-kang

    2013-01-01

    Aim: To decipher the molecular interactions between c-Met and its type I inhibitors and to facilitate the design of novel c-Met inhibitors. Methods: Based on the prototype model inhibitor 1, four ligands with subtle differences in the fused aromatic rings were synthesized. Quantum chemistry was employed to calculate the binding free energy for each ligand. Symmetry-adapted perturbation theory (SAPT) was used to decompose the binding energy into several fundamental forces to elucidate the determinant factors. Results: Binding free energies calculated from quantum chemistry were correlated well with experimental data. SAPT calculations showed that the predominant driving force for binding was derived from a sandwich π–π interaction with Tyr-1230. Arg-1208 was the differentiating factor, interacting with the 6-position of the fused aromatic ring system through the backbone carbonyl with a force pattern similar to hydrogen bonding. Therefore, a hydrogen atom must be attached at the 6-position, and changing the carbon atom to nitrogen caused unfavorable electrostatic interactions. Conclusion: The theoretical studies have elucidated the determinant factors involved in the binding of type I inhibitors to c-Met. PMID:24056705

  10. Inhibitor development in patients receiving recombinant factor VIII (Recombinate rAHF/Bioclate): a prospective pharmacovigilance study.

    PubMed

    Ewenstein, B M; Gomperts, E D; Pearson, S; O'Banion, M E

    2004-09-01

    Clinical trials to date have not been adequately powered to assess comparatively infrequent events such as inhibitor development in previously treated patients (PTPs). Comprehensive large-scale pharmacovigilance studies can be useful for this purpose. We prospectively collected inhibitor development reports worldwide among recipients of Recombinate rAHF recombinant factor VIII (rFVIII), also formerly distributed under the product name Bioclate, for the entire postlicensure period from 1993 through 2002. To determine level of exposure to rFVIII we also compiled the Recombinate rAHF/Bioclate International Units (IU) distributed annually. To estimate inhibitor incidence separately for previously untreated or minimally treated patients (PUPs) with 1-50 exposure days and PTPs with >50 exposure days, we used haemophilia A incidence and prevalence data and pooled mean annual rFVIII consumption per PUP and PTP from international multicentre prospective clinical trials. Documented inhibitor cases totalled 89, and the total quantity of Recombinate rAHF/Bioclate rFVIII distributed was 6.48 x10(9) IU. No lot association or other clustering of inhibitor events was evident in PTPs. The incidence of all reported inhibitors, expressed as a percentage of patients treated, was 11.9% (CI: 5.05-28.0%) for PUPs when compared with 0.123% (CI: 0.030-0.512%) for PTPs. The rates for high-titre inhibitors (>5 BU) only were 5.96% (CI: 3.00-11.8%) for PUPs and 0.0554% (CI: 0.0113-0.271%) for PTPs. Thus, incidence rates for both all inhibitors and high-titre inhibitors in PTPs were 1% of the corresponding rates in PUPs. Data from prospective PUP clinical trials involving intensive active monitoring suggest that true inhibitor incidence may be approximately twice that estimated in this pharmacovigilance study. Nevertheless, inhibitor development in PTPs receiving Recombinate rAHF/Bioclate is infrequent.

  11. Evolution of the composition of a selected bitter Camembert cheese during ripening: release and migration of taste-active compounds.

    PubMed

    Engel, E; Tournier, C; Salles, C; Le Quéré, J L

    2001-06-01

    The aim of this study was to add to the understanding of changes in taste that occur during the ripening of a bitter Camembert cheese by the evolution of its composition. Physicochemical analyses were performed on rind, under-rind, and center portions of a Camembert cheese selected for its intense bitterness. At each of the six steps of ripening studied organic acids, sugars, total nitrogen, soluble nitrogen, phosphotungstic acid soluble nitrogen, non-protein nitrogen, Na, K, Ca, Mg, Pi, Cl, and biogenic amines were quantified in each portion. Changes in cheese composition seemed to mainly result from the development of Penicillium camemberti on the cheese outer layer. Migration phenomena and the release of potentially taste-active compounds allowed for the evolution of saltiness, sourness, and bitterness throughout ripening to be better understood. Apart from taste-active compounds, the impact of the cheese matrix on its taste development is discussed.

  12. Recreational Use of Phosphodiesterase 5 Inhibitors and Its Associated Factors among Undergraduate Male Students in an Ethiopian University: A Cross-Sectional Study

    PubMed Central

    Bhagavathula, Akshaya Srikanth; Gebresillassie, Begashaw Melaku; Tefera, Yonas Getaye; Belachew, Sewunet Admasu; Erku, Daniel Asfaw

    2016-01-01

    Purpose To assess the prevalence of phosphodiesterase 5 (PDE5) inhibitor use and associated factors among University of Gondar undergraduate students. Materials and Methods An institution-based, cross-sectional study, using a survey questionnaire, was conducted from October to December 2015 to assess PDE5 inhibitor use and associated factors among male students at the University of Gondar. A Self-Esteem and Relationship questionnaire (14 items), an International Index of Erectile Function questionnaire (15 items) and a questionnaire on PDE5 inhibitor use (14 items) were included in the survey. Results Across all respondents (age, 21.9±1.88 years), more than half (55.7%, n=233) had heard about PDE5 inhibitors, but only 23 men (5.5%) reported trying a PDE5 inhibitor drug at least once. Older students were more likely to use PDE5 inhibitors compared to younger students (adjusted odds ratio [AOR], 1.40; 95% confidence interval [CI], 1.109~1.768). Those students who were smokers were 5.15 times more likely to use PDE5 inhibitors as compared to their non-smoking counterparts (AOR, 5.15; 95% CI, 2.096~12.687). In addition, multivariate logistic regression showed that being in a relationship, alcohol use, greater number of cigarettes smoked per day, and more sexual partners were significantly associated with PDE5 inhibitor use. Conclusions The prevalence of PDE5 inhibitor use among undergraduate students was 5.5%. Cigarette smoking and other substance use, older age, and greater number of sexual partners were significantly associated factors for PDE5 inhibitor use. These findings suggest that restricting access to PDE5 inhibitor drugs is essential to curtailing misuse among university students. PMID:28053948

  13. Surface microbial consortia from Livarot, a French smear-ripened cheese.

    PubMed

    Larpin-Laborde, Sandra; Imran, Muhammad; Bonaïti, Catherine; Bora, Nagamani; Gelsomino, Roberto; Goerges, Stefanie; Irlinger, Françoise; Goodfellow, Michael; Ward, Alan C; Vancanneyt, Marc; Swings, Jean; Scherer, Siegfried; Guéguen, Micheline; Desmasures, Nathalie

    2011-08-01

    The surface microflora (902 isolates) of Livarot cheeses from three dairies was investigated during ripening. Yeasts were mainly identified by Fourier transform infrared spectroscopy. Geotrichum candidum was the dominating yeast among 10 species. Bacteria were identified using Biotype 100 strips, dereplicated by repetitive extragenic palindromic PCR (rep-PCR); 156 representative strains were identified by either BOX-PCR or (GTG)(5)-PCR, and when appropriate by 16S rDNA sequencing and SDS-PAGE analysis. Gram-positive bacteria accounted for 65% of the isolates and were mainly assigned to the genera Arthrobacter , Brevibacterium , Corynebacterium , and Staphylococcus . New taxa related to the genera Agrococcus and Leucobacter were found. Yeast and Gram-positive bacteria strains deliberately added as smearing agents were sometimes undetected during ripening. Thirty-two percent of the isolates were Gram-negative bacteria, which showed a high level of diversity and mainly included members of the genera Alcaligenes , Hafnia , Proteus , Pseudomonas , and Psychrobacter . Whatever the milk used (pasteurized or unpasteurized), similar levels of biodiversity were observed in the three dairies, all of which had efficient cleaning procedures and good manufacturing practices. It appears that some of the Gram-negative bacteria identified should now be regarded as potentially useful in some cheese technologies. The assessment of their positive versus negative role should be objectively examined.

  14. The Composition of Camembert Cheese-Ripening Cultures Modulates both Mycelial Growth and Appearance

    PubMed Central

    Lessard, Marie-Hélène; Bélanger, Gaétan; St-Gelais, Daniel

    2012-01-01

    The fungal microbiota of bloomy-rind cheeses, such as Camembert, forms a complex ecosystem that has not been well studied, and its monitoring during the ripening period remains a challenge. One limitation of enumerating yeasts and molds on traditional agar media is that hyphae are multicellular structures, and colonies on a petri dish rarely develop from single cells. In addition, fungi tend to rapidly invade agar surfaces, covering small yeast colonies and resulting in an underestimation of their number. In this study, we developed a real-time quantitative PCR (qPCR) method using TaqMan probes to quantify a mixed fungal community containing the most common dairy yeasts and molds: Penicillium camemberti, Geotrichum candidum, Debaryomyces hansenii, and Kluyveromyces lactis on soft-cheese model curds (SCMC). The qPCR method was optimized and validated on pure cultures and used to evaluate the growth dynamics of a ripening culture containing P. camemberti, G. candidum, and K. lactis on the surface of the SCMC during a 31-day ripening period. The results showed that P. camemberti and G. candidum quickly dominated the ecosystem, while K. lactis remained less abundant. When added to this ecosystem, D. hansenii completely inhibited the growth of K. lactis in addition to reducing the growth of the other fungi. This result was confirmed by the decrease in the mycelium biomass on SCMC. This study compares culture-dependent and qPCR methods to successfully quantify complex fungal microbiota on a model curd simulating Camembert-type cheese. PMID:22247164

  15. Survival of cheese-ripening microorganisms in a dynamic simulator of the gastrointestinal tract.

    PubMed

    Adouard, Nadège; Magne, Laurent; Cattenoz, Thomas; Guillemin, Hervé; Foligné, Benoît; Picque, Daniel; Bonnarme, Pascal

    2016-02-01

    A mixture of nine microorganisms (six bacteria and three yeasts) from the microflora of surface-ripened cheeses were subjected to in vitro digestive stress in a three-compartment "dynamic gastrointestinal digester" (DIDGI). We studied the microorganisms (i) grown separately in culture medium only (ii) grown separately in culture medium and then mixed, (iii) grown separately in culture medium and then included in a rennet gel and (iv) grown together in smear-ripened cheese. The yeasts Geotrichum candidum, Kluyveromyces lactis and Debaryomyces hansenii, were strongly resistant to the whole DIDGI process (with a drop in viable cell counts of less than <1 log CFU mL(-1)) and there were no significant differences between lab cultures and cheese-grown cultures. Ripening bacteria such as Hafnia alvei survived gastric stress less well when grown in cheese (with no viable cells after 90 min of exposure of the cheese matrix, compared with 6 CFU mL(-1) in lab cultures). The ability of Corynebacterium casei and Staphylococcus equorum to withstand digestive stress was similar for cheese and pure culture conditions. When grow in a cheese matrix, Brevibacterium aurantiacum and Arthrobacter arilaitensis were clearly more sensitive to the overall digestive process than when grown in pure cultures. Lactococcus lactis displayed poorer survival in gastric and duodenal compartments when it had been grown in cheese. In vivo experiments in BALB/c mice agreed with the DIDGI experiments and confirmed the latter's reliability. Copyright © 2015. Published by Elsevier Ltd.

  16. Genome-Wide Association Mapping of Flowering and Ripening Periods in Apple

    PubMed Central

    Urrestarazu, Jorge; Muranty, Hélène; Denancé, Caroline; Leforestier, Diane; Ravon, Elisa; Guyader, Arnaud; Guisnel, Rémi; Feugey, Laurence; Aubourg, Sébastien; Celton, Jean-Marc; Daccord, Nicolas; Dondini, Luca; Gregori, Roberto; Lateur, Marc; Houben, Patrick; Ordidge, Matthew; Paprstein, Frantisek; Sedlak, Jiri; Nybom, Hilde; Garkava-Gustavsson, Larisa; Troggio, Michela; Bianco, Luca; Velasco, Riccardo; Poncet, Charles; Théron, Anthony; Moriya, Shigeki; Bink, Marco C. A. M.; Laurens, François; Tartarini, Stefano; Durel, Charles-Eric

    2017-01-01

    Deciphering the genetic control of flowering and ripening periods in apple is essential for breeding cultivars adapted to their growing environments. We implemented a large Genome-Wide Association Study (GWAS) at the European level using an association panel of 1,168 different apple genotypes distributed over six locations and phenotyped for these phenological traits. The panel was genotyped at a high-density of SNPs using the Axiom®Apple 480 K SNP array. We ran GWAS with a multi-locus mixed model (MLMM), which handles the putatively confounding effect of significant SNPs elsewhere on the genome. Genomic regions were further investigated to reveal candidate genes responsible for the phenotypic variation. At the whole population level, GWAS retained two SNPs as cofactors on chromosome 9 for flowering period, and six for ripening period (four on chromosome 3, one on chromosome 10 and one on chromosome 16) which, together accounted for 8.9 and 17.2% of the phenotypic variance, respectively. For both traits, SNPs in weak linkage disequilibrium were detected nearby, thus suggesting the existence of allelic heterogeneity. The geographic origins and relationships of apple cultivars accounted for large parts of the phenotypic variation. Variation in genotypic frequency of the SNPs associated with the two traits was connected to the geographic origin of the genotypes (grouped as North+East, West and South Europe), and indicated differential selection in different growing environments. Genes encoding transcription factors containing either NAC or MADS domains were identified as major candidates within the small confidence intervals computed for the associated genomic regions. A strong microsynteny between apple and peach was revealed in all the four confidence interval regions. This study shows how association genetics can unravel the genetic control of important horticultural traits in apple, as well as reduce the confidence intervals of the associated regions identified

  17. Genome-Wide Association Mapping of Flowering and Ripening Periods in Apple.

    PubMed

    Urrestarazu, Jorge; Muranty, Hélène; Denancé, Caroline; Leforestier, Diane; Ravon, Elisa; Guyader, Arnaud; Guisnel, Rémi; Feugey, Laurence; Aubourg, Sébastien; Celton, Jean-Marc; Daccord, Nicolas; Dondini, Luca; Gregori, Roberto; Lateur, Marc; Houben, Patrick; Ordidge, Matthew; Paprstein, Frantisek; Sedlak, Jiri; Nybom, Hilde; Garkava-Gustavsson, Larisa; Troggio, Michela; Bianco, Luca; Velasco, Riccardo; Poncet, Charles; Théron, Anthony; Moriya, Shigeki; Bink, Marco C A M; Laurens, François; Tartarini, Stefano; Durel, Charles-Eric

    2017-01-01

    Deciphering the genetic control of flowering and ripening periods in apple is essential for breeding cultivars adapted to their growing environments. We implemented a large Genome-Wide Association Study (GWAS) at the European level using an association panel of 1,168 different apple genotypes distributed over six locations and phenotyped for these phenological traits. The panel was genotyped at a high-density of SNPs using the Axiom®Apple 480 K SNP array. We ran GWAS with a multi-locus mixed model (MLMM), which handles the putatively confounding effect of significant SNPs elsewhere on the genome. Genomic regions were further investigated to reveal candidate genes responsible for the phenotypic variation. At the whole population level, GWAS retained two SNPs as cofactors on chromosome 9 for flowering period, and six for ripening period (four on chromosome 3, one on chromosome 10 and one on chromosome 16) which, together accounted for 8.9 and 17.2% of the phenotypic variance, respectively. For both traits, SNPs in weak linkage disequilibrium were detected nearby, thus suggesting the existence of allelic heterogeneity. The geographic origins and relationships of apple cultivars accounted for large parts of the phenotypic variation. Variation in genotypic frequency of the SNPs associated with the two traits was connected to the geographic origin of the genotypes (grouped as North+East, West and South Europe), and indicated differential selection in different growing environments. Genes encoding transcription factors containing either NAC or MADS domains were identified as major candidates within the small confidence intervals computed for the associated genomic regions. A strong microsynteny between apple and peach was revealed in all the four confidence interval regions. This study shows how association genetics can unravel the genetic control of important horticultural traits in apple, as well as reduce the confidence intervals of the associated regions identified

  18. The auxin response factor gene family in banana: genome-wide identification and expression analyses during development, ripening, and abiotic stress

    PubMed Central

    Hu, Wei; Zuo, Jiao; Hou, Xiaowan; Yan, Yan; Wei, Yunxie; Liu, Juhua; Li, Meiying; Xu, Biyu; Jin, Zhiqiang

    2015-01-01

    Auxin signaling regulates various auxin-responsive genes via two types of transcriptional regulators, Auxin Response Factors (ARF) and Aux/IAA. ARF transcription factors act as critical components of auxin signaling that play important roles in modulating various biological processes. However, limited information about this gene family in fruit crops is currently available. Herein, 47 ARF genes were identified in banana based on its genome sequence. Phylogenetic analysis of the ARFs from banana, rice, and Arabidopsis suggested that the ARFs could be divided into four subgroups, among which most ARFs from the banana showed a closer relationship with those from rice than those from Arabidopsis. Conserved motif analysis showed that all identified MaARFs had typical DNA-binding and ARF domains, but 12 members lacked the dimerization domain. Gene structure analysis showed that the number of exons in MaARF genes ranged from 5 to 21, suggesting large variation amongst banana ARF genes. The comprehensive expression profiles of MaARF genes yielded useful information about their involvement in diverse tissues, different stages of fruit development and ripening, and responses to abiotic stresses in different varieties. Interaction networks and co-expression assays indicated the strong transcriptional response of banana ARFs and ARF-mediated networks in early fruit development for different varieties. Our systematic analysis of MaARFs revealed robust tissue-specific, development-dependent, and abiotic stress-responsive candidate MaARF genes for further functional assays in planta. These findings could lead to potential applications in the genetic improvement of banana cultivars, and yield new insights into the complexity of the control of MaARF gene expression at the transcriptional level. Finally, they support the hypothesis that ARFs are a crucial component of the auxin signaling pathway, which regulates a wide range of physiological processes. PMID:26442055

  19. Covalent cross-linking of insulin-like growth factor-1 to a specific inhibitor from human serum

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ooi, G.T.; Herington, A.C.

    1986-05-29

    Previous studies have shown that a specific inhibitor of insulin-like growth factor (IGF) action in vitro can be isolated from normal human serum and subsequently partially purified on an IGF-affinity column. The ability of the inhibitor to bind the IGFs has now been confirmed directly using covalent cross-linking techniques. When /sup 125/I-IGF-1 was cross-linked to inhibitor using disuccinimidyl suberate, five specifically labelled bands were seen on SDS-PAGE and autoradiography. Two bands (MW 21.5 K and 25.5 K) were intensely labelled, while the remaining three (MW 37 K, 34 K and 18 K) appeared as minor bands only. Inhibitor bioactivity, followingmore » further analysis by hydrophobic interaction chromatography or Con A-Sepharose affinity chromatography, was always associated with the presence of the 21.5 K and/or 25.5 K bands.« less

  20. Comparison of Clinical Factors Between Patients With Angiotensin-Converting Enzyme Inhibitor-Induced Angioedema and Cough.

    PubMed

    Hallberg, Pär; Nagy, Julia; Karawajczyk, Malgorzata; Nordang, Leif; Islander, Gunilla; Norling, Pia; Johansson, Hans-Erik; Kämpe, Mary; Hugosson, Svante; Yue, Qun-Ying; Wadelius, Mia

    2017-04-01

    Angioedema is a rare and serious adverse drug reaction (ADR) to angiotensin-converting enzyme (ACE) inhibitor treatment. Dry cough is a common side effect of ACE inhibitors and has been identified as a possible risk factor for angioedema. We compared characteristics between patients with ACE inhibitor-induced angioedema and cough with the aim of identifying risk factors that differ between these adverse events. Data on patients with angioedema or cough induced by ACE inhibitors were collected from the Swedish database of spontaneously reported ADRs or from collaborating clinicians. Wilcoxon rank sum test, Fisher's exact test, and odds ratios (ORs) with 95% CIs were used to test for between-group differences. The significance threshold was set to P <0.00128 to correct for multiple comparisons. Clinical characteristics were compared between 168 patients with angioedema and 121 with cough only. Smoking and concomitant selective calcium channel blocker treatment were more frequent among patients with angioedema than cough: OR = 4.3, 95% CI = 2.1-8.9, P = 2.2 × 10 -5 , and OR = 3.7, 95% CI = 2.0-7.0, P = 1.7 × 10 -5 . Angioedema cases were seen more often in male patients (OR = 2.2, 95% CI = 1.4-3.6, P = 1.3 × 10 -4 ) and had longer time to onset and higher doses than those with cough ( P = 3.2 × 10 -10 and P = 2.6 × 10 -4 ). A multiple model containing the variables smoking, concurrent calcium channel blocker treatment, male sex, and time to onset accounted for 26% of the variance between the groups. Smoking, comedication with selective calcium channel blockers, male sex, and longer treatment time were associated with ACE inhibitor-induced angioedema rather than cough.

  1. Toward the integration of expert knowledge and instrumental data to control food processes: application to Camembert-type cheese ripening.

    PubMed

    Sicard, M; Perrot, N; Leclercq-Perlat, M-N; Baudrit, C; Corrieu, G

    2011-01-01

    Modeling the cheese ripening process remains a challenge because of its complexity. We still lack the knowledge necessary to understand the interactions that take place at different levels of scale during the process. However, information may be gathered from expert knowledge. Combining this expertise with knowledge extracted from experimental databases may allow a better understanding of the entire ripening process. The aim of this study was to elicit expert knowledge and to check its validity to assess the evolution of organoleptic quality during a dynamic food process: Camembert cheese ripening. Experiments on a pilot scale were carried out at different temperatures and relative humidities to obtain contrasting ripening kinetics. During these experiments, macroscopic evolution was evaluated from an expert's point of view and instrumental measurements were carried out to simultaneously monitor microbiological, physicochemical, and biochemical kinetics. A correlation of 76% was established between the microbiological, physicochemical, and biochemical data and the sensory phases measured according to expert knowledge, highlighting the validity of the experts' measurements. In the future, it is hoped that this expert knowledge may be integrated into food process models to build better decision-aid systems that will make it possible to preserve organoleptic qualities by linking them to other phenomena at the microscopic level. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  2. Skin problems and EGFR-tyrosine kinase inhibitor.

    PubMed

    Kozuki, Toshiyuki

    2016-04-01

    Epidermal growth factor receptor inhibition is a good target for the treatment of lung, colon, pancreatic and head and neck cancers. Epidermal growth factor receptor-tyrosine kinase inhibitor was first approved for the treatment of advanced lung cancer in 2002. Epidermal growth factor receptor-tyrosine kinase inhibitor plays an essential role in the treatment of cancer, especially for patients harbouring epidermal growth factor receptor activating mutation. Hence, skin toxicity is the most concerning issue for the epidermal growth factor receptor-tyrosine kinase inhibitor treatment. Skin toxicity is bothersome and sometimes affects the quality of life and treatment compliance. Thus, it is important for physicians to understand the background and how to manage epidermal growth factor receptor-tyrosine kinase inhibitor-associated skin toxicity. Here, the author reviewed the mechanism and upfront preventive and reactive treatments for epidermal growth factor receptor inhibitor-associated skin toxicities. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  3. Phenol metabolism in the leaves of the olive tree (Olea europaea L.) cv. Picual, Verdial, Arbequina, and Frantoio during ripening.

    PubMed

    Ortega-García, Francisca; Peragón, Juan

    2010-12-08

    The kinetic behavior and protein-expression level of phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO) have been determined in the leaves of the olive tree (Olea europaea L.) of cv. Picual, Verdial, Arbequina, and Frantoio during fruit ripening. Moreover, the concentration of total phenolic compounds, oleuropein, hydroxytyrosol, and tyrosol has been also determined. This study was carried out in 20-year-old olive trees grown in Jaén (Spain). The concentration of total and specific phenols showed a specific pattern in each cultivar. Frantoio showed the highest phenol concentration followed by Arbequina, Picual, and Verdial. A coordinated response between PAL, PPO, and the concentration of total phenols in the four cultivars was found. Also, specific changes were shown over the course of ripening, indicating a regulation of PAL, PPO, and phenol concentration in the olive-tree leaves during fruit ripening.

  4. iTRAQ-based protein profiling provides insights into the central metabolism changes driving grape berry development and ripening

    PubMed Central

    2013-01-01

    Background Grapevine (Vitis vinifera L.) is an economically important fruit crop. Quality-determining grape components such as sugars, acids, flavors, anthocyanins, tannins, etc., accumulate in the different grape berry development stages. Thus, correlating the proteomic profiles with the biochemical and physiological changes occurring in grape is of paramount importance to advance in our understanding of berry development and ripening processes. Results We report the developmental analysis of Vitis vinifera cv. Muscat Hamburg berries at the protein level from fruit set to full ripening. An iTRAQ-based bottom-up proteomic approach followed by tandem mass spectrometry led to the identification and quantitation of 411 and 630 proteins in the green and ripening phases, respectively. Two key points in development relating to changes in protein level were detected: end of the first growth period (7 mm-to-15 mm) and onset of ripening (15 mm-to-V100, V100-to-110). A functional analysis was performed using the Blast2GO software based on the enrichment of GO terms during berry growth. Conclusions The study of the proteome contributes to decipher the biological processes and metabolic pathways involved in the development and quality traits of fruit and its derived products. These findings lie mainly in metabolism and storage of sugars and malate, energy-related pathways such as respiration, photosynthesis and fermentation, and the synthesis of polyphenolics as major secondary metabolites in grape berry. In addition, some key steps in carbohydrate and malate metabolism have been identified in this study, i.e., PFP-PFK or SuSy-INV switches among others, which may influence the final sugar and acid balance in ripe fruit. In conclusion, some proteins not reported to date have been detected to be deregulated in specific tissues and developmental stages, leading to formulate new hypotheses on the metabolic processes underlying grape berry development. These results open up

  5. Biogenic amines in Zamorano cheese: factors involved in their accumulation.

    PubMed

    Combarros-Fuertes, Patricia; Fernández, Domingo; Arenas, Ricardo; Diezhandino, Isabel; Tornadijo, Maria Eugenia; Fresno, José María

    2016-01-15

    Ripened cheese is among fermented food the most often associated with food poisoning from biogenic amines. The influence of ripening time, heat treatment of milk and the effect of using milk from a different ewe breed on the biogenic amine (BA) content of Zamorano cheese was studied by high-performance liquid chromatography. Physicochemical, proteolytic and microbiological parameters were also studied. BA content increased significantly during ripening and their final values were around 400 mg kg(-1). Cheeses elaborated with raw milk duplicated the concentration of BA relative to those elaborated with pasteurized milk (72 °C for 20 s). The average levels of putrescine, spermine and tyramine were higher in cheeses made with a greater proportion of milk from Churra breed. Significant differences in microbial counts and nitrogen soluble in 5% phosphotungstic acid were observed between the different batches. Ripening time and heat treatment applied to milk were the factors that exercised the greatest influence upon the concentration of BA in Zamorano cheese. © 2015 Society of Chemical Industry.

  6. Influence of vegetable coagulant and ripening time on the lipolytic and sensory profile of cheeses made with raw goat milk from Canary breeds.

    PubMed

    Rincón, Arturo A; Pino, Verónica; Fresno, María R; Jiménez-Abizanda, Ana I; Álvarez, Sergio; Ayala, Juan H; Afonso, Ana M

    2017-04-01

    Free fatty acids and sensory profiles were obtained for cheeses made with raw goat milk and vegetable coagulant, derived from the cardoon flower ( Cynara cardunculus), at different ripening times (7 and 20 days). A solid-liquid phase extraction method followed by solid-phase extraction and gas chromatography was used. Profiles were also obtained with cheeses made with commercial coagulant, traditional kid rennet paste, and mixture coagulant (vegetable coagulant-kid rennet). The use of vegetable coagulant and vegetable coagulant-kid rennet is common in traditional Protected Designation of Origin cheeses such as " Queso Flor de Guía" and " Queso Media Flor de Guía" (Spain). Contents of short-chain free fatty acids (7.5-22.5 mmol·kg -1 ), medium-chain free fatty acids (0.4-3.7 mmol·kg -1 ), and long-chain free fatty acids (0.2-2.1 mmol·kg -1 ) varied depending on the coagulant type and the ripening time. Vegetable coagulant cheeses present odour intensity and flavour intensity much higher than commercial coagulant cheeses in the sensory analysis for cheeses obtained with seven days of ripening, but the values decrease when increasing the ripening time. Multivariate analysis allowed us to differentiate cheese samples according to the ripening time when using lipolytic profile and according to the coagulant type using the sensory profile.

  7. Chlorophyll metabolism in pollinated vs. parthenocarpic fig fruits throughout development and ripening.

    PubMed

    Rosianskey, Yogev; Dahan, Yardena; Yadav, Sharawan; Freiman, Zohar E; Milo-Cochavi, Shira; Kerem, Zohar; Eyal, Yoram; Flaishman, Moshe A

    2016-08-01

    Expression of 13 genes encoding chlorophyll biosynthesis and degradation was evaluated. Chlorophyll degradation was differentially regulated in pollinated and parthenocarpic fig fruits, leading to earlier chlorophyll degradation in parthenocarpic fruits. Varieties of the common fig typically yield a commercial summer crop that requires no pollination, although it can be pollinated. Fig fruit pollination results in larger fruit size, greener skin and darker interior inflorescence color, and slows the ripening process compared to non-pollinated fruits. We evaluated the effect of pollination on chlorophyll content and levels of transcripts encoding enzymes of the chlorophyll metabolism in fruits of the common fig 'Brown Turkey'. We cloned and evaluated the expression of 13 different genes. All 13 genes showed high expression in the fruit skin, inflorescences and leaves, but extremely low expression in roots. Pollination delayed chlorophyll breakdown in the ripening fruit skin and inflorescences. This was correlated with the expression of genes encoding enzymes in the chlorophyll biosynthesis and degradation pathways. Expression of pheophorbide a oxygenase (PAO) was strongly negatively correlated with chlorophyll levels during ripening in pollinated fruits; along with its high expression levels in yellow leaves, this supports a pivotal role for PAO in chlorophyll degradation in figs. Normalizing expression levels of all chlorophyll metabolism genes in the pollinated and parthenocarpic fruit skin and inflorescences showed three synthesis (FcGluTR1, FcGluTR2 and FcCLS1) and three degradation (FcCLH1, FcCLH2 and FcRCCR1) genes with different temporal expression in the pollinated vs. parthenocarpic fruit skin and inflorescences. FcCAO also showed different expressions in the parthenocarpic fruit skin. Thus, chlorophyll degradation is differentially regulated in the pollinated and parthenocarpic fruit skin and inflorescences, leading to earlier and more sustained

  8. Factor VIII brand and the incidence of factor VIII inhibitors in previously untreated UK children with severe hemophilia A, 2000-2011

    PubMed Central

    Palmer, Benedict P.; Chalmers, Elizabeth A.; Hart, Daniel P.; Liesner, Ri; Rangarajan, Savita; Talks, Katherine; Williams, Michael; Hay, Charles R. M.

    2014-01-01

    The effect of recombinant factor VIII (rFVIII) brand on inhibitor development was investigated in all 407 severe hemophilia A previously untreated patients born in the United Kingdom (UK) between 1 January 2000 and 31 December 2011. Eighty-eight (22%) had been in the RODIN study. Information was extracted from the National Haemophilia Database. Because exposure days (EDs) were not known for some patients, time from first treatment was used as a surrogate for rFVIII exposure. An inhibitor developed in 118 (29%) patients, 60 high and 58 low titer, after a median (interquartile range) of 7.8 (3.3-13.5) months from first exposure and 16 (9-30) EDs. Of 128 patients treated with Kogenate Bayer/Helixate NexGen, 45 (35.2%, 95% confidence interval [CI] 27.4-43.8) developed an inhibitor compared with 42/172 (24.4%, 95% CI 18.6% to 31.4%) with Advate (P = .04). The adjusted hazard ratio (HR) (95% CI) for Kogenate Bayer/Helixate NexGen compared with Advate was 2.14 (1.12-4.10) (P = .02) for high titer and 1.75 (1.11-2.76) (P = .02) for all inhibitors. When excluding UK-RODIN patients, the adjusted HR (95% CI) for high-titer inhibitors was 2.00 (0.93-4.34) (P = .08). ReFacto AF was associated with a higher incidence of all, but not high-titer, inhibitors than Advate. These results will help inform debate around the relative immunogenicity and use of rFVIII brands. PMID:25339360

  9. Carotenoid biosynthesis changes in five red pepper (Capsicum annuum L.) cultivars during ripening. Cultivar selection for breeding.

    PubMed

    Hornero-Méndez, D; Gómez-Ladrón De Guevara, R; Mínguez-Mosquera, M I

    2000-09-01

    Changes in the biosynthesis of individual carotenoid pigments have been investigated during fruit ripening of five cultivars of red pepper (Capsicum annuum L.): Mana, Numex, Belrubi, Delfin, and Negral (a chlorophyll-retaining mutant when ripe). The study was carried out throughout the ripening process, and with special emphasis on the ripe stage, to discover possible differences between cultivars and to characterize these by their carotenoid pattern and content for selecting the best varieties for breeding programs. Ripening fruit of the five cultivars showed the typical and characteristic pattern of carotenoid biosynthesis for the Capsicum genus. In the five cultivars, lutein and neoxanthin, both characteristic chloroplast pigments, decreased in concentration with ripening and eventually disappeared. beta-Carotene, antheraxanthin, and violaxanthin increased in concentration, and other pigments were biosynthesized de novo: zeaxanthin, beta-cryptoxanthin, capsanthin, capsorubin, capsanthin-5,6-epoxide, and cucurbitaxanthin A. A pool of zeaxanthin stands out of the rest of pigment during ripening, which reveals the importance of this pigment as a branching point in the carotenoid biosynthesis in Capsicum. Quantitatively, Negral cultivar showed the highest increase in total carotenoid content (48. 39-fold), followed by Mana and Delfin with 38.03- and 36.8-fold, respectively, and by Belrubi and Numex with 28.03- and 23.48-fold, respectively. In all the red varieties, there was an inverse relationship between total carotenoid content and the red to yellow isochromic pigment fraction ratio (R/Y) and the capsanthin-to-zeaxanthin ratio (Caps/Zeax). This seems to be related to the carotenogenic capacity of the cultivar, and thus selection and breeding should not only seek a higher total carotenoid content but also attempt to increase these ratios. In the present study, the cultivar Mana had the highest total carotenoid content (13 208 mg/kg dwt), but the lowest R/Y (1

  10. Chromium(iii) oxidation by biogenic manganese oxides with varying structural ripening.

    PubMed

    Tang, Yuanzhi; Webb, Samuel M; Estes, Emily R; Hansel, Colleen M

    2014-09-20

    Manganese (Mn) oxides, which are generally considered biogenic in origin within natural systems, are the only oxidants of Cr(iii) under typical environmental conditions. Yet the influence of Mn biooxide mineral structural evolution on Cr(iii) oxidation under varying geochemical conditions is unknown. In this study we examined the role of light, organic carbon, pH, and the structure of biogenic Mn oxides on Cr(iii) oxidation. Aging of Mn oxides produced by a marine bacterium within the widespread Roseobacter clade resulted in structural ripening from a colloidal hexagonal to a particulate triclinic birnessite phase. The structurally diverse Mn oxides were then reacted with aqueous Cr(iii) within artificial seawater in the presence or absence of carbon and light. Here we found that Cr(iii) oxidation capacity was highest at near neutral pH and in the combined presence of carbon and light. Mn oxide ripening from a hexagonal to a triclinic birnessite phase led to decreased Cr(iii) oxidation in the presence of carbon and light, whereas no change in reactivity was observed in the absence of carbon and/or in the dark. As only minimal Cr(iii) oxidation was observed in the absence of Mn oxides, these results strongly point to coupled Mn oxide- and photo-induced generation of organic and/or oxygen radicals involved in Cr(iii) oxidation. Based on Mn oxide concentration and structural trends, we postulate that Mn(ii) produced from the oxidation of Cr(iii) by the primary Mn oxide is recycled in the presence of organics and light conditions, (re)generating secondary hexagonal birnessite and thereby allowing for continuous oxidation of Cr(iii). In the absence of this Mn oxide regeneration, Cr(iii) induced structural ripening of the hexagonal birnessite precludes further Cr(iii) oxidation. These results highlight the complexity of reactions involved in Mn oxide mediated Cr(iii) oxidation and suggest that photochemical carbon reactions are requisite for sustained Cr(iii) oxidation

  11. Water deficits accelerate ripening and induce changes in gene expression regulating flavonoid biosynthesis in grape berries.

    PubMed

    Castellarin, Simone D; Matthews, Mark A; Di Gaspero, Gabriele; Gambetta, Gregory A

    2007-12-01

    Water deficits consistently promote higher concentrations of anthocyanins in red winegrapes and their wines. However, controversy remains as to whether there is any direct effect on berry metabolism other than inhibition of growth. Early (ED) and late (LD) season water deficits, applied before or after the onset of ripening (veraison), were imposed on field grown Vitis vinifera "Cabernet Sauvignon", and the responses of gene expression in the flavonoid pathway and their corresponding metabolites were determined. ED accelerated sugar accumulation and the onset of anthocyanin synthesis. Both ED and LD increased anthocyanin accumulation after veraison. Expression profiling revealed that the increased anthocyanin accumulation resulted from earlier and greater expression of the genes controlling flux through the anthocyanin biosynthetic pathway, including F3H, DFR, UFGT and GST. Increases in total anthocyanins resulted predominantly from an increase of 3'4'5'-hydroxylated forms through the differential regulation of F3'H and F3'5'H. There were limited effects on proanthocyanidin, other flavonols, and on expression of genes committed to their synthesis. These results demonstrate that manipulation of abiotic stress through applied water deficits not only modulates compositional changes during berry ripening, but also alters the timing of particular aspects of the ripening process.

  12. Neuroprotection with a brain-penetrating biologic tumor necrosis factor inhibitor.

    PubMed

    Zhou, Qing-Hui; Sumbria, Rachita; Hui, Eric Ka-Wai; Lu, Jeff Zhiqiang; Boado, Ruben J; Pardridge, William M

    2011-11-01

    Biologic tumor necrosis factor (TNF)-α inhibitors do not cross the blood-brain barrier (BBB). A BBB-penetrating TNF-α inhibitor was engineered by fusion of the extracellular domain of the type II human TNF receptor (TNFR) to the carboxyl terminus of the heavy chain of a mouse/rat chimeric monoclonal antibody (MAb) against the mouse transferrin receptor (TfR), and this fusion protein is designated cTfRMAb-TNFR. The cTfRMAb-TNFR fusion protein and etanercept bound human TNF-α with high affinity and K(D) values of 374 ± 77 and 280 ± 80 pM, respectively. Neuroprotection in brain in vivo after intravenous administration of the fusion protein was examined in a mouse model of Parkinson's disease. Mice were also treated with saline or a non-BBB-penetrating TNF decoy receptor, etanercept. After intracerebral injection of the nigral-striatal toxin, 6-hydroxydopamine, mice were treated every other day for 3 weeks. Treatment with the cTfRMAb-TNFR fusion protein caused an 83% decrease in apomorphine-induced rotation, a 67% decrease in amphetamine-induced rotation, a 82% increase in vibrissae-elicited forelimb placing, and a 130% increase in striatal tyrosine hydroxylase (TH) enzyme activity. In contrast, chronic treatment with etanercept, which does not cross the BBB, had no effect on neurobehavior or striatal TH enzyme activity. A bridging enzyme-linked immunosorbent assay specific for the cTfRMAb-TNFR fusion protein showed that the immune response generated in the mice was low titer. In conclusion, a biologic TNF inhibitor is neuroprotective after intravenous administration in a mouse model of neurodegeneration, providing that the TNF decoy receptor is reengineered to cross the BBB.

  13. Preclinical Evaluation of MET Inhibitor INC-280 With or Without the Epidermal Growth Factor Receptor Inhibitor Erlotinib in Non–Small-Cell Lung Cancer

    PubMed Central

    Lara, Matthew S.; Holland, William S.; Chinn, Danielle; Burich, Rebekah A.; Lara, Primo N.; Gandara, David R.; Kelly, Karen; Mack, Philip C.

    2018-01-01

    The MET inhibitor INC-280 restored sensitivity to erlotinib and promoted apoptosis in non–small-cell lung cancer models rendered resistant to erlotinib by hepatocyte growth factor. Background Although the epidermal growth factor receptor (EGFR) inhibitor erlotinib is initially effective in non–small-cell lung cancer (NSCLC) patients with tumors harboring activating mutations of EGFR, most subsequently develop acquired resistance. One recognized resistance mechanism occurs through activation of bypass signaling via the hepatocyte growth factor (HGF)-MET pathway. INC-280 is a small molecule kinase inhibitor of MET. We sought to demonstrate the activity of INC-280 on select NSCLC cell lines both as a single agent and in combination with erlotinib using exogenous HGF to simulate MET up-regulation. Methods Four NSCLC cell lines (HCC827, PC9, H1666, and H358) were treated with either single-agent INC-280 or in combination with erlotinib with or without HGF. The activity of the drug treatments was measured by cell viability assays. Immunoblotting was used to monitor expression of EGFR/pEGFR, MET/pMET, GAB1/pGAB1, AKT/pAKT, and ERK/pERK as well as markers of apoptosis (PARP and capase-3 cleavage) in H1666, HCC827, and PC9. Results As a single agent, INC-280 showed minimal cytotoxicity despite potent inhibition of MET kinase activity at concentrations as low as 10 nM. Addition of HGF prevented erlotinib-induced cell death. The addition of INC280 to HGF-mediated erlotinib-resistant models restored erlotinib sensitivity for all cell lines tested, associated with cleavage of both PARP and caspase-3. In these models, INC-280 treatment was sufficient to restore erlotinib-induced inhibition of MET, GAB1, AKT, and ERK in the presence of HGF. Conclusion Although the MET inhibitor INC-280 alone had no discernible effect on cell growth, it was able to restore sensitivity to erlotinib and promote apoptosis in NSCLC models rendered erlotinib resistant by HGF. These data provide a

  14. Grape Ripening Is Regulated by Deficit Irrigation/Elevated Temperatures According to Cluster Position in the Canopy

    PubMed Central

    Zarrouk, Olfa; Brunetti, Cecilia; Egipto, Ricardo; Pinheiro, Carla; Genebra, Tânia; Gori, Antonella; Lopes, Carlos M.; Tattini, Massimiliano; Chaves, M. Manuela

    2016-01-01

    The impact of water deficit on berry quality has been extensively investigated during the last decades. Nonetheless, there is a scarcity of knowledge on the performance of varieties exposed to a combination of high temperatures/water stress during the growing season and under vineyard conditions. The objective of this research was to investigate the effects of two irrigation regimes, sustained deficit irrigation (SDI, 30% ETc) and regulated deficit irrigation (RDI, 15% ETc) and of two cluster positions within the canopy (east- and west-exposed sides) on berry ripening in red Aragonez (Tempranillo) grapevines. The study was undertaken for two successive years in a commercial vineyard in South Portugal, monitoring the following parameters: pre-dawn leaf water potential, berry temperature, sugars, polyphenols, abscisic acid (ABA) and related metabolites. Additionally, expression patterns for different transcripts encoding for enzymes responsible for anthocyanin and ABA biosynthesis (VviUFGT, VvNCED1, VvβG1, VviHyd1, VviHyd2) were analyzed. In both years anthocyanin concentration was lower in RDI at the west side (RDIW- the hottest one) from véraison onwards, suggesting that the most severe water stress conditions exacerbated the negative impact of high temperature on anthocyanin. The down-regulation of VviUFGT expression revealed a repression of the anthocyanin synthesis in berries of RDIW, at early stages of berry ripening. At full-maturation, anthocyanin degradation products were detected, being highest at RDIW. This suggests that the negative impact of water stress and high temperature on anthocyanins results from the repression of biosynthesis at the onset of ripening and from degradation at later stages. On the other hand, berries grown under SDI displayed a higher content in phenolics than those under RDI, pointing out for the attenuation of the negative temperature effects under SDI. Irrigation regime and berry position had small effect on free

  15. Grape Ripening Is Regulated by Deficit Irrigation/Elevated Temperatures According to Cluster Position in the Canopy.

    PubMed

    Zarrouk, Olfa; Brunetti, Cecilia; Egipto, Ricardo; Pinheiro, Carla; Genebra, Tânia; Gori, Antonella; Lopes, Carlos M; Tattini, Massimiliano; Chaves, M Manuela

    2016-01-01

    The impact of water deficit on berry quality has been extensively investigated during the last decades. Nonetheless, there is a scarcity of knowledge on the performance of varieties exposed to a combination of high temperatures/water stress during the growing season and under vineyard conditions. The objective of this research was to investigate the effects of two irrigation regimes, sustained deficit irrigation (SDI, 30% ET c ) and regulated deficit irrigation (RDI, 15% ET c ) and of two cluster positions within the canopy (east- and west-exposed sides) on berry ripening in red Aragonez (Tempranillo) grapevines. The study was undertaken for two successive years in a commercial vineyard in South Portugal, monitoring the following parameters: pre-dawn leaf water potential, berry temperature, sugars, polyphenols, abscisic acid (ABA) and related metabolites. Additionally, expression patterns for different transcripts encoding for enzymes responsible for anthocyanin and ABA biosynthesis ( VviUFGT, VvNCED1, Vv β G1, VviHyd1, VviHyd2 ) were analyzed. In both years anthocyanin concentration was lower in RDI at the west side (RDIW- the hottest one) from véraison onwards, suggesting that the most severe water stress conditions exacerbated the negative impact of high temperature on anthocyanin. The down-regulation of VviUFGT expression revealed a repression of the anthocyanin synthesis in berries of RDIW, at early stages of berry ripening. At full-maturation, anthocyanin degradation products were detected, being highest at RDIW. This suggests that the negative impact of water stress and high temperature on anthocyanins results from the repression of biosynthesis at the onset of ripening and from degradation at later stages. On the other hand, berries grown under SDI displayed a higher content in phenolics than those under RDI, pointing out for the attenuation of the negative temperature effects under SDI. Irrigation regime and berry position had small effect on free

  16. Regulation of fruit ascorbic acid concentrations during ripening in high and low vitamin C tomato cultivars

    PubMed Central

    2012-01-01

    Background To gain insight into the regulation of fruit ascorbic acid (AsA) pool in tomatoes, a combination of metabolite analyses, non-labelled and radiolabelled substrate feeding experiments, enzyme activity measurements and gene expression studies were carried out in fruits of the ‘low-’ and ‘high-AsA’ tomato cultivars ‘Ailsa Craig’ and ‘Santorini’ respectively. Results The two cultivars exhibited different profiles of total AsA (totAsA, AsA + dehydroascorbate) and AsA accumulation during ripening, but both displayed a characteristic peak in concentrations at the breaker stage. Substrate feeding experiments demonstrated that the L-galactose pathway is the main AsA biosynthetic route in tomato fruits, but that substrates from alternative pathways can increase the AsA pool at specific developmental stages. In addition, we show that young fruits display a higher AsA biosynthetic capacity than mature ones, but this does not lead to higher AsA concentrations due to either enhanced rates of AsA breakdown (‘Ailsa Craig’) or decreased rates of AsA recycling (‘Santorini’), depending on the cultivar. In the later stages of ripening, differences in fruit totAsA-AsA concentrations of the two cultivars can be explained by differences in the rate of AsA recycling activities. Analysis of the expression of AsA metabolic genes showed that only the expression of one orthologue of GDP-L-galactose phosphorylase (SlGGP1), and of two monodehydroascorbate reductases (SlMDHAR1 and SlMDHAR3) correlated with the changes in fruit totAsA-AsA concentrations during fruit ripening in ‘Ailsa Craig’, and that only the expression of SlGGP1 was linked to the high AsA concentrations found in red ripe ‘Santorini’ fruits. Conclusions Results indicate that ‘Ailsa Craig’ and ‘Santorini’ use complementary mechanisms to maintain the fruit AsA pool. In the low-AsA cultivar (‘Ailsa Craig’), alternative routes of AsA biosynthesis may supplement biosynthesis via L

  17. Effect of nitrate and nitrite curing salts on microbial changes and sensory quality of rapid ripened sausages.

    PubMed

    Sanz, Y; Vila, R; Toldrá, F; Nieto, P; Flores, J

    1997-07-22

    The effect of the use of either nitrite or nitrate curing salts on microbial changes and sensory quality in rapid ripened sausages inoculated with a mixed starter culture (Lactobacillus sake and Staphylococcus carnosus) was investigated. Lactic acid bacteria and Micrococcaceae were not greatly affected by the added curing salt. Conversely, the inhibition exerted by nitrite on the undesirable flora (Enterobacteriaceae and psychrotrophs) was evident from the early stages of the processing keeping highly significant differences (P < 0.01) with respect to nitrate made sausages till the end of the ripening stage. The use of nitrite in sausage processing was found to reduce hygienic risks.

  18. Metatranscriptome analysis of fungal strains Penicillium camemberti and Geotrichum candidum reveal cheese matrix breakdown and potential development of sensory properties of ripened Camembert-type cheese

    PubMed Central

    2014-01-01

    Background Camembert-type cheese ripening is driven mainly by fungal microflora including Geotrichum candidum and Penicillium camemberti. These species are major contributors to the texture and flavour of typical bloomy rind cheeses. Biochemical studies showed that G. candidum reduces bitterness, enhances sulphur flavors through amino acid catabolism and has an impact on rind texture, firmness and thickness, while P. camemberti is responsible for the white and bloomy aspect of the rind, and produces enzymes involved in proteolysis and lipolysis activities. However, very little is known about the genetic determinants that code for these activities and their expression profile over time during the ripening process. Results The metatranscriptome of an industrial Canadian Camembert-type cheese was studied at seven different sampling days over 77 days of ripening. A database called CamemBank01was generated, containing a total of 1,060,019 sequence tags (reads) assembled in 7916 contigs. Sequence analysis revealed that 57% of the contigs could be affiliated to molds, 16% originated from yeasts, and 27% could not be identified. According to the functional annotation performed, the predominant processes during Camembert ripening include gene expression, energy-, carbohydrate-, organic acid-, lipid- and protein- metabolic processes, cell growth, and response to different stresses. Relative expression data showed that these functions occurred mostly in the first two weeks of the ripening period. Conclusions These data provide further advances in our knowledge about the biological activities of the dominant ripening microflora of Camembert cheese and will help select biological markers to improve cheese quality assessment. PMID:24670012

  19. Metatranscriptome analysis of fungal strains Penicillium camemberti and Geotrichum candidum reveal cheese matrix breakdown and potential development of sensory properties of ripened Camembert-type cheese.

    PubMed

    Lessard, Marie-Hélène; Viel, Catherine; Boyle, Brian; St-Gelais, Daniel; Labrie, Steve

    2014-03-26

    Camembert-type cheese ripening is driven mainly by fungal microflora including Geotrichum candidum and Penicillium camemberti. These species are major contributors to the texture and flavour of typical bloomy rind cheeses. Biochemical studies showed that G. candidum reduces bitterness, enhances sulphur flavors through amino acid catabolism and has an impact on rind texture, firmness and thickness, while P. camemberti is responsible for the white and bloomy aspect of the rind, and produces enzymes involved in proteolysis and lipolysis activities. However, very little is known about the genetic determinants that code for these activities and their expression profile over time during the ripening process. The metatranscriptome of an industrial Canadian Camembert-type cheese was studied at seven different sampling days over 77 days of ripening. A database called CamemBank01 was generated, containing a total of 1,060,019 sequence tags (reads) assembled in 7916 contigs. Sequence analysis revealed that 57% of the contigs could be affiliated to molds, 16% originated from yeasts, and 27% could not be identified. According to the functional annotation performed, the predominant processes during Camembert ripening include gene expression, energy-, carbohydrate-, organic acid-, lipid- and protein- metabolic processes, cell growth, and response to different stresses. Relative expression data showed that these functions occurred mostly in the first two weeks of the ripening period. These data provide further advances in our knowledge about the biological activities of the dominant ripening microflora of Camembert cheese and will help select biological markers to improve cheese quality assessment.

  20. Dynamic changes in proteins during apple (Malus x domestica) fruit ripening and storage

    PubMed Central

    Shi, Yun; Jiang, Li; Zhang, Li; Kang, Ruoyi; Yu, Zhifang

    2014-01-01

    A proteomic study, using two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight, was conducted in apple fruit (cv. ‘Golden Delicious’) starting at 10 days prior to harvest through 50 days in storage. Total protein was extracted using a phenol/sodium dodecyl sulfate protocol. More than 400 protein spots were detected in each gel and 55 differentially expressed proteins (p<0.05) were subjected to matrix-assisted laser desorption/ionization time-of-flight/time-of-flight analysis. Fifty-three of these proteins were finally identified using an apple expressed sequence tag database downloaded from Genome Database for Rosaceae and placed into six categories. The categories and the percentage of proteins placed in each category were stress response and defense (49.0%), energy and metabolism (34.0%), fruit ripening and senescence (5.6%), signal transduction (3.8%), cell structure (3.8%) and protein synthesis (3.8%). Proteins involved in several multiple metabolic pathways, including glycolysis, pentose–phosphate pathway, anti-oxidative systems, photosynthesis and cell wall synthesis, were downregulated, especially during the climacteric burst in respiration and during the senescent stages of fruit development. Proteins classified as allergens or involved in cell wall degradation were upregulated during the ripening process. Some protein spots exhibited a mixed pattern (increasing to maximal abundance followed by a decrease), such as 1-aminocyclopropane-1-carboxylate oxidase, L-ascorbate peroxidase and abscisic acid response proteins. The identification of differentially expressed proteins associated with physiological processes identified in the current study provides a baseline of information for understanding the metabolic processes and regulatory mechanisms that occur in climacteric apple fruit during ripening and senescence. PMID:26504530

  1. Factor VIII inhibitor in a patient with mild haemophilia A and an Asn618-->Ser mutation responsive to immune tolerance induction and cyclophosphamide.

    PubMed

    Vlot, André J; Wittebol, Shulamiet; Strengers, Paul F W; Turenhout, Ellen A M; Voorberg, Jan; van den Berg, H Marijke; Mauser-Bunschoten, Eveline P

    2002-04-01

    We describe a patient with mild haemophilia A (original value of factor VIII activity 0.30 U/ml) who developed an inhibitor (36.1 Bethesda U/ml) which cross-reacted with his endogenous factor VIII. This caused a decline in basal factor VIII level (< 0.01 U/ml) and severe haemorrhagic events. Treatment to induce immune tolerance was started with factor VIII/von Willebrand factor (VWF) concentrates, but inhibitor levels increased progressively and the patient suffered serious bleeding. Cyclophosphamide was administered and, after 8 months treatment, factor VIII levels increased to 0.20 U/ml and the inhibitor could no longer be detected. Screening of his factor VIII gene revealed a missense mutation in exon 13 that predicts substitution of Asn618-->Ser in the A2 domain of factor VIII. Immunoprecipitation analysis showed that the antibodies present in the patient's plasma reacted with metabolically labelled A2 domain and, to a lesser extent, with factor VIII light chain. Inhibitory antibodies were completely neutralized by recombinant A2 domain, whereas no neutralization was observed after the addition of factor VIII light chain (A3-C1-C2) and C2 domain. More detailed analysis showed that the majority of inhibitory antibodies were directed against residues Arg484-Ile508, a previously identified binding site for factor VIII inhibitors. Our findings suggest that immune tolerance therapy and cyclophosphamide were successful in eradicating inhibitory antibodies against a common epitope on factor VIII.

  2. Influence of grape density and harvest date on changes in phenolic composition, phenol extractability indices, and instrumental texture properties during ripening.

    PubMed

    Rolle, Luca; Segade, Susana Río; Torchio, Fabrizio; Giacosa, Simone; Cagnasso, Enzo; Marengo, Fabio; Gerbi, Vincenzo

    2011-08-24

    Changes in the phenolic composition, phenol extractability indices, and mechanical properties occur in grape berries during the ripening process, but the heterogeneity of the grapes harvested at different ripening stages affects the reliability of the results obtained. In this work, these changes were studied in Nebbiolo grapes harvested during five consecutive weeks and then separated according to three density classes. The changes observed in chemical and mechanical parameters through the ripening process are more related to berry density than harvest date. Therefore, the winemaker has to select the flotation density according to the objective quality properties of the wine to be elaborated. On the other hand, the stiffer grapes were associated with a higher accumulation of proanthocyanidins. The harder grapes provided the higher concentration and extractability of flavanols reactive to vanillin, whereas the thicker ones facilitated the extraction of proanthocyanidins.

  3. Discovery of natural mouse serum derived HIV-1 entry inhibitor(s).

    PubMed

    Wei, M; Chen, Y; Xi, J; Ru, S; Ji, M; Zhang, D; Fang, Q; Tang, B

    Among rationally designed human immunodeficiency virus 1 (HIV-1) inhibitors, diverse natural factors have showed as potent anti-HIV activity in human blood. We have discovered that the boiled supernatant of healthy mouse serum could suppress HIV-1 entry, and exhibited reduced inhibitory activity after trypsin digestion. Further analysis demonstrated that only the fraction containing 10-25 K proteins could inhibit HIV-1 mediated cell-cell fusion. These results suggest that the 10-25 K protein(s) is novel natural HIV-1 entry inhibitor(s). Our findings provide important information about novel natural HIV entry inhibitors in mouse serum.

  4. Restoring E-cadherin expression increases sensitivity to epidermal growth factor receptor inhibitors in lung cancer cell lines.

    PubMed

    Witta, Samir E; Gemmill, Robert M; Hirsch, Fred R; Coldren, Christopher D; Hedman, Karla; Ravdel, Larisa; Helfrich, Barbara; Dziadziuszko, Rafal; Chan, Daniel C; Sugita, Michio; Chan, Zeng; Baron, Anna; Franklin, Wilbur; Drabkin, Harry A; Girard, Luc; Gazdar, Adi F; Minna, John D; Bunn, Paul A

    2006-01-15

    The epidermal growth factor receptor (EGFR) is overexpressed in the majority of non-small cell lung cancers (NSCLC). EGFR tyrosine kinase inhibitors, such as gefitinib and erlotinib, produce 9% to 27% response rates in NSCLC patients. E-Cadherin, a calcium-dependent adhesion molecule, plays an important role in NSCLC prognosis and progression, and interacts with EGFR. The zinc finger transcriptional repressor, ZEB1, inhibits E-cadherin expression by recruiting histone deacetylases (HDAC). We identified a significant correlation between sensitivity to gefitinib and expression of E-cadherin, and ZEB1, suggesting their predictive value for responsiveness to EGFR-tyrosine kinase inhibitors. E-Cadherin transfection into a gefitinib-resistant line increased its sensitivity to gefitinib. Pretreating resistant cell lines with the HDAC inhibitor, MS-275, induced E-cadherin along with EGFR and led to a growth-inhibitory and apoptotic effect of gefitinib similar to that in gefitinib-sensitive NSCLC cell lines including those harboring EGFR mutations. Thus, combined HDAC inhibitor and gefitinib treatment represents a novel pharmacologic strategy for overcoming resistance to EGFR inhibitors in patients with lung cancer.

  5. Discovery of Novel Human Epidermal Growth Factor Receptor-2 Inhibitors by Structure-based Virtual Screening.

    PubMed

    Shi, Zheng; Yu, Tian; Sun, Rong; Wang, Shan; Chen, Xiao-Qian; Cheng, Li-Jia; Liu, Rong

    2016-01-01

    Human epidermal growth factor receptor-2 (HER2) is a trans-membrane receptor like protein, and aberrant signaling of HER2 is implicated in many human cancers, such as ovarian cancer, gastric cancer, and prostate cancer, most notably breast cancer. Moreover, it has been in the spotlight in the recent years as a promising new target for therapy of breast cancer. Since virtual screening has become an integral part of the drug discovery process, it is of great significant to identify novel HER2 inhibitors by structure-based virtual screening. In this study, we carried out a series of elegant bioinformatics approaches, such as virtual screening and molecular dynamics (MD) simulations to identify HER2 inhibitors from Food and Drug Administration-approved small molecule drug as potential "new use" drugs. Molecular docking identified top 10 potential drugs which showed spectrum affinity to HER2. Moreover, MD simulations suggested that ZINC08214629 (Nonoxynol-9) and ZINC03830276 (Benzonatate) might exert potential inhibitory effects against HER2-targeted anti-breast cancer therapeutics. Together, our findings may provide successful application of virtual screening studies in the lead discovery process, and suggest that our discovered small molecules could be effective HER2 inhibitor candidates for further study. A series of elegant bioinformatics approaches, including virtual screening and molecular dynamics (MD) simulations were took advantage to identify human epidermal growth factor receptor-2 (HER2) inhibitors. Molecular docking recognized top 10 candidate compounds, which showed spectrum affinity to HER2. Further, MD simulations suggested that ZINC08214629 (Nonoxynol-9) and ZINC03830276 (Benzonatate) in candidate compounds were identified as potential "new use" drugs against HER2-targeted anti-breast cancer therapeutics. Abbreviations used: HER2: Human epidermal growth factor receptor-2, FDA: Food and Drug Administration, PDB: Protein Database Bank, RMSDs: Root mean

  6. A model to assess lactic acid bacteria aminopeptidase activities in Parmigiano Reggiano cheese during ripening.

    PubMed

    Gatti, M; De Dea Lindner, J; Gardini, F; Mucchetti, G; Bevacqua, D; Fornasari, M E; Neviani, E

    2008-11-01

    -specificity amino-peptidase C, glutamyl aminopeptidase A, proline iminopeptidase, and peptidase with high specificity for leucine and alanine. Interestingly, this variable did not affect broad-specificity aminopeptidase N and positively affected X-prolyl dipeptidyl aminopeptidase. The models elaborated varying pH, temperatures, and salt concentration and were a useful, low cost, and fast tool to understand the role of the main peptidases in the different phases of cheese ripening in relation to the major environmental factors influencing enzyme activity.

  7. Insights into the Role of the Berry-Specific Ethylene Responsive Factor VviERF045

    PubMed Central

    Leida, Carmen; Dal Rì, Antonio; Dalla Costa, Lorenza; Gómez, Maria D.; Pompili, Valerio; Sonego, Paolo; Engelen, Kristof; Masuero, Domenico; Ríos, Gabino; Moser, Claudio

    2016-01-01

    During grape ripening, numerous transcriptional and metabolic changes are required in order to obtain colored, sweet, and flavored berries. There is evidence that ethylene, together with other signals, plays an important role in triggering the onset of ripening. Here, we report the functional characterization of a berry-specific Ethylene Responsive Factor (ERF), VviERF045, which is induced just before véraison and peaks at ripening. Phylogenetic analysis revealed it is close to the SHINE clade of ERFs, factors involved in the regulation of wax biosynthesis and cuticle morphology. Transgenic grapevines lines overexpressing VviERF045 were obtained, in vitro propagated, phenotypically characterized, and analyzed for the content of specific classes of metabolites. The effect of VviERF045 was correlated with the level of transgene expression, with high-expressing lines showing stunted growth, discolored and smaller leaves, and a lower level of chlorophylls and carotenoids. One line with intermediate expression, L15, was characterized at the transcriptomic level and showed 573 differentially expressed genes compared to wild type plants. Microscopy and gene expression analyses point toward a major role of VviERF045 in epidermis patterning by acting on waxes and cuticle. They also indicate that VviERF045 affects phenolic secondary metabolism and induces a reaction resembling a plant immune response with modulation of receptor like-kinases and pathogen related genes. These results suggest also a possible role of this transcription factor in berry ripening, likely related to changes in epidermis and cuticle of the berry, cell expansion, a decrease in photosynthetic capacity, and the activation of several defense related genes as well as from the phenylpropanoid metabolism. All these processes occur in the berry during ripening. PMID:28018369

  8. Ethylene and Wound-Induced Gene Expression in the Preclimacteric Phase of Ripening Avocado Fruit and Mesocarp Discs.

    PubMed Central

    Starrett, D. A.; Laties, G. G.

    1993-01-01

    Whereas intact postharvest avocado (Persea americana Mill.) fruit may take 1 or more weeks to ripen, ripening is hastened by pulsing fruit for 24 h with ethylene or propylene and is initiated promptly by cutting slices, or discs, of mesocarp tissue. Because the preclimacteric lag period constitutes the extended and variable component of the ripening syndrome, we postulated that selective gene expression during the lag period leads to the triggering of the climacteric. Accordingly, we sought to identify genes that are expressed gradually in the course of the lag period in intact fruit, are turned on sooner in response to a pulse, and are induced promptly in response to wounding (i.e. slicing). To this end, a mixed cDNA library was constructed from mRNA from untreated fruit, pulsed fruit, and aged slices, and the library was screened for genes induced by wounding or by pulsing and/or wounding. The time course of induction of genes encoding selected clones was established by probing northern blots of mRNA from tissues variously treated over a period of time. Four previously identified ripening-associated genes encoding cellulase, polygalacturonase (PG), cytochrome P-450 oxidase (P-450), and ethylene-forming enzyme (EFE, or 1-aminocyclopropane-1-carboxylic acid synthase), respectively, were studied in the same way. Whereas cellulase, PG, and EFE were ruled out as having a role in the initiation of the climacteric, the time course of P-450 induction, as well as the response of same to pulsing and wounding met the criteria[mdash]together with several clones from the mixed library[mdash]for a gene potentially involved in preclimacteric events leading to the onset of the climacteric. Further, it was established that the continuous presence of ethylene is required for persisting induction, and it is suggested that in selected cases wounding may exert a synergistic effect on ethylene action. PMID:12231929

  9. Variations in zonal fruit starch concentrations of apples – a developmental phenomenon or an indication of ripening?

    PubMed Central

    Doerflinger, Franziska C; Miller, William B; Nock, Jacqueline F; Watkins, Christopher B

    2015-01-01

    Patterns of starch hydrolysis in stem, equatorial, and calyx zones of ‘Honeycrisp’ and ‘Empire’ apples (Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.) during maturation and ripening, and in ‘Gala’ apples in response to propylene or 1-methylcyclopropene (1-MCP) treatments after harvest, were studied. Differences in zonal starch concentrations were found for ‘Empire’ and ‘Gala’ fruits, but not for ‘Honeycrisp’. During maturation and ripening of ‘Empire’, the concentration of starch was highest in the calyx end and lowest in the stem region. Differences in rates of starch hydrolysis among zones were not detected. ‘Honeycrisp’ and ‘Empire’ had the highest concentration of sorbitol in the calyx region, whereas it was highest in the stem-end region in ‘Gala’. The distribution differences of glucose, fructose, and sucrose were similar in all three cultivars; higher fructose and glucose concentrations in the stem region, and higher sucrose concentrations in the calyx end of the fruit. Postharvest treatment of ‘Gala’ with propylene did not affect the internal ethylene concentration of the fruit but 1-MCP markedly inhibited it. Starch concentrations were highest in the calyx end but gradients of starch among zones were not changed by postharvest treatment. The rate of hydrolysis was slowed by 1-MCP treatment, but was unaffected by propylene. Postharvest treatments influenced sorbitol, glucose, and fructose concentrations. Patterns of starch concentration among the zones did not confirm differences in ripening, but reflected its uneven distribution throughout the fruit during development. Therefore, measured differences in zonal starch are most likely related to starch accumulation during fruit development, rather than differences in rates of starch degradation during ripening. PMID:26504584

  10. Dormant and after-Ripened Arabidopsis thaliana Seeds are Distinguished by Early Transcriptional Differences in the Imbibed State

    PubMed Central

    Dekkers, Bas J. W.; Pearce, Simon P.; van Bolderen-Veldkamp, R. P. M.; Holdsworth, Michael J.; Bentsink, Leónie

    2016-01-01

    Seed dormancy is a genetically controlled block preventing the germination of imbibed seeds in favorable conditions. It requires a period of dry storage (after-ripening) or certain environmental conditions to be overcome. Dormancy is an important seed trait, which is under selective pressure, to control the seasonal timing of seed germination. Dormant and non-dormant (after-ripened) seeds are characterized by large sets of differentially expressed genes. However, little information is available concerning the temporal and spatial transcriptional changes during early stages of rehydration in dormant and non-dormant seeds. We employed genome-wide transcriptome analysis on seeds of the model plant Arabidopsis thaliana to investigate transcriptional changes in dry seeds upon rehydration. We analyzed gene expression of dormant and after-ripened seeds of the Cvi accession over four time points and two seed compartments (the embryo and surrounding single cell layer endosperm), during the first 24 h after sowing. This work provides a global view of gene expression changes in dormant and non-dormant seeds with temporal and spatial detail, and these may be visualized via a web accessible tool (http://www.wageningenseedlab.nl/resources). A large proportion of transcripts change similarly in both dormant and non-dormant seeds upon rehydration, however, the first differences in transcript abundances become visible shortly after the initiation of imbibition, indicating that changes induced by after-ripening are detected and responded to rapidly upon rehydration. We identified several gene expression profiles which contribute to differential gene expression between dormant and non-dormant samples. Genes with enhanced expression in the endosperm of dormant seeds were overrepresented for stress-related Gene Ontology categories, suggesting a protective role for the endosperm against biotic and abiotic stress to support persistence of the dormant seed in its environment. PMID

  11. Detection and validation of QTLs for milky-white grains caused by high temperature during the ripening period in Japonica rice

    PubMed Central

    Miyahara, Katsunori; Wada, Takuya; Sonoda, Jun-ya; Tsukaguchi, Tadashi; Miyazaki, Masayuki; Tsubone, Masao; Yamaguchi, Osamu; Ishibashi, Masafumi; Iwasawa, Norio; Umemoto, Takayuki; Kondo, Motohiko

    2017-01-01

    The occurrence of chalky rice (Oryza sativa L.) grains caused by high temperature is a serious problem in rice production. Of the several kinds of chalky grains, milky-white grains are not well analyzed. The milky-white rice grain phenomenon is caused by genetic factors as well as environmental and nutritional conditions. To analyze the genetic control system for rice grain quality, we raised recombinant inbred lines from progeny produced from ‘Tsukushiroman’ (high temperature sensitive) and ‘Chikushi 52’ (high temperature tolerant) cultivars. Quantitative trait locus (QTL) analysis revealed that the QTL on chromosome 4, linked to the simple sequence repeat marker RM16424, contributed substantially to the occurrence of milky-white grains, as it was detected over two experimental years. To validate the effect of the QTL, we developed near isogenic lines that have the ‘Chikushi 52’ segment on the short arm of chromosome 4 in the ‘Tsukushiroman’ genetic background, and that had a lower milky-white grain ratio than that of ‘Tsukushiroman’ when exposed to high temperatures during the ripening period. These results suggest that the ‘Chikushi 52’ allele on chromosome 4 suppresses the occurrence of milky-white grains and improves rice grain quality under heat stress during the grain ripening period. PMID:29085242

  12. Plasmin-dependent proteolysis of tissue factor pathway inhibitor in a mouse model of endotoxemia.

    PubMed

    Lupu, C; Herlea, O; Tang, H; Lijnen, R H; Lupu, F

    2013-01-01

    The development of a procoagulant state in sepsis, owing to aberrant expression of tissue factor (TF) and a sharp decrease in the level of its major inhibitor, TF pathway inhibitor (TFPI), could lead to microthrombotic organ failure. The mechanism for the decline in TFPI activity in the lung could involve plasmin-mediated cleavage of the inhibitor. To investigate the effect of plasmin generation on lung-associated TFPI activity, in normal conditions and during infusion of endotoxin (lipopolysaccharide [LPS]) in mice. Plasmin generation and TFPI activity were assayed in the lungs of mice deficient in tissue-type plasminogen (Plg) activator (t-PA) or Plg, at 2 h after LPS or saline injection. The sharp loss of lung-associated TFPI activity at 2 h after LPS challenge paralleled the abrupt increase in plasmin generation. TFPI activity was significantly retained in both t-PA(-/-) and Plg(-/-) mice, which are unable to generate plasmin. The increased plasmin generation during the early stages of sepsis could cleave/inactivate TFPI and thus lead to thrombotic complications. © 2012 International Society on Thrombosis and Haemostasis.

  13. Regulatory Mechanisms Underlying Oil Palm Fruit Mesocarp Maturation, Ripening, and Functional Specialization in Lipid and Carotenoid Metabolism1[W][OA

    PubMed Central

    Tranbarger, Timothy J.; Dussert, Stéphane; Joët, Thierry; Argout, Xavier; Summo, Marilyne; Champion, Antony; Cros, David; Omore, Alphonse; Nouy, Bruno; Morcillo, Fabienne

    2011-01-01

    Fruit provide essential nutrients and vitamins for the human diet. Not only is the lipid-rich fleshy mesocarp tissue of the oil palm (Elaeis guineensis) fruit the main source of edible oil for the world, but it is also the richest dietary source of provitamin A. This study examines the transcriptional basis of these two outstanding metabolic characters in the oil palm mesocarp. Morphological, cellular, biochemical, and hormonal features defined key phases of mesocarp development. A 454 pyrosequencing-derived transcriptome was then assembled for the developmental phases preceding and during maturation and ripening, when high rates of lipid and carotenoid biosynthesis occur. A total of 2,629 contigs with differential representation revealed coordination of metabolic and regulatory components. Further analysis focused on the fatty acid and triacylglycerol assembly pathways and during carotenogenesis. Notably, a contig similar to the Arabidopsis (Arabidopsis thaliana) seed oil transcription factor WRINKLED1 was identified with a transcript profile coordinated with those of several fatty acid biosynthetic genes and the high rates of lipid accumulation, suggesting some common regulatory features between seeds and fruits. We also focused on transcriptional regulatory networks of the fruit, in particular those related to ethylene transcriptional and GLOBOSA/PISTILLATA-like proteins in the mesocarp and a central role for ethylene-coordinated transcriptional regulation of type VII ethylene response factors during ripening. Our results suggest that divergence has occurred in the regulatory components in this monocot fruit compared with those identified in the dicot tomato (Solanum lycopersicum) fleshy fruit model. PMID:21487046

  14. Null mutation of the MdACS3 gene, coding for a ripening-specific 1-aminocyclopropane-1-carboxylate synthase, leads to long shelf life in apple fruit.

    PubMed

    Wang, Aide; Yamakake, Junko; Kudo, Hisayuki; Wakasa, Yuhya; Hatsuyama, Yoshimichi; Igarashi, Megumi; Kasai, Atsushi; Li, Tianzhong; Harada, Takeo

    2009-09-01

    Expression of MdACS1, coding for 1-aminocyclopropane-1-carboxylate synthase (ACS), parallels the level of ethylene production in ripening apple (Malus domestica) fruit. Here we show that expression of another ripening-specific ACS gene (MdACS3) precedes the initiation of MdACS1 expression by approximately 3 weeks; MdACS3 expression then gradually decreases as MdACS1 expression increases. Because MdACS3 expression continues in ripening fruit treated with 1-methylcyclopropene, its transcription appears to be regulated by a negative feedback mechanism. Three genes in the MdACS3 family (a, b, and c) were isolated from a genomic library, but two of them (MdACS3b and MdACS3c) possess a 333-bp transposon-like insertion in their 5' flanking region that may prevent transcription of these genes during ripening. A single nucleotide polymorphism in the coding region of MdACS3a results in an amino acid substitution (glycine-289 --> valine) in the active site that inactivates the enzyme. Furthermore, another null allele of MdACS3a, Mdacs3a, showing no ability to be transcribed, was found by DNA sequencing. Apple cultivars homozygous or heterozygous for both null allelotypes showed no or very low expression of ripening-related genes and maintained fruit firmness. These results suggest that MdACS3a plays a crucial role in regulation of fruit ripening in apple, and is a possible determinant of ethylene production and shelf life in apple fruit.

  15. Metabolic profiling of a range of peach fruit varieties reveals high metabolic diversity and commonalities and differences during ripening.

    PubMed

    Monti, Laura L; Bustamante, Claudia A; Osorio, Sonia; Gabilondo, Julieta; Borsani, Julia; Lauxmann, Martin A; Maulión, Evangelina; Valentini, Gabriel; Budde, Claudio O; Fernie, Alisdair R; Lara, María V; Drincovich, María F

    2016-01-01

    Peach (Prunus persica) fruits from different varieties display differential organoleptic and nutritional properties, characteristics related to their chemical composition. Here, chemical biodiversity of peach fruits from fifteen varieties, at harvest and after post-harvest ripening, was explored by gas chromatography-mass spectrometry. Metabolic profiling revealed that metabolites involved in organoleptic properties (sugars, organic and amino acids), stress tolerance (raffinose, galactinol, maltitol), and with nutritional properties (amino, caffeoylquinic and dehydroascorbic acids) displayed variety-dependent levels. Peach varieties clustered into four groups: two groups of early-harvest varieties with higher amino acid levels; two groups of mid- and late-harvest varieties with higher maltose levels. Further separation was mostly dependent on organic acids/raffinose levels. Variety-dependent and independent metabolic changes associated with ripening were detected; which contribute to chemical diversity or can be used as ripening markers, respectively. The great variety-dependent diversity in the content of metabolites that define fruit quality reinforces metabolomics usage as a tool to assist fruit quality improvement in peach. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Influence of calcium and phosphorus, lactose, and salt-to-moisture ratio on Cheddar cheese quality: proteolysis during ripening.

    PubMed

    Upreti, P; Metzger, L E; Hayes, K D

    2006-02-01

    Proteolysis in cheese is influenced by the state of proteins (protein-calcium-phosphate interactions), level of indigenous milk enzymes (plasmin), externally added milk-clotting enzymes (chymosin), and endogenous and exogenous enzymes from starter and non-starter lactic acid bacteria (NSLAB). The objective of this study was to determine how different levels of calcium (Ca) and phosphorus (P), residual lactose, and salt-to-moisture ratio (S/M) in cheese influence proteolysis during ripening. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), 2 levels of lactose at pressing (2.4 vs. 0.78%), and 2 levels of S/M (6.4 vs. 4.8%) were manufactured. The cheeses were analyzed for changes in pH 4.6-soluble N, and starter and NSLAB counts during 48 wk of ripening. Cheeses at d 1 were also analyzed for residual chymosin, plasmin, and plasminogen activity. A significant increase in soluble N was observed during ripening for all the treatments. Cheeses with low Ca and P, low lactose, and low S/M treatments exhibited higher levels of proteolysis as compared to their corresponding high treatments. Differences in the rate of proteolysis for cheeses with different levels of Ca and P might be due to changes in protein conformation and differences in residual chymosin in the cheeses. Cheeses with low Ca and P were manufactured by lowering the pH at set and drain, which led to higher chymosin retention in cheeses with low Ca and P compared with high Ca and P. Differences in proteolysis between treatments with different levels of lactose were also partly attributed to residual chymosin activity. In all treatments, a major fraction of plasmin existed as plasminogen, indicating minimal contribution of plasmin to proteolysis in Cheddar cheeses. The number of starter bacteria, in all treatments, decreased significantly during ripening. However, the decrease was larger in the case of high S/M treatments compared with low S/M treatments. In contrast

  17. Restoring conjunctival tolerance by topical nuclear factor-κB inhibitors reduces preservative-facilitated allergic conjunctivitis in mice.

    PubMed

    Guzmán, Mauricio; Sabbione, Florencia; Gabelloni, María Laura; Vanzulli, Silvia; Trevani, Analía Silvina; Giordano, Mirta Nilda; Galletti, Jeremías Gastón

    2014-09-04

    To evaluate the role of nuclear factor-κB (NF-κB) activation in eye drop preservative toxicity and the effect of topical NF-κB inhibitors on preservative-facilitated allergic conjunctivitis. Balb/c mice were instilled ovalbumin (OVA) combined with benzalkonium chloride (BAK) and/or NF-κB inhibitors in both eyes. After immunization, T-cell responses and antigen-induced ocular inflammation were evaluated. Nuclear factor-κB activation and associated inflammatory changes also were assessed in murine eyes and in an epithelial cell line after BAK exposure. Benzalkonium chloride promoted allergic inflammation and leukocyte infiltration of the conjunctiva. Topical NF-κB inhibitors blocked the disruptive effect of BAK on conjunctival immunological tolerance and ameliorated subsequent ocular allergic reactions. In line with these findings, BAK induced NF-κB activation and the secretion of IL-6 and granulocyte-monocyte colony-stimulating factor in an epithelial cell line and in the conjunctiva of instilled mice. In addition, BAK favored major histocompatibility complex (MHC) II expression in cultured epithelial cells in an NF-κB-dependent fashion after interaction with T cells. Benzalkonium chloride triggers conjunctival epithelial NF-κB activation, which seems to mediate some of its immune side effects, such as proinflammatory cytokine release and increased MHC II expression. Breakdown of conjunctival tolerance by BAK favors allergic inflammation, and this effect can be prevented in mice by topical NF-κB inhibitors. These results suggest a new pharmacological target for preservative toxicity and highlight the importance of conjunctival tolerance in ocular surface homeostasis. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

  18. Hydrolytic Enzyme Activities and Protein Pattern of Avocado Fruit Ripened in Air and in Low Oxygen, with and without Ethylene 1

    PubMed Central

    Kanellis, Angelos K.; Solomos, Theophanes; Mattoo, Autar K.

    1989-01-01

    The effect of 2.5% O2 atmosphere with and without ethylene on the activities of hydrolytic enzymes associated with cell walls, and total protein profile during ripening of avocado fruits (Persea americana Mill., cv Hass) were investigated. The low 2.5% O2 atmosphere prevented the rise in the activities of cellulase, polygalacturonase, and acid phosphatase in avocado fruits whose ripening was initiated with ethylene. Addition of 100 microliters per liter ethylene to low O2 atmosphere did not alter these suppressive effects of 2.5% O2. Furthermore, 2.5% O2 atmosphere delayed the development of a number of polypeptides that appear during ripening of avocado fruits while at the same time new polypeptides accumulated. The composition of the extraction buffer and its pH greatly affected the recovery of cellulase activity and its total immunoreactive protein. Images Figure 1 Figure 2 Figure 5 PMID:16666746

  19. Physicochemical and Sensory Properties of Appenzeller Cheese Supplemented with Powdered Microcapsule of Tomato Extract during Ripening

    PubMed Central

    Kwak, Hae-Soo; Chimed, Chogsom; Yoo, Sang-Hun

    2016-01-01

    The objective of this study was to determine the physicochemical and sensory properties of Appenzeller cheese supplemented with different concentrations (0, 1, 2, 3, and 4%, w/w) of powdered microcapsules of tomato extracts (PMT) during ripening at 14℃ for 6 mon. The particle sizes of PMT ranged from 1 to 10 m diameter with an average particle size of approximately 2 m. Butyric acid (C4) concentrations of PMT-added Appenzeller cheese were significantly higher than that of the control. Lactic acid bacteria counts in the cheese were not significantly influenced by ripening time from 0 to 6 mon or the concentrations (0-4%, w/w) of PMT. In terms of texture, the hardness of PMT-added Appenzeller cheese was significantly increased compared to the control. The gumminess and chewiness of PMT-added Appenzeller cheese were similar to those of the control. However, both cohesiveness and springiness of PMT-added Appenzeller cheese were slightly decreased. In sensory analysis, bitterness and sourness of Appenzeller cheese were not significantly changed after supplementation of PMT, but sweetness of the cheese was significantly increased after increasing the ripening time from 0 to 6 mon and increasing the concentration from 1 to 4% (w/w). Based on these results, the addition of the concentrations (1-4%, w/w) of PMT to Appenzeller cheese can be used to develop functional Appenzeller cheese. PMID:27194934

  20. Evaluation of bacterial flora during the ripening of Kedong sufu, a typical Chinese traditional bacteria-fermented soybean product.

    PubMed

    Feng, Zhen; Gao, Wei; Ren, Dan; Chen, Xi; Li, Juan-juan

    2013-04-01

    Kedong sufu is a typical bacteria-fermented sufu in China. Isolation and identification of the autochthonous bacteria involved would allow the design of specific starters for this speciality. The purpose of the present study was to evaluate the bacterial flora during the ripening of Kedong sufu using polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) and culturing. In terms of bacterial diversity, 22 strains were isolated and identified and 27 strains were detected by DGGE. Regarding bacterial dynamics, the results of culturing and PCR-DGGE exhibited a similar trend towards dominant strains. Throughout the fermentation of sufu, Enterococcus avium, Enterococcus faecalis and Staphylococcus carnosus were the dominant microflora, while the secondary microflora comprised Leuconostoc mesenteroides, Staphylococcus saprophyticus, Streptococcus lutetiensis, Kocuria rosea, Kocuria kristinae, Bacillus pumilus, Bacillus cereus and Bacillus subtilis. This study is the first to reveal the bacterial flora during the ripening of Kedong sufu using both culture-dependent and culture-independent methods. This information will help in the design of autochthonous starter cultures for the production of Kedong sufu with desirable characteristic sensory profiles and shorter ripening times. © 2012 Society of Chemical Industry.

  1. Effect of kafirin protein coating on sensory quality and shelf-life of 'Packham's Triumph' pears during ripening.

    PubMed

    Buchner, Sonya; Kinnear, Marise; Crouch, Ian J; Taylor, Janet; Minnaar, Amanda

    2011-12-01

    Pears are exported in large quantities from South Africa, resulting in large revenues. Minimisation of quality losses once the fruit has reached the export destination is as important as following strict export and distribution protocols. Kafirin can form edible films. In this study an edible coating made from 20 g kg(-1) kafirin coating solution was applied as a postharvest treatment to retard quality deterioration of 'Packham's Triumph' pears during storage at the typical ripening temperature (20 °C). Changes in physicochemical and sensory quality were monitored over a period of 24 days. The kafirin coating was unable to retard the onset of ripening but decreased the respiration rate and retarded the progression of senescence. However, moisture loss was exacerbated in the kafirin-coated fruit during ripening at 20 °C, especially towards the end of the shelf-life. The kafirin coating extended the eat-ripe quality of the pears by 1-2 weeks. However, the appearance of the fruit was unacceptable after 14 days of storage in terms of wrinkled skin. Further work is needed to improve the water barrier properties of the kafirin coating by incorporating a wax or triglyceride into the coating formulation or more simply by applying a kafirin coating to waxed fruit. Copyright © 2011 Society of Chemical Industry.

  2. Ostwald ripening of faceted Si particles in an Al-Si-Cu melt

    DOE PAGES

    Shahani, A. J.; Xiao, X.; Skinner, K.; ...

    2016-07-04

    The microstructural evolution of an Al-Si-Cu alloy during Ostwald ripening is imaged via synchrotron-based, four-dimensional (i.e., space and time resolved) X-ray tomography. Samples of composition Al-32 wt%Si-15 wt%Cu were annealed isothermally at 650 °C, in the two-phase solid-liquid regime, while tomographic projections were collected in situ over the course of five hours. Advances in experimental methods and computational approaches enable us to characterize the local interfacial curvatures and velocities during ripening. The sequence of three-dimensional reconstructions and interfacial shape distributions shows highly faceted Si particles in a copper-enriched liquid, that become increasingly isotropic or rounded over time. In addition, wemore » find that the coarsening rate constant is approximately the same in the binary and ternary systems. By coupling these experimental measurements with CALPHAD modeling and ab initio molecular dynamics simulation, we assess the influence of Cu on the coarsening process. Lastly, we find the unusual “pinning” of microstructure at the junction between rough and smooth interfaces and suggest a mechanism for this behavior.« less

  3. Germination parameterization and development of an after-ripening thermal-time model for primary dormancy release of Lithospermum arvense seeds.

    PubMed

    Chantre, Guillermo R; Batlla, Diego; Sabbatini, Mario R; Orioli, Gustavo

    2009-06-01

    Models based on thermal-time approaches have been a useful tool for characterizing and predicting seed germination and dormancy release in relation to time and temperature. The aims of the present work were to evaluate the relative accuracy of different thermal-time approaches for the description of germination in Lithospermum arvense and to develop an after-ripening thermal-time model for predicting seed dormancy release. Seeds were dry-stored at constant temperatures of 5, 15 or 24 degrees C for up to 210 d. After different storage periods, batches of 50 seeds were incubated at eight constant temperature regimes of 5, 8, 10, 13, 15, 17, 20 or 25 degrees C. Experimentally obtained cumulative-germination curves were analysed using a non-linear regression procedure to obtain optimal population thermal parameters for L. arvense. Changes in these parameters were described as a function of after-ripening thermal-time and storage temperature. The most accurate approach for simulating the thermal-germination response of L. arvense was achieved by assuming a normal distribution of both base and maximum germination temperatures. The results contradict the widely accepted assumption of a single T(b) value for the entire seed population. The after-ripening process was characterized by a progressive increase in the mean maximum germination temperature and a reduction in the thermal-time requirements for germination at sub-optimal temperatures. The after-ripening thermal-time model developed here gave an acceptable description of the observed field emergence patterns, thus indicating its usefulness as a predictive tool to enhance weed management tactics.

  4. The regulation of MADS-box gene expression during ripening of banana and their regulatory interaction with ethylene

    PubMed Central

    Elitzur, Tomer; Vrebalov, Julia; Giovannoni, James J.; Goldschmidt, Eliezer E.; Friedman, Haya

    2010-01-01

    Six MaMADS-box genes have been cloned from the banana fruit cultivar Grand Nain. The similarity of these genes to tomato LeRIN is low and neither MaMADS2 nor MaMADS1 complement the tomato rin mutation. Nevertheless, the expression patterns, specifically in fruit and the induction during ripening and in response to ethylene and 1-MCP, suggest that some of these genes may participate in ripening. MaMADS1, 2, and 3, are highly expressed in fruit only, while the others are expressed in fruit as well as in other organs. Moreover, the suites of MaMADS-box genes and their temporal expression differ in peel and pulp during ripening. In the pulp, the increase in MaMADS2, 3, 4, and 5 expression preceded an increase in ethylene production, but coincides with the CO2 peak. However, MaMADS1 expression in pulp coincided with ethylene production, but a massive increase in its expression occurred late during ripening, together with a second wave in the expression of MaMADS2, 3, and 4. In the peel, on the other hand, an increase in expression of MaMADS1, 3, and to a lesser degree also of MaMADS4 and 2 coincided with an increase in ethylene production. Except MaMADS3, which was induced by ethylene in pulp and peel, only MaMADS4, and 5 in pulp and MaMADS1 in peel were induced by ethylene. 1-MCP applied at the onset of the increase in ethylene production, increased the levels of MaMADS4 and MaMADS1 in pulp, while it decreased MaMADS1, 3, 4, and 5 in peel, suggesting that MaMADS4 and MaMADS1 are negatively controlled by ethylene at the onset of ethylene production only in pulp. Only MaMADS2 is neither induced by ethylene nor by 1-MCP, and it is expressed mainly in pulp. Our results suggest that two independent ripening programs are employed in pulp and peel which involve the activation of mainly MaMADS2, 4, and 5 and later on also MaMADS1 in pulp, and mainly MaMADS1, and 3 in peel. Hence, our results are consistent with MaMADS2, a SEP3 homologue, acting in the pulp upstream of the

  5. The regulation of MADS-box gene expression during ripening of banana and their regulatory interaction with ethylene.

    PubMed

    Elitzur, Tomer; Vrebalov, Julia; Giovannoni, James J; Goldschmidt, Eliezer E; Friedman, Haya

    2010-03-01

    Six MaMADS-box genes have been cloned from the banana fruit cultivar Grand Nain. The similarity of these genes to tomato LeRIN is low and neither MaMADS2 nor MaMADS1 complement the tomato rin mutation. Nevertheless, the expression patterns, specifically in fruit and the induction during ripening and in response to ethylene and 1-MCP, suggest that some of these genes may participate in ripening. MaMADS1, 2, and 3, are highly expressed in fruit only, while the others are expressed in fruit as well as in other organs. Moreover, the suites of MaMADS-box genes and their temporal expression differ in peel and pulp during ripening. In the pulp, the increase in MaMADS2, 3, 4, and 5 expression preceded an increase in ethylene production, but coincides with the CO(2) peak. However, MaMADS1 expression in pulp coincided with ethylene production, but a massive increase in its expression occurred late during ripening, together with a second wave in the expression of MaMADS2, 3, and 4. In the peel, on the other hand, an increase in expression of MaMADS1, 3, and to a lesser degree also of MaMADS4 and 2 coincided with an increase in ethylene production. Except MaMADS3, which was induced by ethylene in pulp and peel, only MaMADS4, and 5 in pulp and MaMADS1 in peel were induced by ethylene. 1-MCP applied at the onset of the increase in ethylene production, increased the levels of MaMADS4 and MaMADS1 in pulp, while it decreased MaMADS1, 3, 4, and 5 in peel, suggesting that MaMADS4 and MaMADS1 are negatively controlled by ethylene at the onset of ethylene production only in pulp. Only MaMADS2 is neither induced by ethylene nor by 1-MCP, and it is expressed mainly in pulp. Our results suggest that two independent ripening programs are employed in pulp and peel which involve the activation of mainly MaMADS2, 4, and 5 and later on also MaMADS1 in pulp, and mainly MaMADS1, and 3 in peel. Hence, our results are consistent with MaMADS2, a SEP3 homologue, acting in the pulp upstream of the

  6. Polyamine metabolism in ripening tomato fruit. II. Polyamine metabolism and synthesis in relation to enhanced putrescine content and storage life of alc tomato fruit

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rastogi, R.; Davies, P.J.

    1991-01-01

    The fruit of the Alcobaca landrace of tomato (Lycopersicon esculentum Mill.) have prolonged keeping qualities (determined by the allele alc) and contain three times as much putrescine as the standard Rutgers variety (Alc) at the ripe stage. Polyamine metabolism and biosynthesis were compared in fruit from Rutgers and Rutgers-alc-a near isogenic line possessing the allele alc, at four different stages of ripening. The levels of soluble polyamine conjugates as well as wall bound polyamines in the pericarp tissue and jelly were very low or nondetectable in both genotypes. The increase in putrescine content in alc pericarp is not related tomore » normal ripening as it occurred with time and whether or not the fruit ripened. Pericarp discs of both normal and alc fruit showed a decrease in the metabolism of (1,4-{sup 14}C)putrescine and (terminal labeled-{sup 3}H)spermidine with ripening, but there were no significant differences between the two genotypes. The activity of ornithine decarboxylase was similar in the fruit pericarp of the two lines. Arginine decarboxylase activity decreased during ripening in Rutgers but decreased and rose again in Rutgers-alc fruit, and as a result it was significantly higher in alc fruit than in the normal fruit at the ripe stage. The elevated putrescine levels in alc fruit appear, therefore, to be due to an increase in the activity of arginine decarboxylase.« less

  7. Recommendations for the Prophylactic Management of Skin Reactions Induced by Epidermal Growth Factor Receptor Inhibitors in Patients With Solid Tumors

    PubMed Central

    Deplanque, Gaël; Komatsu, Yoshito; Kobayashi, Yoshimitsu; Ocvirk, Janja; Racca, Patrizia; Guenther, Silke; Zhang, Jun; Lacouture, Mario E.; Jatoi, Aminah

    2016-01-01

    Inhibition of the epidermal growth factor receptor (EGFR) is an established treatment that extends patient survival across a variety of tumor types. EGFR inhibitors fall into two main categories: anti-EGFR monoclonal antibodies, such as cetuximab and panitumumab, and first-generation tyrosine kinase inhibitors, such as afatinib, gefitinib, and erlotinib. Skin reactions are the most common EGFR inhibitor-attributable adverse event, resulting in papulopustular (acneiform) eruptions that can be painful and debilitating, and which may potentially have a negative impact on patients’ quality of life and social functioning, as well as a negative impact on treatment duration. Shortened treatment duration can, in turn, compromise antineoplastic efficacy. Similarly, appropriate management of skin reactions is dependent on their accurate grading; however, conventional means for grading skin reactions are inadequate, particularly within the context of clinical trials. Treating a skin reaction only once it occurs (reactive treatment strategies) may not be the most effective management approach; instead, prophylactic approaches may be preferable. Indeed, we support the viewpoint that prophylactic management of skin reactions should be recommended for all patients treated with EGFR inhibitors. Appropriate prophylactic management could effectively reduce the severity of skin reactions in patients treated with EGFR inhibitors and therefore has the potential to directly benefit patients and improve drug adherence. Accordingly, here we review published and still-emerging data, and provide practical and evidence-based recommendations and algorithms regarding the optimal prophylactic management of EGFR inhibitor-attributable skin reactions. Implications for Practice: Epidermal growth factor receptor (EGFR) inhibitors extend patient survival across a variety of tumor types. The most common EGFR inhibitor-attributable adverse events are skin reactions. Prophylactic—rather than

  8. KRAS Testing and Epidermal Growth Factor Receptor Inhibitor Treatment for Colorectal Cancer in Community Settings

    PubMed Central

    Webster, Jennifer; Kauffman, Tia L.; Feigelson, Heather Spencer; Pawloski, Pamala A.; Onitilo, Adedayo A.; Potosky, Arnold L.; Cross, Deanna; Meier, Paul R.; Mirabedi, Anousheh S.; Delate, Thomas; Daida, Yihe; Williams, Andrew E.; Alexander, Gwen L.; McCarty, Catherine A.; Honda, Stacey; Kushi, Lawrence H.; Goddard, Katrina A.B.

    2013-01-01

    Background In metastatic colorectal cancer (mCRC), mutations in the KRAS gene predict poor response to epidermal growth factor receptor (EGFR) inhibitors. Clinical treatment guidelines now recommend KRAS testing if EGFR inhibitors are considered. Our study investigates the clinical uptake and utilization of KRAS testing. Methods We included 1,188 patients with mCRC diagnosed from 2004 to 2009, from seven integrated health care delivery systems with a combined membership of 5.5 million. We used electronic medical records and targeted manual chart review to capture the complexity and breadth of real-world clinical oncology care. Results Overall, 428 patients (36%) received KRAS testing during their clinical care, and 266 (22%) were treated with EGFR inhibitors. Age at diagnosis (p=0.0034), comorbid conditions (p=0.0316), and survival time from diagnosis (p<0.0001) influence KRAS testing and EGFR inhibitor prescribing. The proportion who received KRAS testing increased from 7% to 97% for those treated in 2006 and 2010, respectively, and 83% of all treated patients had a KRAS wild type genotype. Most patients with a KRAS mutation (86%) were not treated with EGFR inhibitors. The interval between mCRC diagnosis and receipt of KRAS testing decreased from 26 months (2006) to 10 months (2009). Conclusions These findings demonstrate rapid uptake and incorporation of this predictive biomarker into clinical oncology care. Impact In this delivery setting, KRAS testing is widely used to guide treatment decisions with EGFR inhibitors in patients with mCRC. An important future research goal is to evaluate utilization of KRAS testing in other delivery settings in the US. PMID:23155138

  9. Phosphoglucose isomerase from bananas: partial characterization and relation to main changes in carbohydrate composition during ripening.

    PubMed

    Cordenunsi, B R; Oliveira do Nascimento, J R; Vieira da Mota, R; Lajolo, F M

    2001-10-01

    Some characteristics of phosphoglucose isomerase (PGI, EC 5.3.1.9) from banana were measured during fruit ripening of three banana cultivars. In banana, PGI was present as two dimeric isoenzymes, named PGI1 and PGI2, which had similar native molecular masses but differed in relation to heat stability and isoelectric point. Total PGI activity showed a distinct two-step change during fruit ripening. Before the climacteric period, PGI activity gradually decreased with the starch content, then its activity began to increase with sucrose accumulation. The ratio of PGI1, and PGI2 was constant, indicating that both enzymes would be involved in starch degradation and sucrose synthesis. PGI activity and changes in carbohydrate composition suggests the existence of some control to fit the requirements of the intense carbon flow from starch to sucrose.

  10. Gas exchange in fruits related to skin condition and fruit ripening studied with diode laser spectroscopy

    NASA Astrophysics Data System (ADS)

    Huang, Jing; Zhang, Hao; Lin, Huiying; Li, Tianqi; Mei, Liang; Svanberg, Katarina; Svanberg, Sune

    2016-12-01

    The concentration of the biologically active molecular oxygen gas is of crucial importance for fruits in the metabolic respiration, maturation, and ripening processes. In our study, oxygen content and oxygen transport in fruits, exemplified by apples and guavas, were studied noninvasively by gas in scattering media absorption spectroscopy. The technique is based on the fact that free gases typically have 10,000 times narrower absorption features than the bulk material. The technique was demonstrated in studies of the influence of the fruit skin in regulating the internal oxygen balance, by observing the signal response of the internal oxygen gas to a transient change in the ambient gas concentration on peeled and unpeeled fruits. In addition, the gas exchange rate at different ripening stages was also studied in intact guavas.

  11. Gas exchange in fruits related to skin condition and fruit ripening studied with diode laser spectroscopy.

    PubMed

    Huang, Jing; Zhang, Hao; Lin, Huiying; Li, Tianqi; Mei, Liang; Svanberg, Katarina; Svanberg, Sune

    2016-12-01

    The concentration of the biologically active molecular oxygen gas is of crucial importance for fruits in the metabolic respiration, maturation, and ripening processes. In our study, oxygen content and oxygen transport in fruits, exemplified by apples and guavas, were studied noninvasively by gas in scattering media absorption spectroscopy. The technique is based on the fact that free gases typically have 10,000 times narrower absorption features than the bulk material. The technique was demonstrated in studies of the influence of the fruit skin in regulating the internal oxygen balance, by observing the signal response of the internal oxygen gas to a transient change in the ambient gas concentration on peeled and unpeeled fruits. In addition, the gas exchange rate at different ripening stages was also studied in intact guavas.

  12. Epidermal growth factor receptor tyrosine kinase inhibitors: application in non-small cell lung cancer.

    PubMed

    Thomas, Melodie

    2003-12-01

    Despite treatment advances over the past decade, long-term survival for patients with non-small cell lung cancer (NSCLC) remains poor, and treatment options available after second-line therapy are limited. Increased understanding of cancer biology has led to the identification of several potential targets for treatment. The epidermal growth factor receptor (EGFR) belongs to a family of plasma membrane receptor tyrosine kinases that controls many important cellular functions, from growth and proliferation to cell death. This receptor is a particularly promising therapeutic target because it often is overexpressed in patients with NSCLC and has been implicated in the pathogenesis as well as the proliferation, invasion, and metastasis of lung cancer and other malignancies. New agents developed to inhibit EGFR function include small-molecule tyrosine kinase inhibitors, monoclonal antibodies to EGFR, and pan-EGFR inhibitors. Completed and ongoing clinical trials have shown that EGFR inhibitors have remarkable efficacy for patients with relapsed NSCLC. Among these, two phase 2 trials have shown that ZD1839 is effective when used as monotherapy. The response rates are comparable with those for docetaxel given in the second-line setting. Another phase 2 trial has shown that OSI-774 is effective in the same setting. Data from phase 3 trials indicate that adding an EGFR tyrosine kinase inhibitor to chemotherapy does not provide an additional survival benefit, as compared with standard chemotherapy alone for first-line treatment of NSCLC. It appears that EGFR tyrosine kinase inhibitors are safe and well tolerated by patients with cancer. Further studies will elucidate how these new agents can best be used for NSCLC and other tumor types.

  13. XTHs from Fragaria vesca: genomic structure and transcriptomic analysis in ripening fruit and other tissues.

    PubMed

    Opazo, María Cecilia; Lizana, Rodrigo; Stappung, Yazmina; Davis, Thomas M; Herrera, Raúl; Moya-León, María Alejandra

    2017-11-07

    Fragaria vesca or 'woodland strawberry' has emerged as an attractive model for the study of ripening of non-climacteric fruit. It has several advantages, such as its small genome and its diploidy. The recent availability of the complete sequence of its genome opens the possibility for further analysis and its use as a reference species. Fruit softening is a physiological event and involves many biochemical changes that take place at the final stages of fruit development; among them, the remodeling of cell walls by the action of a set of enzymes. Xyloglucan endotransglycosylase/hydrolase (XTH) is a cell wall-associated enzyme, which is encoded by a multigene family. Its action modifies the structure of xyloglucans, a diverse group of polysaccharides that crosslink with cellulose microfibrills, affecting therefore the functional structure of the cell wall. The aim of this work is to identify the XTH-encoding genes present in F. vesca and to determine its transcription level in ripening fruit. The search resulted in identification of 26 XTH-encoding genes named as FvXTHs. Genetic structure and phylogenetic analyses were performed allowing the classification of FvXTH genes into three phylogenetic groups: 17 in group I/II, 2 in group IIIA and 4 in group IIIB. Two sequences were included into the ancestral group. Through a comparative analysis, characteristic structural protein domains were found in FvXTH protein sequences. In complement, expression analyses of FvXTHs by qPCR were performed in fruit at different developmental and ripening stages, as well as, in other tissues. The results showed a diverse expression pattern of FvXTHs in several tissues, although most of them are highly expressed in roots. Their expression patterns are not related to their respective phylogenetic groups. In addition, most FvXTHs are expressed in ripe fruit, and interestingly, some of them (FvXTH 18 and 20, belonging to phylogenic group I/II, and FvXTH 25 and 26 to group IIIB) display an

  14. Severe Extraarticular Manifestations in a Community-based Cohort of Patients with Rheumatoid Arthritis: Risk Factors and Incidence in Relation to Treatment with Tumor Necrosis Factor Inhibitors.

    PubMed

    Theander, Lisa; Nyhäll-Wåhlin, Britt-Marie; Nilsson, Jan-Åke; Willim, Minna; Jacobsson, Lennart T H; Petersson, Ingemar F; Turesson, Carl

    2017-07-01

    The aims of this study were to evaluate whether treatment with tumor necrosis factor (TNF) inhibitors in patients with rheumatoid arthritis (RA) affects the risk of developing severe extraarticular rheumatoid arthritis (ExRA) manifestations and to investigate potential predictors for developing ExRA. A dynamic community-based cohort of patients with RA was studied (n = 1977). Clinical records were reviewed and cases of severe ExRA were identified. Information on exposure to TNF inhibitors was obtained from a regional register. Exposure to TNF inhibitors was analyzed in a time-dependent fashion and the incidence of severe ExRA in exposed patients was compared with the incidence in unexposed patients. Cox regression models were used to assess potential predictors of severe ExRA. During treatment with TNF inhibitors, there were 17 patients with new onset of severe ExRA in 2400 person-years at risk (PY; 0.71/100 PY, 95% CI 0.41-1.13) compared with 104 in 15,599 PY (0.67/100 PY, 95% CI 0.54-0.81) in patients without TNF inhibitors. This corresponded to an incidence rate ratio of 1.06 (95% CI 0.60-1.78). The age- and sex-adjusted HR for ExRA in anti-TNF-treated patients was 1.21 (95% CI 1.02-1.43), with similar findings in models adjusted for time-dependent Health Assessment Questionnaire and propensity for anti-TNF treatment. Male sex, positive rheumatoid factor (RF), long disease duration, and greater disability were predictors for ExRA. This study suggests that patients treated with TNF inhibitors are at a slightly increased risk of developing severe ExRA. RF-positive patients with disabling disease of long duration were more likely to develop severe ExRA.

  15. Exploring a volatomic-based strategy for a fingerprinting approach of Vaccinium padifolium L. berries at different ripening stages.

    PubMed

    Porto-Figueira, Priscilla; Figueira, José A; Berenguer, Pedro; Câmara, José S

    2018-04-15

    The effect of ripening on the evolution of the volatomic pattern from endemic Vaccinium padifolium L. (Uveira) berries was investigated using headspace-solid phase microextraction (HS-SPME) followed by gas chromatography/quadrupole-mass spectrometry (GC-qMS) and multivariate statistical analysis (MVA). The most significant HS-SPME parameters, namely fibre polymer, ionic strength and extraction time, were optimized in order to improve extraction efficiency. Under optimal experimental conditions (DVB/CAR/PDMS fibre coating, 40°C, 30min extraction time and 5g of sample amount), a total of 72 volatiles of different functionalities were isolated and identified. Terpenes followed by higher alcohols and esters were the predominant classes in the ripening stages - green, break and ripe. Although significant differences in the volatomic profiles at the three stages were obtained, cis-β-ocimene (2.0-40.0%), trans-2-hexenol (2.4-19.4%), cis-3-hexenol (2.5.16.4%), β-myrcene (1.9-13.8%), 1-hexanol (1.7-13.6%), 2-hexenal (0.7-8.0%), 2-heptanone (0.7-7.7%), and linalool (1.9-6.1%) were the main volatile compounds identified. Higher alcohols, carboxylic acids and ketones gradually increased during ripening, whereas monoterpenes significantly decreased. These trends were dominated by the higher alcohols (1-hexanol, cis-3-hexenol, trans-2-hexenol) and monoterpenes (β-myrcene, cis-β-ocimene and trans-β-ocimene). Partial least squares regression (PLSR) revealed that ethyl caprylate (1.000), trans-geraniol (0.995), ethyl isovalerate (-0.994) and benzyl carbinol (0.993) are the key variables that most contributed to the successful differentiation of Uveira berries according to ripening stage. To the best of our knowledge, no study has carried out on the volatomic composition of berries from endemic Uveira. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Crystal Structure of a Two-domain Fragment of Hepatocyte Growth Factor Activator Inhibitor-1: FUNCTIONAL INTERACTIONS BETWEEN THE KUNITZ-TYPE INHIBITOR DOMAIN-1 AND THE NEIGHBORING POLYCYSTIC KIDNEY DISEASE-LIKE DOMAIN.

    PubMed

    Hong, Zebin; De Meulemeester, Laura; Jacobi, Annemarie; Pedersen, Jan Skov; Morth, J Preben; Andreasen, Peter A; Jensen, Jan K

    2016-07-01

    Hepatocyte growth factor activator inhibitor-1 (HAI-1) is a type I transmembrane protein and inhibitor of several serine proteases, including hepatocyte growth factor activator and matriptase. The protein is essential for development as knock-out mice die in utero due to placental defects caused by misregulated extracellular proteolysis. HAI-1 contains two Kunitz-type inhibitor domains (Kunitz), which are generally thought of as a functionally self-contained protease inhibitor unit. This is not the case for HAI-1, where our results reveal how interdomain interactions have evolved to stimulate the inhibitory activity of an integrated Kunitz. Here we present an x-ray crystal structure of an HAI-1 fragment covering the internal domain and Kunitz-1. The structure reveals not only that the previously uncharacterized internal domain is a member of the polycystic kidney disease domain family but also how the two domains engage in interdomain interactions. Supported by solution small angle x-ray scattering and a combination of site-directed mutagenesis and functional assays, we show that interdomain interactions not only stabilize the fold of the internal domain but also stimulate the inhibitory activity of Kunitz-1. By completing our structural characterization of the previously unknown N-terminal region of HAI-1, we provide new insight into the interplay between tertiary structure and the inhibitory activity of a multidomain protease inhibitor. We propose a previously unseen mechanism by which the association of an auxiliary domain stimulates the inhibitory activity of a Kunitz-type inhibitor (i.e. the first structure of an intramolecular interaction between a Kunitz and another domain). © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Withaferin A, a natural compound with anti-tumor activity, is a potent inhibitor of transcription factor C/EBPβ.

    PubMed

    Falkenberg, Kim D; Jakobs, Anke; Matern, Julian C; Dörner, Wolfgang; Uttarkar, Sagar; Trentmann, Amke; Steinmann, Simone; Coulibaly, Anna; Schomburg, Caroline; Mootz, Henning D; Schmidt, Thomas J; Klempnauer, Karl-Heinz

    2017-07-01

    Recent work has shown that deregulation of the transcription factor Myb contributes to the development of leukemia and several other human cancers, making Myb and its cooperation partners attractive targets for drug development. By employing a myeloid Myb-reporter cell line we have identified Withaferin A (WFA), a natural compound that exhibits anti-tumor activities, as an inhibitor of Myb-dependent transcription. Analysis of the inhibitory mechanism of WFA showed that WFA is a significantly more potent inhibitor of C/EBPβ, a transcription factor cooperating with Myb in myeloid cells, than of Myb itself. We show that WFA covalently modifies specific cysteine residues of C/EBPβ, resulting in the disruption of the interaction of C/EBPβ with the co-activator p300. Our work identifies C/EBPβ as a novel direct target of WFA and highlights the role of p300 as a crucial co-activator of C/EBPβ. The finding that WFA is a potent inhibitor of C/EBPβ suggests that inhibition of C/EBPβ might contribute to the biological activities of WFA. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Expression and enzymatic activity of phenylalanine ammonia-lyase and p-coumarate 3-hydroxylase in mango (Mangifera indica 'Ataulfo') during ripening.

    PubMed

    Palafox-Carlos, H; Contreras-Vergara, C A; Muhlia-Almazán, A; Islas-Osuna, M A; González-Aguilar, G A

    2014-05-16

    Phenylalanine ammonia lyase (PAL) and p-coumarate 3-hydroxylase (C3H) are key enzymes in the phenylpropanoid pathway. The relative expression of PAL and C3H was evaluated in mango fruit cultivar 'Ataulfo' in four ripening stages (RS1, RS2, RS3, and RS4) by quantitative polymerase chain reaction. In addition, enzyme activity of PAL and C3H was determined in mango fruits during ripening. The PAL levels were downregulated at the RS2 and RS3 stages, while C3H levels were upregulated in fruits only at RS3. The enzyme activity of PAL followed a pattern that was different from that of the PAL expression, thus suggesting regulation at several levels. For C3H, a regulation at the transcriptional level is suggested because a similar pattern was revealed by its activity and transcript level. In this study, the complexity of secondary metabolite biosynthesis regulation is emphasized because PAL and C3H enzymes are involved in the biosynthesis of several secondary metabolites that are active during all mango ripening stages.

  19. Non-covalent Small-Molecule Kelch-like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 (Keap1-Nrf2) Inhibitors and Their Potential for Targeting Central Nervous System Diseases.

    PubMed

    Pallesen, Jakob S; Tran, Kim T; Bach, Anders

    2018-05-29

    The transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) has a protective effect against oxidative stress and plays a major role in inflammation and central nervous system (CNS) diseases. Inhibition of the protein-protein interaction (PPI) between Nrf2 and its repressor protein, Kelch-like ECH-associated protein 1 (Keap1), leads to translocation of Nrf2 from the cytosol to the nucleus and expression of detoxifying antioxidant enzymes. To date, several non-covalent small-molecule Keap1-Nrf2 inhibitors have been identified; however, many of them contain carboxylic acids and are rather large in size, which likely prevents or decreases CNS permeability. This Perspective describes current small-molecule Keap1-Nrf2 inhibitors with experimental evidence for the ability to inhibit the Keap1-Nrf2 interaction by binding to Keap1 in a non-covalent manner. Binding data, biostructural studies, and biological activity are summarized for the inhibitors, and their potential as CNS tool compounds is discussed by analyzing physicochemical properties, including CNS multiparameter optimization (MPO) scoring algorithms. Finally, several strategies for identifying CNS-targeting Keap1 inhibitors are described.

  20. Germination parameterization and development of an after-ripening thermal-time model for primary dormancy release of Lithospermum arvense seeds

    PubMed Central

    Chantre, Guillermo R.; Batlla, Diego; Sabbatini, Mario R.; Orioli, Gustavo

    2009-01-01

    Background and Aims Models based on thermal-time approaches have been a useful tool for characterizing and predicting seed germination and dormancy release in relation to time and temperature. The aims of the present work were to evaluate the relative accuracy of different thermal-time approaches for the description of germination in Lithospermum arvense and to develop an after-ripening thermal-time model for predicting seed dormancy release. Methods Seeds were dry-stored at constant temperatures of 5, 15 or 24 °C for up to 210 d. After different storage periods, batches of 50 seeds were incubated at eight constant temperature regimes of 5, 8, 10, 13, 15, 17, 20 or 25 °C. Experimentally obtained cumulative-germination curves were analysed using a non-linear regression procedure to obtain optimal population thermal parameters for L. arvense. Changes in these parameters were described as a function of after-ripening thermal-time and storage temperature. Key Results The most accurate approach for simulating the thermal-germination response of L. arvense was achieved by assuming a normal distribution of both base and maximum germination temperatures. The results contradict the widely accepted assumption of a single Tb value for the entire seed population. The after-ripening process was characterized by a progressive increase in the mean maximum germination temperature and a reduction in the thermal-time requirements for germination at sub-optimal temperatures. Conclusions The after-ripening thermal-time model developed here gave an acceptable description of the observed field emergence patterns, thus indicating its usefulness as a predictive tool to enhance weed management tactics. PMID:19332426