Effect of carrot puree edible films on quality preservation of fresh-cut carrots

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Wang X.

2015-06-01

Full Text Available The effect of edible films based on carrot puree, chitosan, corn starch, gelatin, glycerol and cinnamaldehyde on fresh-cut carrots was studied during storage. Several parameters, such as firmness, colour, weight loss, total carotenoids, total phenols, polyphenol oxidase (PPO activity and peroxidase (POD activity in coated carrots were determined at regular intervals and then compared with the uncoated carrots throughout the storage period. Significant and expected changes were observed in all carrot samples that were compared. The coating treatment significantly (P < 0.05 delayed the senescence, reduced the deterioration of exterior quality and retained total carotenoids well compared with control (P < 0.05. In addition, significant inhibition of PPO activity (P < 0.05 and POD activity (P < 0.05 as well as reduced accumulation of polyphenols (P < 0.05 were observed for all coated samples. All of these favourable responses induced by coating treatment on minimally processed fresh-cut carrots showed beneficial physiological effects, which would give some useful references to the fresh-cut fruit and vegetable processing industry and satisfy people’s requirements allowing for extending product shelf life without negatively affecting the sensory quality or acceptability.

Improved mechanical properties of retorted carrots by ultrasonic pre-treatments.

Science.gov (United States)

Day, Li; Xu, Mi; Øiseth, Sofia K; Mawson, Raymond

2012-05-01

The use of ultrasound pre-processing treatment, compared to blanching, to enhance mechanical properties of non-starchy cell wall materials was investigated using carrot as an example. The mechanical properties of carrot tissues were measured by compression and tensile testing after the pre-processing treatment prior to and after retorting. Carrot samples ultrasound treated for 10 min at 60 °C provided a higher mechanical strength (P<0.05) to the cell wall structure than blanching for the same time period. With the addition of 0.5% CaCl(2) in the pre-treatment solution, both blanching and ultrasound treatment showed synergistic effect on enhancing the mechanical properties of retorted carrot pieces. At a relatively short treatment time (10 min at 60 °C) with the use of 0.5% CaCl(2), ultrasound treatment achieved similar enhancement to the mechanical strength of retorted carrots to blanching for a much longer time period (i.e. 40 min). The mechanism involved appears to be related to the stress responses present in all living plant matter. However, there is a need to clarify the relative importance of the potential stress mechanisms in order to get a better understanding of the processing conditions likely to be most effective. The amount of ultrasound treatment required is likely to involve low treatment intensities and there are indications from the structural characterisation and mechanical property analyses that the plant cell wall tissues were more elastic than that accomplished using low temperature long time blanching. Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.

Plasmid-dependent attachment of Agrobacterium tumefaciens to plant tissue culture cells.

Science.gov (United States)

Matthysse, A G; Wyman, P M; Holmes, K V

1978-11-01

Kinetic, microscopic, and biochemical studies show that virulent Ti (tumor inducing)-plasmid-containing strains of Agrobacterium attach to normal tobacco and carrot tissue culture cells. Kinetic studies showed that virulent strains of A. tumefaciens attach to the plant tissue culture cells in increasing numbers during the first 1 to 2 h of incubation of the bacteria with the plant cells. Five Ti-plasmid-containing virulent Agrobacterium strains showed greater attachment to tobacco cells than did five avirulent strains. Light and scanning electron microscopic observations confirmed that virulent strains showed little attachment. Bacterial attachment was blocked by prior incubation of the plant cells with lipopolysaccharide extracted from A. tumefaciens, but not from A. radiobacter, suggesting that bacterial lipopolysaccharide is one of the components involved in the attachment process. At least one other bacterial product may be required for attachment in tissue culture because the virulent A. tumefaciens NT1, which lacks the Ti plasmid, does not itself attach to tobacco cells, but its lipopolysaccharide does inhibit the attachment of virulent strains.

The impact of Cu treatment on phenolic and polyamine levels in plant material regenerated from embryos obtained in anther culture of carrot

Czech Academy of Sciences Publication Activity Database

Górecka, K.; Cvikrová, Milena; Kowalska, U.; Eder, Josef; Szafrańska, K.; Górecki, R.; Janas, K. M.

2007-01-01

Roč. 45, č. 1 (2007), s. 54-61 ISSN 0981-9428 R&D Projects: GA MŠk 1P05OC052 Institutional research plan: CEZ:AV0Z50380511 Source of funding: V - iné verejné zdroje Keywords : carrot culture * copper ions * embryo regeneration Subject RIV: GE - Plant Breeding Impact factor: 1.669, year: 2007

Carrot cells: a pioneering platform for biopharmaceuticals production.

Science.gov (United States)

Rosales-Mendoza, Sergio; Tello-Olea, Marlene Anahí

2015-03-01

Carrot (Daucus carota L.) is of importance in the molecular farming field as it constitutes the first plant species approved to produce biopharmaceuticals for human use. In this review, features that make carrot an advantageous species in the molecular farming field are analyzed and a description of the developments achieved with this crop thus far is presented. A guide for genetic transformation procedures is also included. The state of the art comprises ten vaccine prototypes against Measles virus, Hepatitis B virus, Human immunodeficiency virus, Yersinia pestis, Chlamydia trachomatis, Mycobacterium tuberculosis, enterotoxigenic Escherichia coli, Corynebacterium diphtheria/Clostridium tetani/Bordetella pertussis, and Helicobacter pylori; as well as the case of the glucocerebrosidase, an enzyme used for replacement therapy, and other therapeutics. Perspectives for these developments are envisioned and innovations are proposed such as the use of transplastomic technologies-, hairy roots-, and viral expression-based systems to improve yields and develop new products derived from this advantageous plant species.

Process quality planning of quality function deployment for carrot syrup

Science.gov (United States)

Ekawati, Yurida; Noya, Sunday; Widjaja, Filemon

2017-06-01

Carrot products are rarely available in the market. Based on previous research that had been done using QFD to generate product design of carrots products, the research to produce the process quality planning had been carried out. The carrot product studied was carrot syrup. The research resulted in a process planning matrix for carrot syrup. The matrix gives information about critical process plan and the priority of the critical process plan. The critical process plan on the production process of carrot syrup consists of carrots sorting, carrots peeling, carrots washing, blanching process, carrots cutting, the making of pureed carrots, filtering carrot juice, the addition of sugar in carrot juice, the addition of food additives in carrot juice, syrup boiling, syrup filtering, syrup filling into the bottle, the bottle closure and cooling. The information will help the design of the production process of carrot syrup.

in Artificially Polluted Soil—Carrots System

Directory of Open Access Journals (Sweden)

Caroline Sablayrolles

2009-01-01

Full Text Available Surfactants are widely used in household and industrial products. The risk of incorporation of linear alkylbenzene sulfonates (LAS from biosolids, wastewater, and fertilizers land application to the food chain is being assessed at present by the European Union. In the present work, a complete analytical method for LAS trace determination has been developed and successfully applied to LAS (C10–C13 uptake in carrot plants used as model. These carrots were grown in soil with the trace organics compounds added directly into the plant containers in pure substances form. LAS trace determination (μg kg-1 dry matter in carrots samples was achieved by Soxtec apparatus and high-performance liquid chromatography-fluorescence detection. The methodology developed provides LAS determination at low detection limits (5 μg kg-1 dry matter for carrot sample (2 g dry matter with good recoveries rate (>90%. Transfer of LAS has been followed into the various parts of the carrot plant. LAS are generally found in the carrot leaves and percentage transfer remains very low (0.02%.

21 CFR 73.300 - Carrot oil.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Carrot oil. 73.300 Section 73.300 Food and Drugs... ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.300 Carrot oil. (a) Identity. (1) The color additive carrot oil is the liquid or the solid portion of the mixture or the mixture itself obtained by the hexane...

The loss of plasma membrane lysopip and an increase of PIP2 result from treatment of carrot cells with fungal enzymes

International Nuclear Information System (INIS)

Chen, Q.; Boss, W.F.

1989-01-01

The plasma membranes of carrot cells grown in suspension culture are enriched with PIP, lysoPIP, and PIP 2 . To determine whether or not these lipids are involved in signal transduction, we have challenged the cells with a mixture of fungal cellulases, Driselase, and monitored the changes in the phosphoinositides and in the phosphoinositide kinase activity. With cell prelabeled with [ 3 H]inositol, two major changes are observed: (1) lysoPIP decreases 30% compared to the sorbitol control and (2) PIP 2 doubles. There is no increase in IP, IP 2 , or IP 3 . In vitro phosphorylation studies using [γ- 32 P]ATP indicate that the increase in PIP 2 is due in part to activation of the PIP kinase. These data suggest that the role of the polyphosphoinositides in signal transduction in plants may involve activation of the PIP kinase and/or activation of A type phospholipases rather than C type phospholipases

Isolation of carrot plant lines with altered carotene contents from gamma irradiated explants

International Nuclear Information System (INIS)

Pawlicki, N.; Sangwan, R.S.; Sangwan-Norreel, B.S.

2001-01-01

Dietary vitamin A is mainly obtained from carotenes of vegetables and fruits. Carrot (Daucus carota L.) is one of the major sources of carotene. Carrot cultivars have been obtained mainly through classical breeding, and genetic selection has permitted the creation of new varieties with high carotene contents. The fact that in several crops agronomically important mutants/variants have been generated by in vitro culture techniques prompted us to combine gamma irradiation and in vitro somatic embryogenesis to obtain regenerants with variations in carotene content in carrot. To test the effect of gamma rays on somatic embryogenesis and on the carotene level, aseptically germinated seedlings of 8 carrot varieties were exposed to 5; 10 and 500 Gy before culturing petiole segments on LN1 medium. Non-irradiated petioles produced calli with somatic embryos, while irradiated explants reacted differently according to radiation dose. After 4 weeks of culture on LN1 medium, petiole segments of different varieties irradiated with 5 and 10 Gy gave more callus with embryos than those with non-irradiated segments. However, after the subculture on LN medium, the development of embryos into plantlets was rare. It was also noted that after irradiation with 5 Gy, the petiole segments gave voluminous calli. Further, in variety 'Chantenay', the irradiated calli were deep orange while non-irradiated calli were green. However, embryo formation was not observed in these calli. This orange coloration suggests an appreciable synthesis of carotene in the calli. Gamma rays, probably produced cell lines with different colors and carotene content. Of the 8 cultivars tested, normal plantlets of 3 varieties were regenerated from somatic embryos irradiated with 10 Gy, and were transferred to greenhouse to develop roots. For each assay, the carotene analysis was carried out on 2 roots, and compared with plants produced from non-irradiated somatic embryos. Carotene level in the plants, derived from

Pulsed electric field improves the bioprotective capacity of purées for different coloured carrot cultivars against H2O2-induced oxidative damage.

Science.gov (United States)

Leong, Sze Ying; Oey, Indrawati; Burritt, David John

2016-04-01

This research aimed to study the effect of pulsed electric field (PEF) processing on the bioprotective capacity of carrot purée for White Belgian, Yellow Solar, Nantes, Nutri Red and Purple Haze cultivars against H2O2-induced oxidative damage. The bioprotective capacity was determined using cell viability, membrane integrity and nitric oxide (NO) production in a human Caco-2 cell culture assay. Total carotenoids, total anthocyanins, total vitamin C and total phenolics were also evaluated. Compared to the untreated purée, Purple Haze and Nutri Red processed at 303 kJ/kg completely increased Caco-2 cells resistance towards oxidative damage by recovering the cell viability and inhibiting NO production. For cultivar with low carotenoid levels, i.e. Yellow Solar, the application of 0.8 kV/cm resulted in a higher total carotenoid content in the purée than its untreated counterpart, leading to an improved bioprotective effect. This study clearly shows that PEF could add value to carrots by maximising bioprotective effects. Copyright © 2015 Elsevier Ltd. All rights reserved.

Identification and characterization of bZIP-type transcription factors involved in carrot (Daucus carota L.) somatic embryogenesis.

Science.gov (United States)

Guan, Yucheng; Ren, Haibo; Xie, He; Ma, Zeyang; Chen, Fan

2009-10-01

Seed dormancy is an important adaptive trait that enables seeds of many species to remain quiescent until conditions become favorable for germination. Abscisic acid (ABA) plays an important role in these developmental processes. Like dormancy and germination, the elongation of carrot somatic embryo radicles is retarded by sucrose concentrations at or above 6%, and normal growth resumes at sucrose concentrations below 3%. Using a yeast one-hybrid screening system, we isolated two bZIP-type transcription factors, CAREB1 and CAREB2, from a cDNA library prepared from carrot somatic embryos cultured in a high-sucrose medium. Both CAREB1 and CAREB2 were localized to the nucleus, and specifically bound to the ABA response element (ABRE) in the Dc3 promoter. Expression of CAREB2 was induced in seedlings by drought and exogenous ABA application; whereas expression of CAREB1 increased during late embryogenesis, and reduced dramatically when somatic embryos were treated with fluridone, an inhibitor of ABA synthesis. Overexpression of CAREB1 caused somatic embryos to develop slowly when cultured in low-sucrose medium, and retarded the elongation of the radicles. These results indicate that CAREB1 and CAREB2 have similar DNA-binding activities, but play different roles during carrot development. Our results indicate that CAREB1 functions as an important trans-acting factor in the ABA signal transduction pathway during carrot somatic embryogenesis.

Advances in transforming kudzu (Pueraria phaseoloides and carrot (Daucus carota var. Danvers 126 roots with different Agrobacterium rhizogenes strains for increasing MA fungi growth

Directory of Open Access Journals (Sweden)

Marisol Medina Sierra

2002-07-01

Full Text Available Kudzú (P. phaseoloides and carrot (D. carota roots were transformed in this survey into different kinds of culture medium by using five different A. rhizogenes strains. These presented different behaviour both in carrot transformation by A. rhizogenes 15834, A.r.8196 and A.r.2659 strains as well as kudzu transformation by A.r.15834 and A.r.1724 strains. Transformed carrot root growth was increased in WM culture medium, whilst transformed kudzu root growth did not increase in either the same medium or in modified MS medium. Transformed carrot roots were used for G. intrarradices increase and sporulation; however, wild AMF strains, isolated from a mining area (the lower Cauca area of Antioquia, did not grow either in roots from this specie or those from kudzu, in spite of this plant having great affinity for wild AMF strains. The results represent an advance in the procedure for DNA isolation and keeping AMF collections, required for other research.

Solid Loss of Carrots During Simulated Gastric Digestion.

Science.gov (United States)

Kong, Fanbin; Singh, R Paul

2011-03-01

The knowledge of solid loss kinetics of foods during digestion is crucial for understanding the factors that constrain the release of nutrients from the food matrix and their fate of digestion. The objective of this study was to investigate the solid loss of carrots during simulated gastric digestion as affected by pH, temperature, viscosity of gastric fluids, mechanical force present in stomach, and cooking. Cylindrical carrot samples were tested by static soaking method and using a model stomach system. The weight retention, moisture, and loss of dry mass were determined. The results indicated that acid hydrolysis is critical for an efficient mass transfer and carrot digestion. Internal resistance rather than external resistance is dominant in the transfer of soluble solids from carrot to gastric fluid. Increase in viscosity of gastric fluid by adding 0.5% gum (w/w) significantly increased the external resistance and decreased mass transfer rate of carrots in static soaking. When mechanical force was not present, 61% of the solids in the raw carrot samples were released into gastric fluid after 4 h of static soaking in simulated gastric juice. Mechanical force significantly increased solid loss by causing surface erosion. Boiling increased the disintegration of carrot during digestion that may favor the loss of solids meanwhile reducing the amount of solids available for loss in gastric juice. Weibull function was successfully used to describe the solid loss of carrot during simulated digestion. The effective diffusion coefficients of solids were calculated using the Fick's second law of diffusion for an infinite cylinder, which are between 0.75 × 10(-11) and 8.72 × 10(-11) m(2)/s, depending on the pH of the gastric fluid.

Pathogenic mycoflora on carrot seeds

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Bogdan Nowicki

2013-12-01

Full Text Available Altogether 300 seed samples were collected during 9 years in 8 regions of Poland and the fungi Were isolated and their pathogenicity to carrot seedlings was examined. Alternaria rudicina provcd to be the most important pathogen although. A. alternata was more common. The other important pathogens were Fusarium spp., Phoma spp. and Botrytis cinerea. The infection of carrot seeds by A. radicina should be used as an important criterium in seed quality evaluation.

INFLUENCE OF MEDIA GELLING AGENTS ON ROOT BIOMASS AND IN VITRO VA-MYCORRfflZAL SYMBIOSIS OF CARROT WITH GIGASPORA MARGARITA

Directory of Open Access Journals (Sweden)

ANJALI VERMA

1997-01-01

Full Text Available An in vitro study with Ri-TDNA transformed roots of carrot (Daucus carota was carried out to evaluate the role of macro-elements contributed as impurities in the gelling agent (phytagel over and above those present in the minimal (M medium. Production of root biomass was taken as a measure to quantify the influence of macro-elements added to the minimal medium. The levels of phosphorus when adjusted to 1.19 mg/1 and 1.09 mg/l, lead to dry root biomass production at par with the control. Attempts made to lower the amount of impurities in phytagel by de-ionization using different alkalies, proved NaOH to give the best results in terms of relatively high amount of root biomass. In an in vitro dual culture system with carrot as host and Gigaspora margarita as the vesicular-arbuscular mycorrhizal fungus, phytagel impurities helped to produce maximum number of infection units and auxiliary cells when phytagel was added to the minimal medium.

Calmodulin stimulation of calcium transport in carrot microsomal vesicles

International Nuclear Information System (INIS)

Pierce, W.S.; Sze, H.

1987-01-01

ATP-dependent 45 Ca 2+ uptake into microsomal vesicles isolated from cultured carrot cells (Daucus carota Danvers) was stimulated 2-3 fold by 5 ug/ml calmodulin (CaM). Microsomal vesicles separated with a linear sucrose gradient showed two peaks with CaM-stimulated Ca 2+ uptake activities. One peak (at 1.12 g/cc) comigrated with the activity of the antimycin A-insensitive NADH-dependent cytochrome c reductase. This transport activity was enhanced 10-20 fold by 10 mM oxalate and appeared to be associates with vesicles derived primarily from the ER. The other peak of CaM-stimulated Ca 2+ uptake (at 1.17 g/cc) was not affected by oxalate. These vesicles are probably derived from the plasma membrane. Preliminary experiments with the low-density vesicles (ER) vesicles, indicate that inositol-1,4,5-trisphosphate caused a transient reduction in intravesicular Ca 2+ . These results are consistent with the ER being an important site of intracellular Ca 2+ regulation

Soil solarization for weed control in carrot

Directory of Open Access Journals (Sweden)

MARENCO RICARDO ANTONIO

2000-01-01

Full Text Available Soil solarization is a technique used for weed and plant disease control in regions with high levels of solar radiation. The effect of solarization (0, 3, 6, and 9 weeks upon weed populations, carrot (Daucus carota L. cv. Brasília yield and nematode infestation in carrot roots was studied in São Luís (2º35' S; 44º10' W, MA, Brazil, using transparent polyethylene films (100 and 150 mm of thickness. The maximum temperature at 5 cm of depth was about 10ºC warmer in solarized soil than in control plots. In the study 20 weed types were recorded. Solarization reduced weed biomass and density in about 50% of weed species, including Cyperus spp., Chamaecrista nictans var. paraguariensis (Chod & Hassl. Irwin & Barneby, Marsypianthes chamaedrys (Vahl O. Kuntze, Mitracarpus sp., Mollugo verticillata L., Sebastiania corniculata M. Arg., and Spigelia anthelmia L. Approximately 40% of species in the weed flora were not affected by soil mulching. Furthermore, seed germination of Commelina benghalensis L. was increased by soil solarization. Marketable yield of carrots was greater in solarized soil than in the unsolarized one. It was concluded that solarization for nine weeks increases carrot yield and is effective for controlling more than half of the weed species recorded. Mulching was not effective for controlling root-knot nematodes in carrot.

Drying of carrots in slices with osmotic dehydration

African Journals Online (AJOL)

SAM

2014-07-23

Jul 23, 2014 ... extend the shelf-life by a few weeks, one year or more. The methods .... drated carrots, this work studied the drying of carrot with pre-osmotic ... e) Weight Loss - obtained directly using balance semi-analytical model BEL ...

Component-resolved in vitro diagnosis of carrot allergy in three different regions of Europe

DEFF Research Database (Denmark)

Ballmer-Weber, B K; Skamstrup Hansen, K; Sastre, J

2012-01-01

Carrot is a frequent cause of food allergy in Europe. The objective of this study was to evaluate a panel of carrot allergens for diagnosis of carrot allergy in Spain, Switzerland and Denmark.......Carrot is a frequent cause of food allergy in Europe. The objective of this study was to evaluate a panel of carrot allergens for diagnosis of carrot allergy in Spain, Switzerland and Denmark....

Keeping carrots quality during storage by irradiation

International Nuclear Information System (INIS)

El-Nashaby, F.M.; Hafez, S.A.

2000-01-01

Fresh carrots were gamma irradiated at 0, 0.1, 0.2, 0.5 and 1.0 KGy doses using cobalt-60 source. A portion of the irradiated carrots was stored at ambient temperature (20-25 degree C) and the rest was packed in polyethylene bags and stored in refrigerator temperature (5± 1 degree C). The results indicated that the sprouting of carrots were completely inhibited by radiation dose of 0.2 KGy either at ambient temperature or cold storage, while the non-irradiated samples started to sprout after 2 and 4 weeks of storage at ambient temperature and cold storage respectively. Furthermore, samples treated with gamma rays showed lower values of weight loss compared to the untreated ones during storage. Concerning storage temperature, it was found that the rate of weight loss during storage was higher at ambient temperature than in refrigerator. Micro-organisms were greatly affected by gamma rays, their counts were found to be decreased by increasing irradiation dose as compared with the control sample. Moreover, during storage period, refrigerated samples showed lower bacterial and molds counts than samples stored at ambient temperature. On the other hand, it was also observed that irradiated samples at the higher dose i.e. 1.0 KGy stored either at ambient temperature or in refrigerator showed higher bacterial and molds counts than unirradiated ones. The lowest dose (0.1 KGy) had no effect on carotene and sugars contents of carrots while increasing the dose above 0.2 KGy led to gradual decrease in carotene contents and gradual increase in total soluble sugars contents of the samples with increasing the irradiation dose compared with the nonirradiated ones. The results proved that gamma irradiation dose of 0.2 Gy was effective for sprout inhibition of carrots and increase their shelf-life to 10 weeks at ambient temperature without adverse effects on carotene and sugars contents of treated carrots

Carrot juice ingestion attenuates high fructose-induced circulatory pro-inflammatory mediators in weanling Wistar rats.

Science.gov (United States)

Mahesh, Malleswarapu; Bharathi, Munugala; Raja Gopal Reddy, Mooli; Pappu, Pranati; Putcha, Uday Kumar; Vajreswari, Ayyalasomayajula; Jeyakumar, Shanmugam M

2017-03-01

Adipose tissue, an endocrine organ, plays a vital role not only in energy homeostasis, but also in the development and/or progression of various metabolic diseases, such as insulin resistance, type 2 diabetes and non-alcoholic fatty liver disease (NAFLD), via several factors and mechanisms, including inflammation. This study tested, whether carrot juice administration affected the adipose tissue development and its inflammatory status in a high fructose diet-induced rat model. For this purpose, male weanling Wistar rats were divided into four groups and fed either control or high fructose diet of AIN-93G composition with or without carrot juice ingestion for an 8 week period. Administration of carrot juice did not affect the adiposity and cell size of visceral fat depot; retroperitoneal white adipose tissue (RPWAT), which was corroborated with unaltered expression of genes involved in adipogenic and lipogenic pathways. However, it significantly reduced the high fructose diet-induced elevation of plasma free fatty acid (FFA) (P ≤ 0.05), macrophage chemoattractant protein 1 (MCP1) (P ≤ 0.01) and high sensitive C-reactive protein (hsCRP) (P ≤ 0.05) levels. Carrot juice administration attenuated the high fructose diet-induced elevation of levels of circulatory FFA and pro-inflammatory mediators; MCP1 and hsCRP without affecting the adiposity and cell size of visceral fat depot; RPWAT. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

growth and yield response of carrot (daucus carota l.)

African Journals Online (AJOL)

User

Mampong Campus to investigate the growth and yield responses of carrot to different rates of soil amendments ... quires a deep and well-drained loamy soil with high amount of ... The factors considered in the ..... processing carrot root yields.

Uptake of 13C-glucose by cell suspensions of carrot (Daucus carota) measured by in vivo NMR: Cycling of triose, pentose- and hexose-phosphates

NARCIS (Netherlands)

Krook, J.; Vreugdenhil, D.; Dijkema, C.; Plas, van der L.H.W.

2000-01-01

After a lag phase of 2 days, batch-grown cells of carrot (Daucus carota L.) cv. Flakkese entered the exponential growth phase and started to accumulate sucrose and hexoses. Short-term feeding 13C-glucose in this period resulted in only minor labelling of sucrose or fructose. CO2 production from

Production of Bioethanol from Carrot Pomace Using the Thermotolerant Yeast Kluyveromyces marxianus

Energy Technology Data Exchange (ETDEWEB)

Chi-Yang Yu; Bo-Hong Jiang; Kow-Jen Duan [Tatung University, Tapei, Taiwan (China). Department of Bioengineering

2013-03-15

Carrot pomace, a major agricultural waste from the juice industry, was used as a feedstock for bioethanol production by fermentation with the thermotolerant yeast Kluyveromyces marxianus. Treatment of the carrot pomace with Accellerase(TM) 1000 and pectinase at 50 °C for 84 h, resulted in conversion of 42% of its mass to fermentable sugars, mainly glucose, fructose, and sucrose. Simultaneous saccharification and fermentation (SSF) at 42 °C was performed on 10% (w/v) carrot pomace; the concentration of ethanol reached 18 g/L and the yield of ethanol from carrot pomace was 0.18 g/g. The highest ethanol concentration of 37 g/L was observed with an additional charge of 10% supplemented to the original 10% of carrot pomace after 12 h; the corresponding yield was 0.185 g/g. Our results clearly demonstrated the potential of combining a SSF process with thermotolerant yeast for the production of bioethanol using carrot pomace as a feedstock.

Release of somatic embryogenic potential from excised zygotic embryos of carrot and maintenance of proembryonic cultures in hormone-free medium

Science.gov (United States)

Smith, D. L.; Krikorian, A. D.

1989-01-01

Excised zygotic embryos, mericarps ("seeds") and hypocotyls of seedlings of cultivated carrot Daucus carota cv. Scarlet Nantes were evaluated for their ability to generate somatic embryos on a semisolid hormone-free nutrient medium. Neither intact zygotic embryos nor hypocotyls ever produced somatic embryos. However, mericarps and broken zygotic embryos were excellent sources for somatic embryo production (response levels as high as 86%). Somatic embryo formation was highest from cotyledons, but was also observed on isolated hypocotyls and root tips of mature zygotic embryos. On media containing unreduced nitrogen, somatic embryo formation led to the generation of vigorous cultures comprised entirely of somatic embryos at various stages of development which in turn proliferated still other somatic embryos. However, a medium was devised which when 1-5 mM NH4+ was the sole nitrogen source, led only to a proliferation of globular proembryos. Sustained subculturing of these proembryos at 2-3 week intervals enabled establishment of highly uniform cultures in which no further development into more mature stages of embryonic development occurred. These have been maintained, without decline, as morphogenetically competent proembryonic globules for over ten months. A basal medium containing from 1-5 mM NH4+ as the sole nitrogen source appears not to be inductive to somatic proembryo formation. Instead, such a medium is best thought of as permissive to the expression of embryogenically determined cells within zygotic embryos. By excising and breaking or wounding zygotic embryos, constituent cells are probably released from positional or chemical restraints and thus are able to express their innate embryogenic potential. Once a proembryonic culture is established, this medium containing 1-5 mM NH4+ as the sole nitrogen source provides a nonpermissive environment to the development and growth of later embryonic stages, but it does allow the continued formation and

Some microbiological, histopathological and biochemical changes in the broiler chicken fed carrot leaves

International Nuclear Information System (INIS)

El-Faramawy, A.A.; Soliman, S.M.; Fahmy, M.O.

2005-01-01

Ninety 21 day old Arbor Acres chicks arranged into 6 equal groups were used in the present study to evaluate the effect of substitution of 10% yellow corn (YC) with berseem, green carrot leaves without and with digestive enzymatic mixture, untreated dry carrot leaves and treated ones with Aspergillus niger on the intestinal microflora, histopathology of liver and intestine in addition to some biochemical parameters. Replacement of (YC) with green carrot leaves, untreated and treated carrot leaves caused significant (P<0.05) increase in total bacterial count and significant (P<0.05) decrease in Lactobacilli, while the addition of enzyme to the green carrot leaves led to decrease in total bacterial count and increase in Lactobacillus count. The histopathological changes of both intestine and liver were restricted only to both groups of dried carrot leaves. Similarly untreated and treated dry carrot leaves groups were also associated with significant increase in triglyceride, ALT and significant (P<0.05) decrease in cholesterol in serum. From the above mentioned results, it could be concluded that dried carrot leaves, untreated or treated with Aspergillus niger, were unsuitable for broiler chicken feeding since they directly affected bird's gut health causing change in nutrient absorption and considerable decrease in the beneficial bacterial count

Postharvest quality of carrot cultivars, packaged and in bulk

African Journals Online (AJOL)

Jane

2011-08-15

Aug 15, 2011 ... presented the highest number of sprouts when stored in bulk (48.5%) and the largest number of carrots with radicels (54.7%). ... The carrot (Daucus carota L.), a plant of the family. Apiaceae ... widespread global consumption and large planted area. .... increased temperature and CO2 concentration and the.

The effects of ridging, row-spacing and seeding rate on carrot yield

Directory of Open Access Journals (Sweden)

S. TAIVALMAA

2008-12-01

Full Text Available Cool, wet spring weather often delays the early growth of carrots (Daucus carota L. in northern Europe. This effect may be partly obviated by sowing in ridges. Many types of ridges are used, but the most suitable for carrot cultivation under the conditions prevailing in northern Europe has yet to be determined. The effects of ridging, seeding rate and sowing system on the yield and visible quality of carrots were therefore studied in the field during three years. The highest yields were recorded for carrots sown in double rows on a narrow ridge. The effect of sowing system on mean root weight differed depending on the ridging regime. The mean weight of roots was higher for carrots cultivated on broad ridges than in other systems. Seeding rate had the most significant effect on mean root weight. For industrial purposes it is recommended that carrots be cultivated on broad ridges in double rows at low seeding rates with irrigation. The optimal cultivation technique for carrots destined for the fresh vegetable market would be narrow ridges sown in double rows at high seeding rates. The ridging system, seeding rate and row spacing did not appear to affect the external quality of roots. More detailed studies should be carried out to establish the effects of abiotic growth factors under different ridging regimes.;

The effects of ridging, row-spacing and seeding rate on carrot yield

Directory of Open Access Journals (Sweden)

Sanna-Liisa Taivalmaa

1997-12-01

Full Text Available Cool, wet spring weather often delays the early growth of carrots (Daucus carota L. in northern Europe. This effect may be partly obviated by sowing in ridges. Many types of ridges are used, but the most suitable for carrot cultivation under the conditions prevailing in northern Europe has yet to be determined. The effects of ridging, seeding rate and sowing system on the yield and visible quality of carrots were therefore studied in the field during three years. The highest yields were recorded for carrots sown in double rows on a narrow ridge. The effect of sowing system on mean root weight differed depending on the ridging regime. The mean weight of roots was higher for carrots cultivated on broad ridges than in other systems. Seeding rate had the most significant effect on mean root weight. For industrial purposes it is recommended that carrots be cultivated on broad ridges in double rows at low seeding rates with irrigation. The optimal cultivation technique for carrots destined for the fresh vegetable market would be narrow ridges sown in double rows at high seeding rates. The ridging system, seeding rate and row spacing did not appear to affect the external quality of roots. More detailed studies should be carried out to establish the effects of abiotic growth factors under different ridging regimes.

Transplacental inhibitory effect of carrot juice on the clastogenicity of cyclophosphamide in mice

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Gimmler-Luz Maria Clara

1999-01-01

Full Text Available Genetic damage during the prenatal period can provoke important neoplastic alterations and other diseases in postnatal life. Beta-carotene (ßC is considered to be one of the most important anticarcinogens in the diet and can protect mammalian cells against genotoxic events. As carrots are important dietary source of ßC, we decided to test the effect of fresh carrot juice (CaJ on cyclophosphamide (CP-induced genotoxicity in maternal and fetal erythropoietic tissues. The treatment with CaJ started on the 7th day of the pregnancy of BALB/c female mice. We observed, on the 16th gestational day, that this treatment did not modify the spontaneous frequency of micronucleated polychromatic erythrocytes (mPCE in the bone marrow of the females nor in the livers of their fetuses. The mPCE frequency observed 24 h after an intraperitoneal injection of CP (40 mg/kg on the 15th day was significantly lower in CaJ-pretreated pregnant female bone marrow and in the liver of their fetuses than those observed in the group treated with CP only. These results demonstrate the presence of natural anticlastogens in carrots.

SE-ENRICHMENT OF CARROT AND ONION VIA FOLIAR APPLICATION

OpenAIRE

Kapolna, Emese; Laursen, Kristian H.; Hillestrøm, Peter; Husted, Søren; Larsen, Erik H.

2008-01-01

The aim of this work was to study the selenium accumulation in carrot and onion plants using foliar application by sodium selenite and sodium selenate. Furthermore, we aimed at identifying the Se species biosynthesised by onion and carrot plants. The results were used to prepare for production of 77Se enriched plants for an ongoing human absorption study.

Hydrogen production from carrot pulp by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

NARCIS (Netherlands)

Vrije, de G.J.; Budde, M.A.W.; Lips, S.J.J.; Bakker, R.R.; Mars, A.E.; Claassen, P.A.M.

2010-01-01

Hydrogen was produced from carrot pulp hydrolysate, untreated carrot pulp and (mixtures of) glucose and fructose by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana in pH-controlled bioreactors. Carrot pulp hydrolysate was obtained after enzymatic hydrolysis

Characterization of centromeric histone H3 (CENH3 variants in cultivated and wild carrots (Daucus sp..

Directory of Open Access Journals (Sweden)

Frank Dunemann

Full Text Available In eukaryotes, centromeres are the assembly sites for the kinetochore, a multi-protein complex to which spindle microtubules are attached at mitosis and meiosis, thereby ensuring segregation of chromosomes during cell division. They are specified by incorporation of CENH3, a centromere specific histone H3 variant which replaces canonical histone H3 in the nucleosomes of functional centromeres. To lay a first foundation of a putative alternative haploidization strategy based on centromere-mediated genome elimination in cultivated carrots, in the presented research we aimed at the identification and cloning of functional CENH3 genes in Daucus carota and three distantly related wild species of genus Daucus varying in basic chromosome numbers. Based on mining the carrot transcriptome followed by a subsequent PCR-based cloning, homologous coding sequences for CENH3s of the four Daucus species were identified. The ORFs of the CENH3 variants were very similar, and an amino acid sequence length of 146 aa was found in three out of the four species. Comparison of Daucus CENH3 amino acid sequences with those of other plant CENH3s as well as their phylogenetic arrangement among other dicot CENH3s suggest that the identified genes are authentic CENH3 homologs. To verify the location of the CENH3 protein in the kinetochore regions of the Daucus chromosomes, a polyclonal antibody based on a peptide corresponding to the N-terminus of DcCENH3 was developed and used for anti-CENH3 immunostaining of mitotic root cells. The chromosomal location of CENH3 proteins in the centromere regions of the chromosomes could be confirmed. For genetic localization of the CENH3 gene in the carrot genome, a previously constructed linkage map for carrot was used for mapping a CENH3-specific simple sequence repeat (SSR marker, and the CENH3 locus was mapped on the carrot chromosome 9.

Identification of measures for prevention of black spots in organically produced stored carrots

NARCIS (Netherlands)

Köhl, J.; Kastelein, P.; Elderson, J.; Blok, W.J.

2005-01-01

In the Netherlands winter carrot for the fresh market is a cash crop for many organic growers of field vegetables. In the last few years blackish spots have been observed during storage of carrot roots. An inventory was carried out in 2001/2002 and 2002/2003 including in total 42 carrot lots from 22

Solid Loss of Carrots During Simulated Gastric Digestion

OpenAIRE

Kong, Fanbin; Singh, R. Paul

2010-01-01

The knowledge of solid loss kinetics of foods during digestion is crucial for understanding the factors that constrain the release of nutrients from the food matrix and their fate of digestion. The objective of this study was to investigate the solid loss of carrots during simulated gastric digestion as affected by pH, temperature, viscosity of gastric fluids, mechanical force present in stomach, and cooking. Cylindrical carrot samples were tested by static soaking method and using a model st...

Evaluation the Sensory and Probiotics Properties of the Yogurt Supplemented with Carrot Juice

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Carmen R. Pop

2015-11-01

Full Text Available This study aimed to develop a new type of yogurt through addition of carrot juice. Yogurt is a fermented milk product obtained from fermentation of Lactobacillus bulgaricus and Streptococcus thermophilus strains. The effect of carrot juice at different levels (0%, 8%, 16%, 24% on sensory properties and the viability of probiotic bacteria in yogurts during storage (21 days at refrigerated temperature (4°C was evaluated. The yogurt supplementation with 24 % carrot juice significantly improves the stability of the lactic acid bacteria, that contained the recommended levels of 107 cfu/g probiotic bacteria at the end of 21-days shelf life. The yogurt with 24% carrot juice was the most appreciated (7.07 points, followed by the classic yogurt (6 points, yogurt with 8% (5.28 points and yogurt with16% carrot juice (5.5 points.

NM-Scale Anatomy of an Entire Stardust Carrot Track

Science.gov (United States)

Nakamura-Messenger, K.; Keller, L. P.; Clemett, S. J.; Messenger, S.

2009-01-01

Comet Wild-2 samples collected by NASA s Stardust mission are extremely complex, heterogeneous, and have experienced wide ranges of alteration during the capture process. There are two major types of track morphologies: "carrot" and "bulbous," that reflect different structural/compositional properties of the impactors. Carrot type tracks are typically produced by compact or single mineral grains which survive essentially intact as a single large terminal particle. Bulbous tracks are likely produced by fine-grained or organic-rich impactors [1]. Owing to their challenging nature and especially high value of Stardust samples, we have invested considerable effort in developing both sample preparation and analytical techniques tailored for Stardust sample analyses. Our report focuses on our systematic disassembly and coordinated analysis of Stardust carrot track #112 from the mm to nm-scale.

The effect of slicing type on drying kinetics and quality of dried carrot

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M Naghipour zadeh mahani

2016-04-01

, wavy blade and Ridged blade and the cutting direction at 3 levels (linear, lateral and diagonal were evaluated on drying kinetics, drying rate, shrinkage and rehydration. Statistic analysis done by SPSS software. Results and Discussion: The results of analysis of variance showed that the effects of cutting parameters were significant on studied parameters (p<0.01 (Table 1. Thin layers dried faster than thick layers because of firmness of surface which it causes slow moisture transfer. The least drying time was 200 minutes at the samples that cut by a wavy blade at the lateral direction with a significant difference (p<0.05 given Fig.3. In these samples surface evaporation is more, because of more surface. The compare means showed drying rate at thick layer is fewer because of the longer distance moisture removal (Fig.6. Also the most drying rate was 0.74 gmin-1 at cutting by flat blade on linear direction with a significant difference (p<0.05.The least shrinkage was obtained on this treatment was 36.7% given Fig.8. The most of tissue of linear slices is woody part that is dense compare with other parts therefore shrinkage decrease at during drying. The most rehydration was 3.96 and 3.88 for cutting by flat blade in diagonal and linear direction with significant difference to other treatments. Rehydration depends on cell damage greatly. Since the slices of carrot that cut by flat blade were damaged fewer than other treatments therefore rehydration was more. Conclusions: The drying behavior of carrot slices was studied at different methods in slicing carrot. The results showed a significant effect of the cutting variables on drying kinetics, drying rate, shrinkage and rehydration. The carrot moisture content decreases continuously over the drying and the fastest drying occurred at thin layers sliced by wavy blade. The slices that were cut by flat blade at linear direction caused the best quality. The results show cutting parameters are significant effect on quality of

The Role of Pectin in Pb Binding by Carrot Peel Biosorbents: Isoterm Adsorption Study

Science.gov (United States)

Hastuti, B.; Totiana, F.; Winiasih, R.

2018-04-01

Cheaply and abundantly biosorption available materials such as carrot peels can be a cost-efficient method for removing heavy metals from wastewater. To investigate the role pectin plays in metal binding by carrot peels, commerce pectin was compared. FTIR spectra confirmed the presence of carboxyl and hydroxyl groups in commerce pectin and carrot pectin. Isoterm experiments showed that all materials could remove Pb (II) ion. All of materials binding Pb (II) follow Freundlich models adsorption. The commerce pectin bindsPb (II) by involving energy 16.6 KJ/mole whereas pectin from carrot peel involves energy 21.09 KJ/mole. It indicates that commerce pectin binds the Pb (II) by physics adsorption whereas pectin from carrot peel by physics and chemical adsorption.

Hydrogen production from carrot pulp by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

Energy Technology Data Exchange (ETDEWEB)

Vrije, Truus de; Budde, Miriam A.W.; Lips, Steef J.; Bakker, Robert R.; Mars, Astrid E.; Claassen, Pieternel A.M. [Wageningen UR, Food and Biobased Research, P.O. Box 17, 6700 AA Wageningen (Netherlands)

2010-12-15

Hydrogen was produced from carrot pulp hydrolysate, untreated carrot pulp and (mixtures of) glucose and fructose by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana in pH-controlled bioreactors. Carrot pulp hydrolysate was obtained after enzymatic hydrolysis of the polysaccharide fraction in carrot pulp. The main sugars in the hydrolysate were glucose, fructose, and sucrose. In fermentations with glucose hydrogen yields and productivities were similar for both strains. With fructose the hydrogen yield of C. saccharolyticus was reduced which might be related to uptake of glucose and fructose by different types of transport systems. With T. neapolitana the fructose consumption rate and consequently the hydrogen productivity were low. The hydrogen yields of both thermophiles were 2.7-2.8 mol H{sub 2}/mol hexose with 10 g/L sugars from carrot pulp hydrolysate. With 20 g/L sugars the yield of T. neapolitana was 2.4 mol H{sub 2}/mol hexose while the yield of C. saccharolyticus was reduced to 1.3 mol H{sub 2}/mol hexose due to high lactate production in the stationary growth phase. C. saccharolyticus was able to grow on carrot pulp and utilized soluble sugars and, after adaptation, pectin and some (hemi)cellulose. No growth was observed with T. neapolitana when using carrot pulp in agitated fermentations. Enzymatic hydrolysis of the polysaccharide fraction prior to fermentation increased the hydrogen yield with almost 10% to 2.3 g/kg of hydrolyzed carrot pulp. (author)

Improving microbiological safety and maintaining sensory and nutritional quality of pre-cut tomato and carrot by gamma irradiation

International Nuclear Information System (INIS)

Mohácsi-Farkas, Cs.; Nyirő-Fekete, B.; Daood, H.; Dalmadi, I.; Kiskó, G.

2014-01-01

Pre-cut tomato and carrot were irradiated with doses of 1.0, 1.5 and 2 kGy. Unirradiated control and irradiated samples were compared organoleptically by a sensory panel. Microbiological analyses were performed directly after irradiation and during post-irradiation storage for 8 days at 5 °C. Ascorbic acid contents, composition of carotenoids and tocopherols were determined. Statistically significant differences of sensory scores between unirradiated and irradiated samples were observed only in the texture of sliced carrots. Total aerobic viable cell counts have been reduced by about two log cycles with 1.5 kGy dose. Total coliforms and moulds were below the detection limit of 15 CFU/g in the irradiated samples during the refrigerated storage. Yeasts were relatively resistant part of the microbiota of pre-cut tomatoes, but 2 kGy dose reduced them below the detection limit. In pre-cut tomatoes, alpha-tocopherol and some carotenoids seemed to be the most radio-sensitive losing approximately one-third of their original concentrations at the dose of 2 kGy. At this dose tocopherols and the level of ascorbic acid decreased also one-third of the initial level in sliced carrots. Additional experiments were conducted to study the effect of irradiation and storage on the population of Listeria monocytogenes and Listeria innocua artificially inoculated on cut tomato and carrot. Cell numbers of both test organisms decreased by at least two log-cycles as an effect of 1 kGy dose. Our studies confirmed earlier findings on a temporary antilisterial effect of freshly cut carrot tissue. No re-growth of Listeria was observed during the studied storage period. The results of these studies suggest that irradiation with 1 kGy gamma rays could improve sufficiently the microbiological safety of the investigated pre-cut produce to satisfy the requirement of low microbial raw diets with acceptable nutritional quality and without diminishing significantly the organoleptic parameters of the

Susceptibility of wild carrot (Daucus carota ssp. carota) to Sclerotinia sclerotiorum

DEFF Research Database (Denmark)

Jensen, Brita Dahl; Finckh, M.R.; Munk, Lisa

2008-01-01

Sclerotinia soft rot, caused by Sclerotinia sclerotiorum, is a severe disease of cultivated carrots (Daucus carota ssp. sativus) in storage. It is not known whether Sclerotinia soft rot also affects wild carrots (D. carota ssp. carota), which hybridise and exchange genes, among them resistance...

The next generation of carotenoid studies in carrot (Daucus carota L.)

Science.gov (United States)

Orange carrot (Daucus carota L.) is one of the richest sources of naturally occurring ß-carotene while red and yellow carrot varieties contain large quantities of lycopene and lutein. The human body utilizes carotenoids, particularly ß-carotene (provitamin A) as a precursor for the production of ret...

Chemical and sensory quality of processed carrot puree as influenced by stress-induced phenolic compounds.

Science.gov (United States)

Talcott, S T; Howard, L R

1999-04-01

Physicochemical analysis of processed strained product was performed on 10 carrot genotypes grown in Texas (TX) and Georgia (GA). Carrots from GA experienced hail damage during growth, resulting in damage to their tops. Measurements included pH, moisture, soluble phenolics, total carotenoids, sugars, organic acids, and isocoumarin (6-MM). Sensory analysis was conducted using a trained panel to evaluate relationships between chemical and sensory attributes of the genotypes and in carrots spiked with increasing levels of 6-MM. Preharvest stress conditions in GA carrots seemed to elicit a phytoalexic response, producing compounds that impacted the perception of bitter and sour flavors. Spiking 6-MM into strained carrots demonstrated the role bitter compounds have in lowering sweetness scores while increasing the perception of sour flavor. Screening fresh carrots for the phytoalexin 6-MM has the potential to significantly improve the sensory quality of processed products.

Analysis of texture in baby carrot (Daucus carota) subjected to the process of ionizing radiation

International Nuclear Information System (INIS)

Nunes, Thaise C.F.; Rogovschi, Vladimir D.; Fabbri, Adriana D.T.; Sagretti, Juliana M.A.; Sabato, Susy F.

2011-01-01

The carrot is a vegetable of great economic value due to its versatility in the food industry and can be used as raw or minimally processed vegetable or aggregating value to the product, transforming the fresh carrots in baby carrots. It is well known that the application of gamma radiation in food may help in maintaining the quality of food. The aim of this study was to analyze the effects of the low doses of ionizing radiation on texture of minimally processed baby carrot after the processing in a Multipurpose 60 Co irradiator. It can be concluded that the treatment with low doses of gamma radiation keep the quality of fresh-cut baby carrot. (author)

A MYB transcription factor, DcMYB6, is involved in regulating anthocyanin biosynthesis in purple carrot taproots

OpenAIRE

Xu, Zhi-Sheng; Feng, Kai; Que, Feng; Wang, Feng; Xiong, Ai-Sheng

2017-01-01

Carrots are widely grown and enjoyed around the world. Purple carrots accumulate rich anthocyanins in the taproots, while orange, yellow, and red carrots accumulate rich carotenoids in the taproots. Our previous studies indicated that variation in the activity of regulatory genes may be responsible for variations in anthocyanin production among various carrot cultivars. In this study, an R2R3-type MYB gene, designated as DcMYB6, was isolated from a purple carrot cultivar. In a phylogenetic an...

7 CFR 319.56-43 - Baby corn and baby carrots from Zambia.

Science.gov (United States)

2010-01-01

... § 319.56-43 Baby corn and baby carrots from Zambia. (a) Immature, dehusked “baby” sweet corn (Zea mays L... consignments only. (b) Immature “baby” carrots (Daucus carota L. ssp. sativus) for consumption measuring 10 to...

Color changes kinetics during deep fat frying of carrot slice

Science.gov (United States)

Salehi, Fakhreddin

2018-05-01

Heat and mass transfer phenomena take place during frying cause physicochemical changes, which affect the colour and surface of the fried products. The effect of frying temperature on the colour changes and heat transfer during deep fat frying of carrot has been investigated. The colour scale parameters redness (a*), yellowness (b*) and lightness (L*), and color change intensity (ΔE) were used to estimate colour changes during frying as a function of oil temperature. L* value of fried carrot decreased during frying. The redness of fried carrot decreased during the early stages of frying, while it increased afterwards (become more red). A first-order kinetic equation was used for each one of the three colour parameters, in which the rate constant is a function of oil temperatures. The results showed that oil temperature has a significant effect on the colour parameters. Different kinetic models were used to fit the experimental data and the results revealed that the quadratic model was the most suitable to describe the color change intensity (ΔE) (R > 0.96). Center temperature of carrot slice increased with increase in oil temperature and time during frying.

Tunnel production enhances quality in organic carrot seed production

DEFF Research Database (Denmark)

Deleuran, L C; Boelt, B

2009-01-01

production of open-pollinated carrot varieties increased the yield and germination percentages when compared with normal field conditions. Yield was in the range of 100-250 g and 2-17 g seeds m-2 respectively, and germination percentage was 84-95 and 43-55, respectively. However, hybrid carrot seed...... production showed lower yields than did their open-pollinated counterparts. Yields ranging from 60-123 g seeds m-2 can be obtained, but the production needs to be carefully planned and monitored. Different growing systems in tunnels have been studied in both open-pollinated and hybrid carrot (Daucus carota L......In Denmark, organic vegetable seed production is possible for some of the late-maturing species when the maturing is performed in lightweight tunnels which are also relevant for the isolation of small-scale production. The tunnel system offers several advantages, e.g., it is possible to control...

Incidence and severity of cavity spot of carrot as affected by pigmentation, temperature, and rainfall

Science.gov (United States)

Field trials to determine the effect of carrot pigmentation and weather parameters on cavity spot of carrot (CS) were conducted in the Holland/Bradford Marsh region of Ontario between 2002 and 2009. Twenty three colored carrots from the USDA-ARS breeding program at the University of Wisconsin (5) an...

A MYB transcription factor, DcMYB6, is involved in regulating anthocyanin biosynthesis in purple carrot taproots.

Science.gov (United States)

Xu, Zhi-Sheng; Feng, Kai; Que, Feng; Wang, Feng; Xiong, Ai-Sheng

2017-03-27

Carrots are widely grown and enjoyed around the world. Purple carrots accumulate rich anthocyanins in the taproots, while orange, yellow, and red carrots accumulate rich carotenoids in the taproots. Our previous studies indicated that variation in the activity of regulatory genes may be responsible for variations in anthocyanin production among various carrot cultivars. In this study, an R2R3-type MYB gene, designated as DcMYB6, was isolated from a purple carrot cultivar. In a phylogenetic analysis, DcMYB6 was grouped into an anthocyanin biosynthesis-related MYB clade. Sequence analyses revealed that DcMYB6 contained the conserved bHLH-interaction motif and two atypical motifs of anthocyanin regulators. The expression pattern of DcMYB6 was correlated with anthocyanin production. DcMYB6 transcripts were detected at high levels in three purple carrot cultivars but at much lower levels in six non-purple carrot cultivars. Overexpression of DcMYB6 in Arabidopsis led to enhanced anthocyanin accumulation in both vegetative and reproductive tissues and upregulated transcript levels of all seven tested anthocyanin-related structural genes. Together, these results show that DcMYB6 is involved in regulating anthocyanin biosynthesis in purple carrots. Our results provide new insights into the regulation of anthocyanin synthesis in purple carrot cultivars.

Optimization of physicochemical and textural properties of pizza cheese fortified with soybean oil and carrot extract.

Science.gov (United States)

Motevalizadeh, Ehsan; Mortazavi, Seyed Ali; Milani, Elnaz; Hooshmand-Dalir, Moosa Al-Reza

2018-03-01

Response surface methodology (RSM) was used to optimize pizza cheese containing carrot extract. The effects of two important independent variables including soybean oil (5%-20%) and carrot extract (5%-20%) were studied on physicochemical and textural properties of pizza cheese containing carrot extract. According to the results, RSM was successfully used for optimizing formulation of pizza cheese containing carrot juice. Results of this study revealed that oil (A), carrot (B), AB, square term of carrot (B 2 ), B, AB, square term of oil (A 2 ), B 2 , AB, AB, A 2 B, A 2 , A 2 , A, A 2 , A 2 , AB, and AB 2 had the most effect on moisture, acidity, stretch, L*, a*, b*, hardness, meltability, springiness, peroxide value (PV), cohesiveness, chewiness, gumminess, fracture force, adhesiveness force, stiffness, flavor, and overall acceptability, respectively. A formulation upon 20% oil and 10.88% carrot extract was found as the optimal formulation for pizza cheese containing carrot extract. At the optimal formulation, PV, L*, a*, b*, meltability, stretch, cohesiveness, springiness, gumminess, chewiness, adhesive force, flavor, texture, and overall acceptability at the optimum formulation were measured 2.23, 82.51, -3.69, 18.05, 17.86, 85.61, 0.41, 7.874, 23.7, 0.27, 0.61, 3.50, 3.95, and 3.65, respectively.

Drinking carrot juice increases total antioxidant status and decreases lipid peroxidation in adults

Directory of Open Access Journals (Sweden)

Patil Bhimanagouda S

2011-09-01

Full Text Available Abstract Background High prevalence of obesity and cardiovascular disease is attributable to sedentary lifestyle and eating diets high in fat and refined carbohydrate while eating diets low in fruit and vegetables. Epidemiological studies have confirmed a strong association between eating diets rich in fruits and vegetables and cardiovascular health. The aim of this pilot study was to determine whether drinking fresh carrot juice influences antioxidant status and cardiovascular risk markers in subjects not modifying their eating habits. Methods An experiment was conducted to evaluate the effects of consuming 16 fl oz of daily freshly squeezed carrot juice for three months on cardiovascular risk markers, C-reactive protein, insulin, leptin, interleukin-1α, body fat percentage, body mass index (BMI, blood pressure, antioxidant status, and malondialdehyde production. Fasting blood samples were collected pre-test and 90 days afterward to conclude the study. Results Drinking carrot juice did not affect (P > 0.1 the plasma cholesterol, triglycerides, Apo A, Apo B, LDL, HDL, body fat percentage, insulin, leptin, interleukin-1α, or C-reactive protein. Drinking carrot juice decreased (P = 0.06 systolic pressure, but did not influence diastolic pressure. Drinking carrot juice significantly (P Conclusion Drinking carrot juice may protect the cardiovascular system by increasing total antioxidant status and by decreasing lipid peroxidation independent of any of the cardiovascular risk markers measured in the study.

Carotenoid crystal formation in Arabidopsis and carrot roots caused by increased phytoene synthase protein levels.

Directory of Open Access Journals (Sweden)

Dirk Maass

Full Text Available BACKGROUND: As the first pathway-specific enzyme in carotenoid biosynthesis, phytoene synthase (PSY is a prime regulatory target. This includes a number of biotechnological approaches that have successfully increased the carotenoid content in agronomically relevant non-green plant tissues through tissue-specific PSY overexpression. We investigated the differential effects of constitutive AtPSY overexpression in green and non-green cells of transgenic Arabidopsis lines. This revealed striking similarities to the situation found in orange carrot roots with respect to carotenoid amounts and sequestration mechanism. METHODOLOGY/PRINCIPAL FINDINGS: In Arabidopsis seedlings, carotenoid content remained unaffected by increased AtPSY levels although the protein was almost quantitatively imported into plastids, as shown by western blot analyses. In contrast, non-photosynthetic calli and roots overexpressing AtPSY accumulated carotenoids 10 and 100-fold above the corresponding wild-type tissues and contained 1800 and 500 microg carotenoids per g dry weight, respectively. This increase coincided with a change of the pattern of accumulated carotenoids, as xanthophylls decreased relative to beta-carotene and carotene intermediates accumulated. As shown by polarization microscopy, carotenoids were found deposited in crystals, similar to crystalline-type chromoplasts of non-green tissues present in several other taxa. In fact, orange-colored carrots showed a similar situation with increased PSY protein as well as carotenoid levels and accumulation patterns whereas wild white-rooted carrots were similar to Arabidopsis wild type roots in this respect. Initiation of carotenoid crystal formation by increased PSY protein amounts was further confirmed by overexpressing crtB, a bacterial PSY gene, in white carrots, resulting in increased carotenoid amounts deposited in crystals. CONCLUSIONS: The sequestration of carotenoids into crystals can be driven by the

New carrot and garlic germplasm to advance breeding and understand crop origins

Science.gov (United States)

The genetic variation provided by diverse plant germplasm is the basic building material used for crop improvement that shapes the crops we grow today. Wild carrot from the U.S. provided the cytoplasm used to develop a reliable system to produce hybrid carrots that account for most of the commercial...

Mass Proportion, Bioactive Compounds and Antioxidant Capacity of Carrot Peel as Affected by Various Solvents

Directory of Open Access Journals (Sweden)

Van Tang Nguyen

2016-11-01

Full Text Available The aim of this study was to determine the mass proportion of carrot root and the effects of four various solvents (methanol, water, ethanol and hexane on the contents of total phenolics and saponins as well as antioxidant capacity of carrot peel to identify an optimal solvent for effective extraction of bioactive compounds from carrot peel for further investigation. The results showed that carrot root consisted of body, heads and peel with their mass proportion of 83.19%, 5.01% and 14.19% by fresh weight, respectively. Among four solvents tested, methanol obtained the highest levels of extraction yield (54.02% by dry weight, total phenolic content (9.02 mg GAE/g dry weight and antioxidant capacity (DPPH radical scavenging capacity, cupric ion reducing antioxidant capacity, and ferric reducing antioxidant power from carrot peel, while water extracted the highest content of saponins (272.9 mg EE/g dry weight and possessed the maximum ABTS radical scavenging capacity. Therefore, methanol and water are considered for effective extraction of phenolics and saponins from carrot peel, respectively. The phenolic/saponin-enriched extracts are potential sources for further applications in the healthy food and/or pharmaceutical industries.

Hybrids between wild and cultivated carrots in Danish carrot fields

DEFF Research Database (Denmark)

Hauser, T.P.; Bjørn, G.K.

2001-01-01

, and as biennials they should normally not flower before harvest of the roots. Still, flowering individuals can be found in most Danish fields, and sometimes in very high numbers. At least 75% of the flowering plants are male fertile, with ca. 83% of the pollen being viable. More than half of the plants produce...... seeds. Pollen and seed dispersal from fields into wild carrot populations is probably rather frequent in Denmark. A closer inspection of the morphology of flowering plants indicate that some of these (2-60%) are bolters of pure cultivar origin, as indicated primarily by orange root colour. The remainder...

Carrot Juice Fermentations as Man-Made Microbial Ecosystems Dominated by Lactic Acid Bacteria.

Science.gov (United States)

Wuyts, Sander; Van Beeck, Wannes; Oerlemans, Eline F M; Wittouck, Stijn; Claes, Ingmar J J; De Boeck, Ilke; Weckx, Stefan; Lievens, Bart; De Vuyst, Luc; Lebeer, Sarah

2018-06-15

Spontaneous vegetable fermentations, with their rich flavors and postulated health benefits, are regaining popularity. However, their microbiology is still poorly understood, therefore raising concerns about food safety. In addition, such spontaneous fermentations form interesting cases of man-made microbial ecosystems. Here, samples from 38 carrot juice fermentations were collected through a citizen science initiative, in addition to three laboratory fermentations. Culturing showed that Enterobacteriaceae were outcompeted by lactic acid bacteria (LAB) between 3 and 13 days of fermentation. Metabolite-target analysis showed that lactic acid and mannitol were highly produced, as well as the biogenic amine cadaverine. High-throughput 16S rRNA gene sequencing revealed that mainly species of Leuconostoc and Lactobacillus (as identified by 8 and 20 amplicon sequence variants [ASVs], respectively) mediated the fermentations in subsequent order. The analyses at the DNA level still detected a high number of Enterobacteriaceae , but their relative abundance was low when RNA-based sequencing was performed to detect presumptive metabolically active bacterial cells. In addition, this method greatly reduced host read contamination. Phylogenetic placement indicated a high LAB diversity, with ASVs from nine different phylogenetic groups of the Lactobacillus genus complex. However, fermentation experiments with isolates showed that only strains belonging to the most prevalent phylogenetic groups preserved the fermentation dynamics. The carrot juice fermentation thus forms a robust man-made microbial ecosystem suitable for studies on LAB diversity and niche specificity. IMPORTANCE The usage of fermented food products by professional chefs is steadily growing worldwide. Meanwhile, this interest has also increased at the household level. However, many of these artisanal food products remain understudied. Here, an extensive microbial analysis was performed of spontaneous fermented

Rehydration kinetics of freeze-dried carrots

NARCIS (Netherlands)

Vergeldt, F.J.; Dalen, van G.; Duijster, A.J.; Voda, A.; Khalloufi, S.; Vliet, van L.J.; As, van H.; Duynhoven, van J.P.M.; Sman, van der R.G.M.

2014-01-01

Rehydration kinetics by two modes of imbibition is studied in pieces of freeze-dried winter carrot, after different thermal pre-treatments. Water ingress at room temperature is measured in real time by in situ MRI and NMR relaxometry. Blanched samples rehydrate substantially faster compared to

Development and sensory evaluation of yogurt added of “caviar” of carrot by children

Directory of Open Access Journals (Sweden)

Richtier Gonçalves Cruz

2015-12-01

Full Text Available This study aimed investigated to the preparation of yogurt with addition of “caviar” carrot and realize carried out sensory analysis of acceptance by children from elementary school. The “caviar” was prepared using the spherification technique with sodium alginate and added to the beaten-type yogurt without pulp addition at the ratio of one portion of the carrot for one portion of yogurt. Were conducted microbiological analyses and subsequently the sensory evaluation with children. The microbiological analyses showed that the product was fit for consumption. The average score in points obtained in the sensory analysis with 52 children was 6.7 (with 6.6 in males and 6.8 in females, it is that girls had higher scores (p < 0.05. The general index of acceptability to yogurt with “caviar” of carrot was 95.70%. It concludes that yogurt plus “caviar” carrot sensory was accepted because it presents high notes reflecting on their acceptability index, suggesting that this product is used as an alternative to increase the consumption of carrots by children.

Effect of foliar application of selenium on its uptake and speciation in carrot

DEFF Research Database (Denmark)

Kápolna, Emese; Hillestrøm, Peter René; Laursen, K.H.

2009-01-01

Carrot (Daucus carota) shoots were enriched by selenium using foliar application. Solutions of sodium selenite or sodium selenate at 10 and 100 mu g Se ml(-1), were sprayed on the carrot leaves and the selenium content and uptake rate of selenium were estimated by ICP-MS analysis. Anion and cation......(-1) (dry mass) in the carrot root whereas the selenium concentration in the controls was below the limit of detection at 0.045 mu g Se g(-1) (dry mass). Selenate-enriched carrot leaves accumulated as much as 80 mu g Se g(-1) (dry mass), while the selenite-enriched leaves contained approximately 50 mu...... g Se g(-1) (dry mass). The speciation analyses showed that inorganic selenium was present in both roots and leaves. The predominant metabolised organic forms of selenium in the roots were selenomethionine and gamma-glutamyl-selenomethyl-selenocysteine, regardless of which of the inorganic species...

Carrot (Daucus carota L . ssp. sativus (Hoffm. Arcang. as source of antioxidants

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Judita BYSTRICKÁ

2015-11-01

Full Text Available Carrot (Daucus carota L. s sp. sativus (Hoffm. Arcang. is a significant source of vitamins (A, B, C and beta carotene. Further it contains vitamins B, C, E, H, folic acid and pantothenic acid. Carrot is an important source of trace elements (K, Na, Ca, Mg, P, S, Mn, Fe, Cu and Zn. Consumption of carrot improves eyesight, lowers cholesterol and improves digestion. In this work we evaluated and compared content of total polyphenols, ß-caroteneand antioxidant activity in five varieties of carrot (’Jitka’, Kardila’, Katlen , Rubína’ and Koloseum Samples of carrot were collected at full maturity stages from area of Bardejov. Samples of fresh carrot were homogenized (25 g in 50 ml 80 % ethanol and analysed after sixteen hours. The content of the total polyphenols was determined by using the Folin-Ciocalteu reagent (FCR. The content of ß-carotene was determined spectrophotometricaly at 450 nm. Antioxidant activity was measured using a compound DPPH (2.2-diphenyl-1-picrylhydrazyl at 515.6 nm using spectrophotometer. Total polyphenols content in samples ranged from 81.25 ± 13.11 mg/kg to 113.69 ± 11.57 mg/kg and content of ß-carotenes ranged from 24.58 ± 2.38 mg/kg to 124.28 ± 3.54 mg/kg. We also evaluated and compared the antioxidant activity in selected varieties of ca rrot, which varied from 6.88 ± 0.92 % to 9.83 ± 0.62 %. Statistically si gnificant the highest value of total polyphenols was recorded in variety of Koloseum (113.69 ± 11.57 mg/kg. This variety is also character ized by the highest content of ß- carotene (124.28 ± 3.54 mg/kg as well as the highest value of antioxidant activity (9.83 ± 0.62 %.

High oxygen and high carbon dioxide modified atmospheres for shelf-life extension of minimally processed carrots

NARCIS (Netherlands)

Amanatidou, A.; Slump, R.A.; Gorris, L.G.M.; Smid, E.J.

2000-01-01

The impact of high O2 + high CO2 modified atmospheres (MA), on the preservation of minimally processed carrots was studied. A combination of 50% O2 + 30% CO2 prolonged the shelf life of sliced carrots compared to storage in air by 2 to 3 d. When the carrots received a pre-treatment with a 0.1%

Viability of L.casei in symbiotic carrot juice during fermentation and storage

OpenAIRE

Petreska Ivanovska, Tanja; Petrusevska Tozi, Lidija; Hadzieva, Jasmina; Smilkov, Katarina; Geskovski, Nikola; Mladenovska, Kristina

2011-01-01

Although dairy products are generally good matrices for the delivery of probiotics to humans and traditionally the most used, fruit juices are of growing interest, due to their pleasant taste profile and refreshing characteristics. However, the low survival rate of probiotics in fruit juices resulting from acid environment is of concern.In this study, carrot juice was inoculated with free probiotic cells of L. casei and symbiotic microparticles loaded with L. casei to compare the survival rat...

Relationship between carrot weevil infestation and parsley yield.

Science.gov (United States)

Torres, Angel N; Hoy, Casey W

2005-08-01

The relationship between numbers of carrot weevil, Listronotus oregonensis (LeConte), oviposition scars and parsley fresh weight and plant mortality was measured in research plots during 1999 and 2000. Fresh weight was measured in one to two cuttings of parsley planted on two planting dates. The average weight declined with increasing numbers of oviposition scars in the later planting in 1999. Compensatory growth in surviving plants may reduce this effect. Plant mortality increased as number of oviposition scars per plant increased in the second planting in both years and in the first cutting of the first planting in 2000. One oviposition scar per plant is sufficient to result in significant reduction in fresh weight per plant. In commercial parsley fields, the relationship between fresh weight of parsley per 30-cm row section of parsley was best described as a linear function of the proportion of plants with root feeding. Economic damage to parsley that is equivalent to the cost of controlling carrot weevil was estimated to result from approximately 1% of plants with root damage. Based upon this estimated economic injury level, we suggest an action threshold of 1% of plants containing carrot weevil oviposition scars earlier in the growing season when controls could be applied to prevent the damage.

Hybrids between cultivated and wild carrots in natural populations in Denmark

DEFF Research Database (Denmark)

Magnussen, L.S.; Hauser, Thure Pavlo

2007-01-01

Many cultivated plant species are able to hybridize with related wild plants. However, it is not clear whether their hybrids are able to survive and reproduce outside managed fields, and if cultivar genes introgress into wild populations. In areas where wild carrots co-occur with carrot root......-crops, pollen and seeds may flow from two different sources in the fields to the surrounding wild populations: from pure cultivar plants that occasionally flower, and from flowering 'bolters' that originate from hybridizations between wild (male) and cultivated carrots (female) in seed production fields...... by AFLP. Four hybrids were identified among the 71 plants analysed, and these were most likely F(2) or backcross individuals, sired by pollen from hybrid bolters. Wild populations close to fields were genetically somewhat more similar to cultivars than wild populations far from fields, suggesting...

Effect of soil compactness on the growth and quality of carrot

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Liisa Pietola

1995-05-01

Full Text Available Field experiments were performed in Southern Finland on three soil types: fine sand (1989-1991, clay (1989 and mull (1990-1991. The following soil mechanical treatments were applied to autumn ploughed land: soil loosening by ridge preparation (ridge distance 45 cm, rotary harrowing (to a depth of 20 cm, clay 15 cm, and soil compaction track by track by a tractor weighing 3 Mg (1 or 3 passes, wheel width 33 cm before seed bed preparation. One plot was untreated. These treatments were set up in April (on clay in May under moist soil conditions. Sprinkler irrigation (one application of 30 mm was applied to clay and fine sand when soil moisture in top soil had decreased to around 50% of plant-available water capacity. PVC cylinders (r = 15 cm, h = 60 cm were fixed in the experimental areas during the growing periods. At harvest, these cylinders were removed for specific analysis of tap and fibrous roots of carrot. Length and width of fibrous roots were quantified by image analysis in the USA. The impacts of soil loosening and partial compaction were determined by measuring soil physical parameters to a depth of 25 cm in mineral soils, and to greater depths in organic soil. Dry bulk densities of the plough layers increased with increasing tractor passes by 8%, 10% and 13% for fine sand, mull and clay soils, respectively. The lowest dry soil bulk density in the plough layer was obtained by rotary harrowing to a depth of 20 cm. Comparison of gamma ray transmission and gravimetric analysis indicated that dry soil bulk density was slightly lower when determined by gravimetric analysis. Increased soil bulk densities were reflected by increased water retention capacity (matric suction ≤ 10 kPa and greater penetrometer resistance. Relatively similar increases in bulk density increased the penetrometer resistance much less in mull than in fine sand. In contrast, greater bulk densities in the mull soil affected soil air composition adversely by decreasing

Insect Cell Culture

NARCIS (Netherlands)

Oers, van M.M.; Lynn, D.E.

2010-01-01

Insect cell cultures are widely used in studies on insect cell physiology, developmental biology and microbial pathology. In particular, insect cell culture is an indispensable tool for the study of insect viruses. The first continuously growing insect cell cultures were established from

Removal of heavy metals from aqueous solution by Carrot residues

International Nuclear Information System (INIS)

Eslamzadeh, T.; Nasernejad, B.; Bonakdar Pour, B.; Zamani, A.; Esmaail-Beygi, M.

2004-01-01

The removal of Copper(II), Zinc(II), and Chromium (III) from wastewater by carrot residues was investigated to evaluate cation exchange capacity. The effects of solution P H and co-ions were studied in batch experiments. Adsorption equilibria were initially rapidly established, and then decreased markedly after 10 min. Column experiments were carried out in a glass column filled with carrot residues to evaluate the metal removal capacity. The influences of the feed concentration and feed rate were also studied in order to compare the dynamic capacity for metal binding in different feed concentrations

Influence of gamma irradiation on phenolic compounds of minimally processed baby carrots

Energy Technology Data Exchange (ETDEWEB)

Hirashima, Fabiana K.; Fabbri, Adriana D.T.; Sagretti, Juliana M.A.; Nunes, Thaise C.F.; Sabato, Suzy F., E-mail: fmayumi@usp.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Galvao, Natascha S.; Lanfer-Marquez, Ursula M., E-mail: lanferum@usp.br [Universidade de Sao Paulo (FCF/USP), SP (Brazil). Faculdade de Ciencias Farmaceuticas

2013-07-01

Consumption of fresh fruits and vegetables provide several health benefits including risk reduction of oxidative stress-related diseases. These benefits have been associated with bioactive compounds, mainly phenolic compounds. Minimally processed products are a growing segment in food retail establishments due its practicality and convenience without significantly altering fresh-like characteristics. To extend the shelf life of these products, an application of ionizing radiation is an alternative, based on a physical and non-thermal method of preservation. The effect of irradiation on phenolic compounds of minimally processed baby carrots have not been reported in literature yet. The aim of this study was to evaluate the levels of phenolic compounds in baby carrots after the irradiation process. Samples of minimally processed baby carrots were purchased at a local supermarket and irradiated with doses of 0.5 and 1.0 kGy. Phenolic compounds were extracted from shredded carrots with MeOH and analyzed spectrophotometrically by the Folin Ciocalteau method using a gallic acid standard curve. The results showed that the phenolic contents decreased significantly (p<0.05) with increasing radiation dose. In non-irradiated baby carrots (control), the levels of phenolic compounds were about 330 μg eq. gallic acid/g, while irradiated samples with 0.5 kGy, showed an approximately 10% reduction when compared with the control. An irradiation dose of 1.0 kGy caused a loss of 20%. Although the radiation has affected the phenolic content, the process seems to be interesting by maintaining their fresh-like characteristics. (author)

Influence of gamma irradiation on phenolic compounds of minimally processed baby carrots

International Nuclear Information System (INIS)

Hirashima, Fabiana K.; Fabbri, Adriana D.T.; Sagretti, Juliana M.A.; Nunes, Thaise C.F.; Sabato, Suzy F.; Galvao, Natascha S.; Lanfer-Marquez, Ursula M.

2013-01-01

Consumption of fresh fruits and vegetables provide several health benefits including risk reduction of oxidative stress-related diseases. These benefits have been associated with bioactive compounds, mainly phenolic compounds. Minimally processed products are a growing segment in food retail establishments due its practicality and convenience without significantly altering fresh-like characteristics. To extend the shelf life of these products, an application of ionizing radiation is an alternative, based on a physical and non-thermal method of preservation. The effect of irradiation on phenolic compounds of minimally processed baby carrots have not been reported in literature yet. The aim of this study was to evaluate the levels of phenolic compounds in baby carrots after the irradiation process. Samples of minimally processed baby carrots were purchased at a local supermarket and irradiated with doses of 0.5 and 1.0 kGy. Phenolic compounds were extracted from shredded carrots with MeOH and analyzed spectrophotometrically by the Folin Ciocalteau method using a gallic acid standard curve. The results showed that the phenolic contents decreased significantly (p<0.05) with increasing radiation dose. In non-irradiated baby carrots (control), the levels of phenolic compounds were about 330 μg eq. gallic acid/g, while irradiated samples with 0.5 kGy, showed an approximately 10% reduction when compared with the control. An irradiation dose of 1.0 kGy caused a loss of 20%. Although the radiation has affected the phenolic content, the process seems to be interesting by maintaining their fresh-like characteristics. (author)

Enhanced accumulation of phytosterols and phenolic compounds in cyclodextrin-elicited cell suspension culture of Daucus carota.

Science.gov (United States)

Miras-Moreno, Begoña; Almagro, Lorena; Pedreño, M A; Sabater-Jara, Ana Belén

2016-09-01

In this work, suspension-cultured cells of Daucus carota were used to evaluate the effect of β-cyclodextrins on the production of isoprenoid and phenolic compounds. The results showed that the phytosterols and phenolic compounds were accumulated in the extracellular medium (15100μgL(-1) and 477.46μgL(-1), respectively) in the presence of cyclodextrins. Unlike the phytosterol and phenolic compound content, β-carotene (1138.03μgL(-1)), lutein (25949.54μgL(-1)) and α-tocopherol (8063.82μgL(-1)) chlorophyll a (1625.13μgL(-1)) and b (9.958 (9958.33μgL(-1)) were mainly accumulated inside the cells. Therefore, cyclodextrins were able to induce the cytosolic mevalonate pathway, increasing the biosynthesis of phytosterols and phenolic compounds, and accumulate them outside the cells. However, in the absence of these cyclic oligosaccharidic elicitors, carrot cells mainly accumulated carotenoids through the methylerythritol 4-phosphate pathway. Therefore, the use of cyclodextrins would allow the extracellular accumulation of both phytosterols and phenolic compounds by diverting the carbon flux towards the cytosolic mevalonate/phenylpropanoid pathway. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Development of Carrot Medium Suitable for Conidia Production of Venturia nashicola

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Eu Ddeum Choi

2017-02-01

Full Text Available The causal fungus of pear scab, Venturia nashicola, grows slowly and rarely produces conidia on artificial media in the laboratory, but it produced conidia on the Cheongah medium containing Cheongah powder. V. nashicola grew too slow to produce conidia until 15 days after cultivation but produced conidia with 4 × 10⁴ conidia/plate 30 days after cultivation on the Cheongah medium containing 1% Cheongah powder. V. nashicola showed a peak production of conidia with 4.5 × 10⁵ conidia/plate 60 days after cultivation on the carrot medium containing 2% carrot powder, one of the constituents of Cheongah powder. The carrot medium is considered to be the best medium to obtain conidia of V. nashicola in the laboratory until now. This is the first report on the development of a suitable medium for conidia production of V. nashicola, as far as we know.

Development of composition and technology of dairy dessert with carrot fiber

Directory of Open Access Journals (Sweden)

M. S. Belozerova

2016-01-01

Full Text Available The development of modern traditional technologies of processing of plant raw material leads to a reduction of content in it of fiber. In this regard, it becomes necessary to create a category of products, which are additionally introduced functional ingredients. Conducted researches on studying possibility of using the carrot fiber in the production of dairy products in order to enrich the composition of them dietary fiber. At the initial stage of the study determined the effect of temperature and time of exposure to the swelling capacity of carrot fiber. Based on the organoleptic evaluation of various samples of dairy products with food fibers sour cream with fat mass fraction 20 and 25% is offered as a dairy basis for production of a dessert. The technique of introducing carrot fiber in a product is perfected and its optimal dose equal to 3% is selected. As a flavor filler used cocoa syrup, which is best combined with sour cream base. The research carried out to determine the ratio of cocoa powder and sugar in cocoa syrup, which will allow to obtain a product with a harmonious combination of the studied components. Flavoring fillers are chosen and the rational dose equal to 30% by weight of the product allowing to receive a dessert with high consumer properties is defined. It was found that it is expedient to introduce carrot fiber into the mixture to prepare syrup, because technological parameters of these transactions are almost similar. At the final stage of the study examined the effect of carrot fiber for a period of validity of the developed product. It was found that shelf life of developed product is not reduced compared with the control sample containing no dietary fibers.

Drying of carrot slices in a triple pass solar dryer

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Seshachalam Kesavan

2017-01-01

Full Text Available An indirect triple pass forced convection solar dryer was developed and its performance was evaluated for drying of carrot slices. The drying experiments were carried out under the meteorological conditions of Coimbatore city in India during the year 2016. The experimental set-up consists of a blower, triple pass packed bed air collector (using sand with wire mesh absorber plate, and a drying chamber. The air mass flow rate was optimized to 0.062 kg/s. The initial moisture content of the carrot slices was reduced from 87.5% (on wet basis to the final moisture content of 10% (wet basis in 6 h duration. The thin layer drying characteristics were analyzed using twelve mathematical models available in open literature. The results showed that the pick-up efficiency of the dryer was varied in the range between 14 and 43% with an average air collector thermal efficiency of 44% during the experimentation. The drying characteristics of carrot slices was predicted with good degree of accuracy using Wang and Singh drying model.

Interaction of amatoxins with plant cells and RNA polymerases II: selection of amanitin-resistant cell lines and synthesis of amanitin-based affinity ligands

International Nuclear Information System (INIS)

Little, M.C.

1984-01-01

A series of experiments directed toward deriving basic information regarding plant RNA polymerase II is presented. The experiments described relate to the potential of isolating RNA polymerase II mutants in plants, using carrot cell cultures as models. Additionally, the synthesis of amanitin-based affinity ligands to immobilize isolated plant RNA polymerase II and associated transcriptional complexes is described. RNA polymerase II activities have been isolated from suspension cultures of carrot and compared to other plant RNA polymerases II with respect to subunit analysis and inhibition with α-amanitin. RNA polymerase II purified by polymin P absorption, DE52, phosphocellulose, and RNA-agarose chromatography is shown to copurify with proteins of 175 (and 200), 135, 70, 43, 28, 22, and 17 kdaltons apparent molecular weights. Conditions for accurate determination of amanitin inhibition of the enzyme are established using 3 H-amanitin and are presented for the first time for plant RNA polymerase II; RNA polymerase II from these cultures is shown to be inhibited by 50% at 3-5 nM by α-amanitin, a value 10-50 times lower than previously reported

Effectiveness of some substances in the control of carrot and parsley roots against fungal diseases.

Science.gov (United States)

Nawrocki, Jacek

2007-01-01

Field experiments were carried out in the years 2005 and 2006 on carrot cv. 'Koral' and 'Perfekcja', and parsley cv. 'Berlinska' and 'Cukrowa'. Effectiveness of substances: Biochikol 020 PC (biologically active substances BAS--chitosan 20 g/dm3), Bioczos BR (extract of garlic 10 g/1 brick) and Biosept 33 SL (extract of grapefruit 33%) on seedling roots of carrot and parsley was studied. As the standard fungicide Zaprawa Funaben T (carbendazim 20% + tiuram 45%) was used. Roots of carrot and parsley were treated one of tested substances spring immediately before planting seedling roots. During vegetation period the growth of seedling shoots and setting of seeds, and their infestation by fungal and bacterial pathogens was noticed. Among substances used for spring dressing of carrot and also parsley seedling roots, the best efficacy exhibited Zaprawa Funaben T in both years of observation. The highest yield of carrot seeds had combination roots cv. 'Koral' and parsley seeds roots cvs 'Berlińska' and 'Cukrowa' dressed Zaprawa Funaben T. Effectiveness of biopreparates Biochikol and Biosept was lower in comparison with the standard fungicide, but their protective effect was significantly higher than in control. Bioczos had the lowest control efficacy.

Effect of cover crops on emergence and growth of carrot (Daucus carota L. in no-plow and traditional tillage

Directory of Open Access Journals (Sweden)

Marzena Błażewicz-Woźniak

2015-03-01

Full Text Available The aim of the experiment was to determine the influence of cover crop biomass incorporated into the soil at different times and using different treatments on carrot emergence and growth. 7 species of cover crops were included in the study: Secale cereale, Avena sativa, Vicia sativa, Sinapis alba, Phacelia tanacetifolia, Fagopyrum esculentum, and Helianthus annuus.  Number of emerged carrot plants significantly depended on the cover crop used and on the method of pre-winter and spring pre-sowing tillage. Carrot emerged best after a rye or oats cover crop. Regardless of the cover crop species used, the largest number of carrots emerged in cultivation on ridges. In other variants of no-plow tillage, number of seedlings was significantly lower and did not differ from that under traditional plow tillage. The highest leaf rosettes were formed by carrot growing after a rye or oats cover crop. The highest rosettes were produced by carrots in the treatments where tillage was limited to the use of a tillage implement in spring and the lowest ones after pre-winter plowing. The effect of tillage on the emergence and height of carrot leaves largely depended on weather conditions in the successive years of the study. The largest number of leaves was found in carrots grown after a buckwheat cover crop and in cultivation without cover crop, while the smallest one after phacelia and white mustard. Carrots produced the largest number of leaves after a sunflower cover crop and the use of a tillage implement in spring, while the number of leaves was lowest when the mustard biomass was incorporated into the soil in spring. The use of cover crops significantly increased the mass of leaves produced by carrot as compared to the cultivation without cover crop. The largest mass of leaves was produced by carrots grown after the phacelia and mustard cover crops. Conventional plow tillage and pre-winter tillage using a stubble cultivator promoted an increase in the mass

Development of Blueberry and Carrot Juice Blend Fermented by Lactobacillus reuteri LR92

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Carolina Saori Ishii Mauro

2016-12-01

Full Text Available This study aimed to evaluate the blueberry and carrot juice blend as a fermentable substrate for Lactobacillus reuteri LR92, in order to develop a fermented non-dairy functional beverage. Analysis of cell viability, pH, and acidity were performed during the fermentation process. The resistance of the microorganism in the blend, under simulated gastrointestinal conditions and in storage at 4 °C for 28 days, was evaluated at the same time as the antioxidant potential of the fermented juice. After 40 h of fermentation, the L. reuteri population presented a logarithmic growth of three cycles, reaching count records of 10.26 ± 0.23 log CFU/mL and after 28 days of storage at 4 °C, the bacterial population maintained elevated numbers of viable cell (8.96 ± 0.08 log CFU/mL, with increase in the antioxidant capacity of the fermented blend. However, in the test of gastric simulation, the L. reuteri population had a logarithmic reduction of five cycles. In the presence of bile salts, the viability was maintained even after 150 min of incubation. This way, the results suggested that the blueberry and carrot blend juice can be considered as a good medium for the growth of L. reuteri, providing microbiological stability during refrigerated storage with elevated antioxidant capacity, which allows for the development of a non-dairy probiotic beverage.

Integration of cooking and vacuum cooling of carrots in a same vessel

Directory of Open Access Journals (Sweden)

Luiz Gustavo Gonçalves Rodrigues

2012-03-01

Full Text Available Cooked vegetables are commonly used in the preparation of ready-to-eat foods. The integration of cooking and cooling of carrots and vacuum cooling in a single vessel is described in this paper. The combination of different methods of cooking and vacuum cooling was investigated. Integrated processes of cooking and vacuum cooling in a same vessel enabled obtaining cooked and cooled carrots at the final temperature of 10 ºC, which is adequate for preparing ready-to-eat foods safely. When cooking and cooling steps were performed with the samples immersed in boiling water, the effective weight loss was approximately 3.6%. When the cooking step was performed with the samples in boiling water or steamed, and the vacuum cooling was applied after draining the boiling water, water loss ranged between 15 and 20%, which caused changes in the product texture. This problem can be solved with rehydration using a small amount of sterile cold water. The instrumental textural properties of carrots samples rehydrated at both vacuum and atmospheric conditions were very similar. Therefore, the integrated process of cooking and vacuum cooling of carrots in a single vessel is a feasible alternative for processing such kind of foods.

Greenhouse gas emissions in the life-cycle of carrots and tomatoes. Methods, data and results from a study of the types and amounts of carrots and tomatoes consumed in Sweden. With arable land use

Energy Technology Data Exchange (ETDEWEB)

Carlsson Kanyama, Annika

1997-03-01

Methods, data and results from an assessment of the arable land use and some greenhouse gas emissions during part of the life-cycle of the consumed carrots and tomatoes in Sweden during 1992-1993 are presented in the report. The life-cycle was delineated to transportation, storage, farm production and production of fertilizers. Carrots from six countries and tomatoes from four countries were analyzed. The study is reported with full transparency. The results are presented as the use of arable land (in m{sup 2}) and as the amount of greenhouse gas emissions (in g CO{sub 2} equivalents) required to sustain current Swedish consumption patterns of carrots and tomatoes. Emissions per kg of consumed tomato were 10 times higher than for carrots. Emissions from carrots were lowest when they were produced within or close to Sweden, while the opposite was the case for tomatoes. The key issues in the life-cycle of the analyzed carrots and tomatoes are identified and discussed. The general conclusions are that storage may be a key issue for vegetables with a long durability and adapted to a northern European climate. For vegetables with short durability, the key issue may be transportation if they are not adapted to a northern European climate but still cultivated in the open. The key issue for vegetables with a short durability may be energy requirements during farm production if the vegetables are not adapted to a northern European climate and therefore cultivated under glass. 3 figs, 57 tabs

Time-temperature dependent variations in beta-carotene contents in carrot using different spectrophotometric techniques

Science.gov (United States)

Ullah, Rahat; Khan, Saranjam; Shah, Attaullah; Ali, Hina; Bilal, Muhammad

2018-05-01

The current study presents time dependent variations in the concentration of beta-carotene in carrot under different storage-temperature conditions using UV–VIS and Raman spectrophotometric techniques. The UV–VIS absorption spectra of beta-carotene extracted from carrot shows three distinct absorption peaks at 442, 467, and 500 nm with maximum absorption at 467 nm. These absorption peaks are very much reproducible and are assigned to β-carotene. Similarly, Raman spectra of carrot samples also confirmed the three main Raman peaks of beta-carotene at shift positions 1003, 1150, and 1515 cm‑1. An overall decrease in beta-carotene content has been observed for time-temperature conditions. These results depict a decrease of about 40% in the content of beta-carotene when carrot samples were stored in a refrigerator (4 °C) for the first 20 d, whereas a decrease of about 25% was observed when carrot samples were stored in a freezer (‑16 °C) for the same period. The objective of this study is to investigate the possible use of Raman spectroscopy and UV–VIS spectroscopy for quick and detailed analysis of changes (degradation) in beta-carotene content associated with time and temperature in storage (frozen foods) in order to promote quality foods for consumers. Future study with a greater focus on the concentration/content of beta-carotene in other fruits/vegetables is also desirable.

Effect of Irradiation on the Hygienic, Chemical and Sensory Quality of Minimally Processed Fresh-Cut Carrot

International Nuclear Information System (INIS)

Mansour, F.A.; Hammad, A.A.; SwaiIam, H.M.; Abu EI-Nour, S.A.; Serag, M.S.

2008-01-01

The Average total bacteria count (TAPC) in the collected 15 samples of fresh-cut carrot ranged from 1.4 x 10 4 to 2.4x10 6 cfu/ g; lactic acid bacteria (LAB) between 5.0xl0 2 and 2.4xl0 5 cfu/ g; total mould and yeast (TM and Y) ranged from 2 cfu/ g indicating high level of microbial load and most of these samples were unacceptable from the view point of microbial load. All examined fresh-cut carrot samples had coliforms at value ranged from 43 to 1100 MPN/g and contained Escherichia coli in the range of 9 to 460 MPN/ g indicating that most of these samples were unsatisfactory due to the presence of E. coli at level higher than 100 MPN/ g. Enterococcus faecalis count was below the detectable level (100 cfu/ g). Staphylococcus aureus was detected in the majority (80 %) of the tested fresh-cut carrot samples, and were present at 1.0x10 2 to 2.3x10 3 cfu/g. Aeromonas hydrophila was detected in only 5 (33.3) samples with average counts 1.0x10 2 to 4.5xl0 2 cfu/g. The presence of E. coli, Staph. aureus and A. hydrophila in some samples of fresh-cut carrots is to be viewed as a health hazard. On the other hand, no Listeria monocytogenes or Salmonella spp. was detected in any of the fifteen fresh-cut carrot samples. Irradiation dose of 2 kGy extended the refrigeration shelf life of fresh-cut carrots to almost 12 days. The shelf-life for unirradiated samples was about 6 days. Irradiation dose of 4 kGy was identified to be the optimum irradiation dose for irradiating fresh-cut carrots. This irradiation dose greatly reduced the T APC (by 99.95 %) and decreased LAB and TM and Y to below the detectable level (<10 cfu/ g). Fresh-cut carrot samples receiving 4 kGy irradiation doses were free from studied indicator microorganisms and from pathogens via

Potato carrot agar with manganese as an isolation medium for Alternaria, Epicoccum and Phoma

DEFF Research Database (Denmark)

Sørensen, Jens Laurids; Mogensen, Jesper Mølgaard; Thrane, Ulf

2009-01-01

A semi-selective medium for isolation of Alternaria spp., Epicoccum sp. and Phoma spp. from soil and plant samples was developed. The basal medium was a modified potato carrot agar (PCA), containing 10 g/L of potato and carrot. It is known that the target genera sporulate well on standard PCA when...

A comprehensive approach to evaluate the freshness of strawberries and carrots

DEFF Research Database (Denmark)

Peneau, S.; Brockhoff, Per B.; Escher, F.

2007-01-01

of strawberries and carrots that varied in cultivar, as well as with time and conditions of storage. Product characteristics measured by descriptive sensory and physico-chemical analyses were related to consumer and expert panel (individual and consensus) ratings of freshness. Results showed that a large number...... of attributes contributed to the freshness of strawberries and carrots and that those were also indicators of the physiological ageing of these products. Our results suggest that consumer evaluation of fruit and vegetable freshness corresponds to an evaluation of this ageing process through the observation...

[Chemical, chemosensory and human-sensory experiments on taste and flavor of carrots].

Science.gov (United States)

Schaller, R G; Broda, S; Schnitzler, W H

1998-12-01

The relationship between sensory quality of carrots and their contents and composition of essential oils and total sugars as influenced by nitrogen fertilization was investigated. Carrots (Daucus carota L.) of the variety 'Nanthya' F1 (S&G Sandoz Seeds) were grown in Weihenstephan 1996 with three levels of inorganic nitrogen fertilization (3 levels in 4 replications). Medium- and higher-boiling flavour-components were extracted as essential oils and separated gas-chromatographically (GC-FID). Lower-boiling flavour-components were taken from the headspace and analysed chemosensorially. The human sensory assessments were performed by an untrained panel of 300 people (students and employees of the TU München)--these results were compared with those of the chemical analyses. Carrots with lower nitrogen application were found to taste more intensive, more fruity, sweeter and better and at the same time less bitter and less earthy. They had higher contents of total sugar and a higher percentage of dry matter. Fertilization with nitrogen does not only affect the quantity but also the composition of the essential oils. The taste intensive was positively correlated with the quantity of essential oils, the taste sweet was positively correlated with the content of total sugars. It was possible to differentiate the carrots from each other by chemo-sensorial headspace analyses according to their N-fertilization levels.

Production and characterization of curcumin microcrystals and evaluation of the antimicrobial and sensory aspects in minimally processed carrots.

Science.gov (United States)

Silva, Anderson Clayton da; Santos, Priscila Dayane de Freitas; Palazzi, Nicole Campezato; Leimann, Fernanda Vitória; Fuchs, Renata Hernandez Barros; Bracht, Lívia; Gonçalves, Odinei Hess

2017-05-24

Nontoxic conserving agents are in demand by the food industry due to consumers concern about synthetic conservatives, especially in minimally processed food. The antimicrobial activity of curcumin, a natural phenolic compound, has been extensively investigated but hydrophobicity is an issue when applying curcumin to foodstuff. The objective of this work was to evaluate curcumin microcrystals as an antimicrobial agent in minimally processed carrots. The antimicrobial activity of curcumin microcrystals was evaluated in vitro against Gram-positive (Bacillus cereus and Staphylococcus aureus) and Gram-negative (Escherichia coli and Pseudomonas aeruginosa) microorganisms, showing a statistically significant (p minimally processed carrots. Sensory analyses were carried out showing no significant difference (p minimally processed carrots without causing noticeable differences that could be detected by the consumer. One may conclude that the analyses of the minimally processed carrots demonstrated that curcumin microcrystals are a suitable natural compound to inhibit the natural microbiota of carrots from a statistical point of view.

Sand to Root Transfer of PAHs and PCBs by Carrots Grown on Sand with Pure Substances and Biosolids Amended Sand

OpenAIRE

Sablayrolles, Caroline; Montréjaud-Vignoles, Mireille; Silvestre, Jérôme; Patria, Lucie

2006-01-01

A study on behaviour of trace organic compounds (Polycyclic Aromatic Hydrocarbons, PAH, and Polychlorinated Biphenyls, PCB) in a sand-plant system has been carried out, with the reclamation of wastewater treatment plant biosolids for agriculture in mind. Carrot plants (Daucus carota) were grown on soilless culture (sand), to provide optimal transfer conditions, in plant containers inside a temperature regulated greenhouse. There were two types of experiment. The trace organic compounds have i...

ß-Carotene from Red Carrot Maintains Vitamin A Status, but Lycopene Bioavailability Is Lower Relative to Tomato Paste in Mongolian Gerbils

Science.gov (United States)

Red carrots contain lycopene in addition to ß-Carotene. The utility of red carrot as a functional food depends in part on the bioavailability of its constituent carotenoids. Lycopene bioavailability was compared in Mongolian gerbils (Meriones unguiculatus) fed freeze-dried red carrot and tomato pa...

Increasing cell culture population doublings for long-term growth of finite life span human cell cultures

Science.gov (United States)

Stampfer, Martha R; Garbe, James C

2015-02-24

Cell culture media formulations for culturing human epithelial cells are herein described. Also described are methods of increasing population doublings in a cell culture of finite life span human epithelial cells and prolonging the life span of human cell cultures. Using the cell culture media disclosed alone and in combination with addition to the cell culture of a compound associated with anti-stress activity achieves extended growth of pre-stasis cells and increased population doublings and life span in human epithelial cell cultures.

Comparison of purple carrot juice and β-carotene in a high-carbohydrate, high-fat diet-fed rat model of the metabolic syndrome.

Science.gov (United States)

Poudyal, Hemant; Panchal, Sunil; Brown, Lindsay

2010-11-01

Anthocyanins, phenolic acids and carotenoids are the predominant phytochemicals present in purple carrots. These phytochemicals could be useful in treatment of the metabolic syndrome since anthocyanins improve dyslipidaemia, glucose tolerance, hypertension and insulin resistance; the phenolic acids may also protect against CVD and β-carotene may protect against oxidative processes. In the present study, we have compared the ability of purple carrot juice and β-carotene to reverse the structural and functional changes in rats fed a high-carbohydrate, high-fat diet as a model of the metabolic syndrome induced by diet. Cardiac structure and function were defined by histology, echocardiography and in isolated hearts and blood vessels; liver structure and function, oxidative stress and inflammation were defined by histology and plasma markers. High-carbohydrate, high-fat diet-fed rats developed hypertension, cardiac fibrosis, increased cardiac stiffness, endothelial dysfunction, impaired glucose tolerance, increased abdominal fat deposition, altered plasma lipid profile, liver fibrosis and increased plasma liver enzymes together with increased plasma markers of oxidative stress and inflammation as well as increased inflammatory cell infiltration. Purple carrot juice attenuated or reversed all changes while β-carotene did not reduce oxidative stress, cardiac stiffness or hepatic fat deposition. As the juice itself contained low concentrations of carotenoids, it is likely that the anthocyanins are responsible for the antioxidant and anti-inflammatory properties of purple carrot juice to improve glucose tolerance as well as cardiovascular and hepatic structure and function.

Bioavailability and speciation of arsenic in carrots grown in contaminated soil

DEFF Research Database (Denmark)

Helgesen, H.; Larsen, Erik Huusfeldt

1998-01-01

increasing depression of growth with increasing level of contamination, At the experimental plots E-G with soil arsenic concentrations above 400 mu g g(-1) no carrots developed. Whether this effect was caused by arsenic or the concomitant copper content which ranged from 11 to 810 mu g g(-1) in the soil...... mixtures is unknown. The arsenic species extracted from the soils and carrots were separated and detected using anion-exchange HPLC coupled with ICP-MS, In the less contaminated soils from plots A and B arsenite (As-III) was more abundant than arsenate (As-V) in the soil using 1 mmole l(-1) calcium nitrate...

Development of Job’s tears ice cream recipes with carrot juice and pumpkin paste

OpenAIRE

Wiwat Wangcharoen

2011-01-01

Carrot juice and pumpkin paste were used as ingredients in Job’s tears ice cream. Carrot juice or pumpkin paste added at 50% was equally preferred by 100 consumers compared to the original Job’s tears ice cream. The new types of ice cream were lower in antioxidant capacity and higher in total phenolic content but could still be considered as potential antioxidant products. Purchase intent was significantly increased (p

Carrot Cultivar Evaluation: Soilless Media vs. Hydroponics

OpenAIRE

Pinnock, Derek R.; Bugbee, Bruce

2002-01-01

Nine cultivars of carrots were grown in a growth chamber. Each cultivar was grown both in hydroponic and soil-less media root-zone for sixty days. Three 30L tubs were used for each root-zone treatment. Three cultivars were planted in each tub, initially at 180 plants m-2 then thinned to 90 plants m-2 on day 45.

Construction and analysis of the transgenic carrot and celery plants expressing the recombinant thaumatin II protein

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Luchakivska Yu. S.

2015-08-01

Full Text Available Aim To obtain the transgenic carrot and celery plants able to express recombinant thaumatin II in order to increase plant stress tolerance. Methods. Agrobacterium-mediated transformation of the carrot and celery seedlings was used for obtaining the transgenic plants. Presence and transcription of the transgene in plant tissues were proved by PCR and RT-PCR analysis. The plants were tested for the biotic stress tolerance by in vitro antifungal and antibacterial activity assays and for the salinity and osmotic stress tolerance by plant survival test in presence of NaCl and PEG in different concentrations. Results. Transgenic plants able to express recombinant thaumatin II gene (transcription proved for 60–100 % were obtained by agrobacterial transformation. The transgenic carrot plant extracts inhibited the growth of the studied phytopathogenic bacteria strains but exhibited no antifungal activity. Survival level of transgenic plants under the salinity and osmotic stress effect was definitely higher comparing to the untransgenic ones. The analysis of the photosynthetic pigment content in the transgenic carrot plants showed no significant difference of this parameter under salinity stress that may indicate a possible protective activity of the recombinant protein. Conclusions. The obtained in our study transgenic carrot and celery plants able to express the recombinant thaumatin II gene were characterized by antibacterial activity and increased tolerance to salinity and osmotic stress factors.

The examination of parameters for lactic acid fermentation and nutritive value of fermented juice of beetroot, carrot and brewer’s yeast autolysate

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MILAN MAKSIMOVIC

2004-09-01

Full Text Available The conditions for lactic acid fermentation based on a mixture of beetoot juice (Beta vulgaris L. and carrot juice (Daucus carota L. and different content of brewer’s yeast autolysate with Lactobacillus plantarum A112 and with Lactobacillus acidophilus NCDO 1748 has been studied. Both cultures showed good biochemical activity in these mixtures. The production of lactic acid has been stimulated using a higher content of brewer’s yeast autolysate. In these mixtures, L. plantarum A112 showed better growth and lactic acid production than L. acidophilus NCDO 1748. From the data obtained through chemical analyses of the fermented products, it can be seen that the mixture of beetroot and carrot juice and brewer’s yeast autolysate is richer in minerals (Ca, P, Fe and b-carotene than fermented beetroot juice with the same content of brewer’s yeast autolysate.

Replacement Value of Untreated or Fungal Treated Carrot Leaves for Corn in Broiler Diet

International Nuclear Information System (INIS)

El-Faramawy, A.A.

2006-01-01

Three hundred (21 days old) Arbor Acre chicks were used to evaluate the replacement value of untreated or Aspergillus niger treated carrot leaves for corn in broiler diets. Birds were fed a control diet or diets in which 10% untreated or treated carrot leaves was quantitatively substituted for corn in the control diet. Replacement of yellow corn with 10% untreated and treated carrot leaves caused insignificant (P 0.01) in both experimental groups compared to control. These results denoted that although there were negligible changes in body weight and mortality rate in broiler chicken fed untreated and treated carrot leaves the amino acid profile of carcasses lessen their nutritive value which is in consequence reflected negatively on human amino acids intake. Poultry production represents one of the quickest means of correcting the anomaly of protein inadequacy, yet the rising cost of feed which represents 70-80% of the cost of production among other costs, is a major setback (Opera. 1996). The price of most conventional feed ingredients such as yellow corn, soybean meal and fish meal is so high in recent time that it is becoming uneconomical to use them in poultry feeding (Esonu et al., 2001). Consequently poultry feed researchers have been forced to seek alternative and cheaper feed resources

OPTIMIZATION OF PRETREATMENT CONDITIONS OF CARROTS TO MAXIMIZE JUICE RECOVERY BY RESPONSE SURFACE METHODOLOGY

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H. K. SHARMA

2006-12-01

Full Text Available Carrot juice was expressed in a hydraulic press using a wooden set up. Carrot samples pretreated at different designed combinations, using Central Composite Rotatable Design (CCRD, Response Surface Methodology (RSM, of pH, temperature and time were expressed and juice so obtained was characterized for various physico-chemical parameters which involved yield, TSS and water content, reducing sugars, total sugars and color (absorbance. The study indicated that carrots exposed to the different pretreatment conditions resulted in increased amount of yield than that of the control. The responses were optimized by numerical method and were found to be 78.23% yield, 0.93% color (abs, 3.41% reducing sugars, 5.53% total sugars, 6.69obrix, and 90.50% water content. All the derived mathematical models for the various responses were found to be fit significantly to predict the data.

Municipal Household Solid Waste Compost: Effects on Carrot ...

African Journals Online (AJOL)

An experiment was conducted to evaluate the impact of municipal household solid waste compost on N, P and K uptake and yield of carrot (Daucus carrota), using a coastal savanna Haplic Acrisol. Bulked samples of fresh solid waste from 45 households within the Cape Coast Municipality in the Central Region of Ghana ...

Functional gene polymorphism to reveal species history: the case of the CRTISO gene in cultivated carrots.

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Vanessa Soufflet-Freslon

Full Text Available Carrot is a vegetable cultivated worldwide for the consumption of its root. Historical data indicate that root colour has been differentially selected over time and according to geographical areas. Root pigmentation depends on the relative proportion of different carotenoids for the white, yellow, orange and red types but only internally for the purple one. The genetic control for root carotenoid content might be partially associated with carotenoid biosynthetic genes. Carotenoid isomerase (CRTISO has emerged as a regulatory step in the carotenoid biosynthesis pathway and could be a good candidate to show how a metabolic pathway gene reflects a species genetic history.In this study, the nucleotide polymorphism and the linkage disequilibrium among the complete CRTISO sequence, and the deviation from neutral expectation were analysed by considering population subdivision revealed with 17 microsatellite markers. A sample of 39 accessions, which represented different geographical origins and root colours, was used. Cultivated carrot was divided into two genetic groups: one from Middle East and Asia (Eastern group, and another one mainly from Europe (Western group. The Western and Eastern genetic groups were suggested to be differentially affected by selection: a signature of balancing selection was detected within the first group whereas the second one showed no selection. A focus on orange-rooted carrots revealed that cultivars cultivated in Asia were mainly assigned to the Western group but showed CRTISO haplotypes common to Eastern carrots.The carotenoid pathway CRTISO gene data proved to be complementary to neutral markers in order to bring critical insight in the cultivated carrot history. We confirmed the occurrence of two migration events since domestication. Our results showed a European background in material from Japan and Central Asia. While confirming the introduction of European carrots in Japanese resources, the history of Central Asia

Profiling of the Terpene Metabolome in Carrot Fruits of Wild ( Daucus carota L. ssp. carota) Accessions and Characterization of a Geraniol Synthase.

Science.gov (United States)

Yahyaa, Mosaab; Ibdah, Muhammad; Marzouk, Sally; Ibdah, Mwafaq

2018-03-14

Fruits from wild carrot ( Daucus carota L. ssp. carota) have been used for medicinal purposes since ancient times. The oil of its seeds, with their abundant monoterpenes and sesquiterpenes, has drawn attention in recent years because of its potential pharmaceutical application. A combined chemical, biochemical, and molecular study was conducted to evaluate the differential accumulation of terpene volatiles in carrot fruits of wild accessions. This work reports a similarity-based cloning strategy identification and functional characterization of one carrot monoterpene terpene synthase, WtDcTPS1. Recombinant WtDcTPS1 protein produces mainly geraniol, the predominant monoterpene in carrot seeds of wild accession 23727. The results suggest a role for the WtDcTPS1 gene in the biosynthesis of carrot fruit aroma and flavor compounds.

IP3 stimulates CA++ efflux from fusogenic carrot protoplasts

International Nuclear Information System (INIS)

Rincon, M.; Boss, W.F.

1986-01-01

Polyphosphoinositide breakdown plays an important role in signal transduction in animal cells (Berridge and Irvine, 1984, Nature, 312:315). Upon stimulation, phospholipase C hydrolyzes phosphatidylinositol 4,5-bisphosphate to inositol 1,4,5-trisphosphate (IP 3 ) and diacylglycerol both of which act as cellular second messengers. IP 3 mobilizes Ca ++ from internal stores, hence the cytosolic free Ca ++ concentration increases and those physiological activities regulated by Ca ++ are stimulated. To test if plant cells also responded to IP 3 , Ca ++ efflux studies were done with fusogenic carrot protoplasts released in EGTA. The protoplasts were preloaded with 45 Ca ++ placed in a Ca ++ -free medium, and efflux determined as 45 Ca ++ loss from the protoplasts. IP 3 (10-20μM) caused enhanced 45 Ca ++ efflux and the response was sustained for at least 15 min. In plants, as in animals, the observed IP 3 -enhanced 45 Ca ++ efflux suggested that IP 3 released Ca ++ from internal stores, and the increased free cytosolic Ca ++ activated Ca ++ pumping mechanisms which restored the Ca ++ concentration in the cytosol to the normal level

Rapid changes in plasma membrane protein phosphorylation during initiation of cell wall digestion

International Nuclear Information System (INIS)

Blowers, D.P.; Boss, W.F.; Trewavas, A.J.

1988-01-01

Plasma membrane vesicles from wild carrot cells grown in suspension culture were isolated by aqueous two-phase partitioning, and ATP-dependent phosphorylation was measured with [γ- 32 P]ATP in the presence and absence of calcium. Treatment of the carrot cells with the cell wall digestion enzymes, driselase, in a sorbitol osmoticum for 1.5 min altered the protein phosphorylation pattern compared to that of cells treated with sorbitol alone. Driselase treatment resulted in decreased phosphorylation of a band of M r 80,000 which showed almost complete calcium dependence in the osmoticum treated cells; decreased phosphorylation of a band of M r 15,000 which showed little calcium activation, and appearance of a new band of calcium-dependent phosphorylation at M r 22,000. However, protein phosphorylation was decreased. Adding driselase to the in vitro reaction mixture caused a general decrease in the membrane protein phosphorylation either in the presence or absence of calcium which did not mimic the in vivo response. Cells labeled in vivo with inorganic 32 P also showed a response to the Driselase treatment. An enzymically active driselas preparation was required for the observed responses

Impacts of irrigation regimes with saline water on carrot productivity and soil salinity

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Kamel Nagaz

2012-01-01

Full Text Available A three-year study was conducted to evaluate the effects of different irrigation regimes with saline water on soil salinity, yield and water productivity of carrot as a fall-winter crop under actual commercial-farming conditions in the arid region of Tunisia. Carrot was grown on a sandy soil and surface-irrigated with a water having an ECi of 3.6 dS/m. For the three years, a complete randomized block design with four replicates was used to evaluate five irrigation regimes. Four irrigation methods were based on the use of soil water balance (SWB to estimate irrigation amounts and timing while the fifth consisted of using traditional farmers practices. SWB methods consisted in replacement of cumulated ETc when readily available water is depleted with levels of 100% (FI-100, 80% (DI-80 and 60% (DI-60. FI-100 was considered as full irrigation while DI-80 and DI-60 were considered as deficit irrigation regimes. Regulated deficit irrigation regime where 40% reduction is applied only during ripening stage (FI-DI60 was also used. Farmer method (Farmer consisted in giving fixed amounts of water (25 mm every 7 days from planting till harvest. Results on carrot production and soil salinization are globally consistent between the three-year experiments and shows significant difference between irrigation regimes. Higher soil salinity in the root zone is observed at harvest under DI-60 (3.1, 3.4, 3.9 dS/m, respectively, for the three years and farmer irrigation (3.3, 3.6, 3.9 dS/m treatments compared to FI-100 treatment (2.3, 2.6 and 3.1 dS/m. Relatively low ECe values were also observed under FI-DI60 and DI-80 treatments with respectively (2.7, 3, 3.5 dS/m and (2.5, 2.9, 3.3 dS/m. ECe values under the different irrigation treatments were generally lower than or equal to the EC of irrigation water used. Rainfall received during fall and/or winter periods (57, 26 and 29 mm, respectively, during the three years contributed probably to leaching soluble

The Impact of Carrot Enriched in Iodine through Soil Fertilization on Iodine Concentration and Selected Biochemical Parameters in Wistar Rats

Science.gov (United States)

Piątkowska, Ewa; Kopeć, Aneta; Bieżanowska-Kopeć, Renata; Pysz, Mirosław; Kapusta-Duch, Joanna; Koronowicz, Aneta Agnieszka; Smoleń, Sylwester; Skoczylas, Łukasz; Ledwożyw-Smoleń, Iwona; Rakoczy, Roksana; Maślak, Edyta

2016-01-01

Iodine is one of the trace elements which are essential for mammalian life. The major objective of iodine biofortification of plants is to obtain food rich in this trace element, which may increase its consumption by various populations. Additionally, it may reduce the risk of iodine deficiency diseases. In this research for the first time we have assessed the bioavailability of iodine from raw or cooked carrot biofortified with this trace element on iodine concentration in selected tissues and various biochemical parameters as well as mRNA expression of some genes involved in iodine metabolism in Wistar rats. Statistically, a significantly higher iodine level was determined in urine, faeces and selected tissues of rats fed a diet containing biofortified raw carrot as compared to a diet without iodine and a diet containing control cooked carrot. Biofortified raw carrot significantly increased triiodothyronine concentration as compared to animals from other experimental groups. The highest thyroid stimulating hormone level was determined in rats fed control cooked carrots. mRNA expression of selected genes was affected by different dietary treatment in rats’ hearts. Biofortified raw and cooked carrot could be taken into account as a potential source of iodine in daily diets to prevent iodine deficiency in various populations. PMID:27043135

Effect of low doses of irradiation on the carotenoids in head to eat carrots

International Nuclear Information System (INIS)

Lima, K.S.C.; Lima, A.L.S.; Freitas, L.C.; Della-Modesta, R.C.; Godoy, R.L.O.

2004-01-01

This study aims was to evaluate the effect of low doses of g radiation on the total carotenoids, α and β-carotene content in minimally processed carrots, during the shelflife. Carrots are the mains vegetable source of carotenoids provitamin A (α and β-carotene). According to the Family Budget Survey (FBS) carried out in the Brazilian Southeast, within the roots and tubers group, carrots are widely consumed. The carotenoid stability varies largely during the stages of processing and storage, depending upon structure, temperature, oxygen availability, light exposure, humidity content, water activity and acid, metal anti-oxidant and pro-oxidant presence. The minimally processed carrots in this experiment were manually peeled, rinsed, cutted into diskis, packaged under 5% O 2 / 10% CO 2 and 21% O 2 (sintetic air), g ionizing radiation treatments was carried out with a 137 Cs source, of 0,25, 0,50, 0,75 and 1,0kGy doses, and shelf-stored at 5°C for 24 days. Total carotenoids quantification was by 449nm spectrophotometer. Determination of a and β-carotenes was made by High Performance Liquid Chromatography (HPLC). The different treatments and control group were, too, evaluated by analysing of colour and volatiles, by gas chromatography/mass spectroscopy with solid phase microextration (CG-MS/SPME), for study the significant carotenoids losses during the process [pt

Water properties and structure of pork sausages as affected by high-pressure processing and addition of carrot fibre

DEFF Research Database (Denmark)

Møller, Sandie Mejer; Grossi, Alberto Blak; Christensen, Mette

2011-01-01

The effects of high-pressure processing (HPP) and addition of carrot fibre on pork sausages have been studied using NMR T(2) relaxometry and measurements of water-binding capacity (WBC) by centrifugation. Significant effects of temperature (raw, 40, 50, or 60°C), holding time (1s, 3, 6, or 9min......), and addition of carrot fibre on the distribution and mobility of water were found. However, the effect of carrot fibre could not be explained by structural changes in the sausages when examined by confocal laser scanning microscopy (CLSM). Correlations between T(2) relaxation measurements and WBC determined...... by centrifugation revealed that T(2) relaxation times were able to explain more than 90% of the variation in WBC for both non-pressure and pressure-treated sausages. However, only 49% of the variation was explained for pressure-treated sausages with carrot fibre, indicating that combining addition of fibre and high...

Study on combined effects of acidification and sonication on selected quality attributes of carrot juice during storage

International Nuclear Information System (INIS)

Jabbar, S.; Hu, B.; Ali, S.

2014-01-01

This study evaluated the combined effects of acid blanching and sonication treatments on selected quality parameters of carrot juice stored at 4 degree C for 18 days. Carrots were blanched in acidified water (40g/L citric acid) at 100 degree C for 4 min and the juice was then extracted. Sonication of the juice was done at an amplitude level of 70% and a frequency of 20 kHz for 2 min at 15 degree C, keeping the pulse duration of 5 Sec on and 5 Sec off. As results, the combined treatment of acidification and sonication of carrot juice showed a significant decrease in pH and increase (P < 0.05) in acidity which remained stable during storage period. No significant changes were observed in Brix. Color values (L, a, b) and non enzymatic browning (NEB) influenced significantly in acidified and sonicated carrot juice during storage period. Maximum stability of total phenol, total antioxidant capacity, cloud value and ascorbic acid were also observed in the combined treatment of acidification and sonication. The findings of this study indicated that the combined treatments of acidification and sonication may successfully be utilized for the production of high quality carrot juice with improved stability of total phenol, total antioxidant capacity, cloud value and ascorbic acid during 18 days of storage. (author)

Effects of ethylene on gene expression in carrot roots

International Nuclear Information System (INIS)

Nichols, S.E.

1984-01-01

To investigate ethylene effects on expression of genetic information, cDNA clones corresponding to ethylene-induced carrot root mRNAs were constructed and isolated. RNA dot blot analysis showed that for the three clones studied peak cytosolic mRNA prevalence occurred at 21 hours of treatment followed thereafter by rapid messenger decay. DNA filter excess hybridization to in vitro synthesized nuclear RNA showed that the ethylene-induced mRNA increase is engendered by transcription of previously quiescent genes. The kinetics and magnitude of changes in mRNA prevalence parallel changes in transcriptional activity; therefore, the ethylene effect is primarily at the level of the transcription. In vivo pulse labelling with [ 35 S]-methionine showed that between 18 and 27 hours of ethylene treatment a 2.5 fold increase in translational efficiency occurred for one message studied. The resulting protein is the predominant protein synthesized in carrots treated with ethylene for 27 hours. Thus, ethylene exerts multiple regulatory controls on the expression of genetic information

Qualidade de produto minimamente processado à base de abóbora, cenoura, chuchu e mandioquinha-salsa Maintenance of the quality of fresh-cut products made up of pumpkin, carrot, chayote, and arracacha (peruvian carrot

Directory of Open Access Journals (Sweden)

Juliana Alvarenga Alves

2010-09-01

Full Text Available O objetivo deste trabalho foi avaliar a qualidade de produto minimamente processado, à base de quatro hortaliças - abóbora, cenoura, chuchu e mandioquinha-salsa, armazenado a 5 ºC por 8 dias. Observou-se que os teores de umidade, fibra, proteína, cinza e fração glicídica das quatro hortaliças não foram afetados pelo tempo de armazenamento, entretanto o teor de extrato etéreo aumentou. Durante o armazenamento, o teor de vitamina C e de acidez titulável diminuiu em todas as hortaliças. O teor de β-caroteno do chuchu não alterou, entretanto, aumentou na abóbora, na cenoura e na mandioquinha-salsa. O pH das quatro hortaliças aumentou com o armazenamento. Os teores de sólidos solúveis da cenoura e da mandioquinha-salsa aumentaram, não sendo afetados na abóbora e no chuchu. As notas de aparência do "mix" não foram inferiores a 7 (gostei moderadamente, durante o armazenamento. Os coliformes a 35 ºC presentes no "mix" aumentaram e não foi constatada a presença de coliformes a 45 ºC e Salmonella sp. em nenhum tempo avaliado. Conclui-se que a vida útil, entendida sob os aspectos nutricionais, sensoriais e microbiológicos, pode ser estabelecida em 8 dias sob refrigeração para abóbora, cenoura, chuchu e mandioquinha-salsa submetidos ao processamento mínimo.The goal of this study was to evaluate the quality of fresh-cut products made up of four vegetables: pumpkin, carrot, chayote, and arracacha (peruvian carrot stored at 5 ºC for 8 days. It was observed that the contents of humidity, fiber, protein, ash, and glucidic fraction of the four vegetables were not affected by the time of storage; however, the content of ethereal extract increased. During storage, the content of vitamin C and titratable acidity decreased in all the vegetables. The content of β-carotene of chayote did not change, whereas the content in the pumpkin, carrot, and the peruvian carrot increased. The pH of the four vegetables increased during storage

Uptake of polybrominated diphenyl ethers by carrot and lettuce crops grown in compost-amended soils.

Science.gov (United States)

Bizkarguenaga, E; Iparraguirre, A; Oliva, E; Quintana, J B; Rodil, R; Fernández, L A; Zuloaga, O; Prieto, A

2016-02-01

The uptake of polybrominated diphenyl ethers (PBDEs) by carrot and lettuce was investigated. Degradation of PBDEs in soil in the absence of the plants was discarded. Different carrot (Nantesa and Chantenay) and lettuce (Batavia Golden Spring and Summer Queen) varieties were grown in fortified or contaminated compost-amended soil mixtures under greenhouse conditions. After plant harvesting, roots (core and peel) and leaves were analyzed separately for carrot, while for lettuce, leaves and hearts were analyzed together. The corresponding bioconcentration factors (BCFs) were calculated. In carrots, a concentration gradient of 2,2',3,4,4',5'-hexabromodiphenyl ether (BDE-138) became evident that decreased from the root peel via root core to the leaves. For decabromodiphenyl ether (BDE-209) at the low concentration level (7 and 20 ng g(-1)), the leaves incorporated the highest concentration of the target substance. For lettuce, a decrease in the BCF value (from 0.24 to 0.02) was observed the higher the octanol-water partition coefficient, except in the case of BDE-183 (BCF = 0.51) and BDE-209 (BCF values from 0.41 to 0.74). Significant influence of the soils and crop varieties on the uptake could not be supported. Metabolic debromination, hydroxylation or methylation of the target PBDEs in the soil-plant system was not observed.

Detection and transmission of Carrot torrado virus, a novel putative member of the Torradovirus genus.

Science.gov (United States)

Rozado-Aguirre, Zuriñe; Adams, Ian; Collins, Larissa; Fox, Adrian; Dickinson, Matthew; Boonham, Neil

2016-09-01

A new Torradovirus tentatively named Carrot torrado virus (CaTV) was an incidental finding following a next generation sequencing study investigating internal vascular necrosis in carrot. The closest related viruses are Lettuce necrotic leaf curl virus (LNLCV) found in the Netherlands in 2011 and Motherwort yellow mottle virus (MYMoV) found in Korea in 2014. Primers for reverse transcriptase-PCR (RT-PCR) and RT-qPCR were designed with the aim of testing for the presence of virus in plant samples collected from the field. Both methods successfully amplified the target from infected samples but not from healthy control samples. The specificity of the CaTV assay was also checked against other known carrot viruses and no cross-reaction was seen. A comparative study between methods showed RT-qPCR was the most reliable method, giving positive results in samples where RT-PCR fails. Evaluation of the Ct values following RT-qPCR and a direct comparison demonstrated this was due to improved sensitivity. The previous published Torradovirus genus specific RT-PCR primers were tested and shown to detect CaTV. Also, virus transmission experiments carried out suggest that unlike other species of the same genus, Carrot torrado virus could be aphid-transmitted. Copyright © 2016 Elsevier B.V. All rights reserved.

The Rise of Carrots and the Decline of Sticks

NARCIS (Netherlands)

de Geest, G.; Dari-Mattiacci, G.

2013-01-01

There is a remarkable tendency in modern legal systems to increasingly use carrots. This trend is not limited to legal systems but can also be observed in, for instance, parenting styles, social control mechanisms, and even law schools’ teaching methods. Yet, at first glance, sticks appear to be a

The effect of preliminary processing and different methods of cooking on the iodine content and selected antioxidative properties of carrot (Daucus carota L. biofortified with (potassium iodine

Directory of Open Access Journals (Sweden)

Kapusta-Duch Joanna

2017-06-01

Full Text Available Carrot is a vegetable that contains many nutrients and has strong antioxidant activity as well as pro-health potential. The level of bioactive compounds is strongly connected with the production chain. The thermal treatment of food products induces several biological, physical and chemical changes. In this study, changes in the levels of iodine, total carotenoids, total polyphenols as well as the antioxidant activity of unpeeled and peeled controls and carrots biofortified with (potassium iodine (KJ during cultivation due to the cooking and steaming process were investigated. The use of thermal processes resulted in a lower concentration of iodine in the roots of the control as well as in carrots biofortified with (potassium iodine. In addition, peeling carrots caused higher losses of this trace element in the control and the biofortified carrots cooked or steamed for various times. In this study, a significant growth of the total carotenoids in peeled carrots biofortified with (potassium iodine and of the total polyphenols in unpeeled carrots biofortified with (potassium iodine under the influence of the cooking and steaming processes was observed compared with raw peeled and unpeeled biofortified carrots, respectively. Antioxidant activity significantly increased in the unpeeled and peeled carrots biofortified with (potassium iodine under all thermal treatments in comparison with the raw unpeeled and peeled biofortified carrots.

Effect of refrigerated storage on the probiotic survival and sensory properties of milk/carrot juice mix drink

OpenAIRE

Daneshi,Mohammad; Ehsani,Mohammad Reza; Razavi,Seyed Hadi; Labbafi,Mohsen

2013-01-01

Background: There is a genuine interest in the development of probiotic milk and juice based beverages because they are a good-vehicle to deliver probiotic microorganisms to consumers. For this purpose, the viability and metabolism of four probiotic strains (Lactobacillus acidophilus LA5, Bifidobacterium lactis BB12, L. rhamnosus and L. plantarum) were studied in non-fermented milk and carrot juice mix drink. The drinks were evaluated in 5 days interval for viable cell count, pH, acidity, sed...

Antioxidant Activity of Cabbage and/or Carrot Against Oxidative Stress Induced by Gamma Irradiation in Male Albino Rats

International Nuclear Information System (INIS)

Hamza, R.G.; Mahmoud, K.A.

2011-01-01

Several studies indicated that diets rich in fruits and vegetables are protective against diseases, and populations that consume such diets have higher plasma antioxidants and exhibit lower risk of cancer and cardiovascular diseases. Vegetable is considered major dietary source of fibers and antioxidants such as polyphenols, flavonoids and carotenoids that can protect against different dietary disorders. The present study was carried out to investigate the potential protective effects of cabbage and/or carrot against oxidative stress induced by gamma irradiation in male albino rats. Chemical composition and phenolic contents in cabbage and carrot were determined. Male albino rats were exposed to 5 Gy (single dose with rate 0.46 Gy/min) of whole body gamma irradiation. Thirty five rats were randomly divided into five groups as follow: group 1: control (rats fed on balanced diet for 6 weeks), group 2: irradiated (rats were exposed to whole gamma irradiation and fed on balanced diet for 6 weeks) and groups 3, 4 and 5: irradiated rats fed on balanced diet and received cabbage 15%, carrot 15% and a combination of cabbage and carrot, respectively. The results obtained revealed that the administration of cabbage and/or carrot diet significantly reduced the changes induced by gamma irradiation in the serum level of glucose and liver function parameters; serum aminotransferases (AST, ALT), alkaline phosphatase (ALP), total protein and albumin. In addition, significant improvements were observed in the serum levels of total cholesterol (TC), triglycerides (TG), low density lipoprotein-cholesterol (LDL-C) and high density lipoprotein-cholesterol (HDL-C). Significant enhancement in hepatic antioxidant enzymes; superoxide dismutase (SOD) and catalase (CAT), was observed. The levels of reduced glutathione (GSH) associated with remarkable decrease in the level of lipid peroxidation (TBARS) were observed. Accordingly, it could be concluded that consumption of cabbage and/or carrot

UVA, UVB and UVC Light Enhances the Biosynthesis of Phenolic Antioxidants in Fresh-Cut Carrot through a Synergistic Effect with Wounding

Directory of Open Access Journals (Sweden)

Bernadeth B. Surjadinata

2017-04-01

Full Text Available Previously, we found that phenolic content and antioxidant capacity (AOX in carrots increased with wounding intensity. It was also reported that UV radiation may trigger the phenylpropanoid metabolism in plant tissues. Here, we determined the combined effect of wounding intensity and UV radiation on phenolic compounds, AOX, and the phenylalanine ammonia-lyase (PAL activity of carrots. Accordingly, phenolic content, AOX, and PAL activity increased in cut carrots with the duration of UVC radiation, whereas whole carrots showed no increase. Carrot pies showed a higher increase compared to slices and shreds. Phenolics, AOX, and PAL activity also increased in cut carrots exposed to UVA or UVB. The major phenolics were chlorogenic acid and its isomers, ferulic acid, and isocoumarin. The type of UV radiation affected phenolic profiles. Chlorogenic acid was induced by all UV radiations but mostly by UVB and UVC, ferulic acid was induced by all UV lights to comparable levels, while isocoumarin and 4,5-diCQA was induced mainly by UVB and UVC compared to UVA. In general, total phenolics correlated linearly with AOX for all treatments. A reactive oxygen species (ROS mediated hypothetical mechanism explaining the synergistic effect of wounding and different UV radiation stresses on phenolics accumulation in plants is herein proposed.

Identification of volatile organic compounds (VOCs in different colour carrot (Daucus carota L. cultivars using static headspace/gas chromatography/mass spectrometry

Directory of Open Access Journals (Sweden)

Zehra Güler

2015-12-01

Full Text Available Volatile organic compounds (VOCs as well as sugar and acid contents affect carrot flavour. This study compared VOCs in 11 carrot cultivars. Gas chromatography/mass spectrometry using static headspace technique was applied to analyse the VOCs. The number of VOCs per sample ranged from 17 to 31. The primarily VOCs identified in raw carrots with the exception of “Yellow Stone” were terpenes, ranging from 65 to 95%. The monoterpenes with values ranging from 31 to 89% were higher than those (from 2 to 15% of sesquiterpenes. Monoterpene α-terpinolene (with ranging from 23 to 63% and (--α-pinene (26%, and alcohol ethanol (35% was the main VOC in extracts from the nine carrot cultivars, “Purple” and “Yellow Stone”, respectively. As a result, among 16 identified monoterpenes, 7 monoterpenes (--α-pinene, (--β-pinene, β-myrcene, d-limonene, γ-terpinene, α-terpinolene and p-cymene constituted more than 60% of total VOCs identified in carrots including “Atomic Red”, “Nantes”, “Cosmic Purple”, “Red Samurai”, “Eregli Black”, “White Satin”, “Parmex” and “Baby Carrot”. Thus, these cultivars may advise to carrot breeders due to the beneficial effects of terpenes, especially monoterpenes on health.

Deposition of carotenoids in egg yolk by short-term supplement of coloured carrot (Daucus carota) varieties as forage material for egg-laying hens.

Science.gov (United States)

Hammershøj, Marianne; Kidmose, Ulla; Steenfeldt, Sanna

2010-05-01

Supplying egg-laying hens with different forage materials may influence egg production and quality. The aim of this study was to examine the short-term effects of standard feed plus 70 g day(-1) per hen of three coloured carrot varieties (orange, yellow and purple) as forage material in comparison with a standard feed control on egg production, egg yolk colour and deposition of carotenoids in the yolk. Carrot supplementation reduced feed intakes significantly, but not on a dry matter basis. Orange carrot treatment significantly reduced egg mass production, whereas yellow and purple carrot treatments did not differ from the control. Egg and yolk weights of all carrot-supplemented treatments were significantly lower than those of the control, but yolk percentages were similar. Yolk redness increased significantly in the order control 1.5-fold) and beta-carotene (>100-fold) compared with the control. Supplementing the feed of egg-laying hens with coloured carrots efficiently increased yolk colour parameters and carotenoid contents, which gives opportunities for improved nutritional value of eggs from forage material-supplemented hens.

Perfusion based cell culture chips

DEFF Research Database (Denmark)

Heiskanen, Arto; Emnéus, Jenny; Dufva, Martin

2010-01-01

Performing cell culture in miniaturized perfusion chambers gives possibilities to experiment with cells under near in vivo like conditions. In contrast to traditional batch cultures, miniaturized perfusion systems provide precise control of medium composition, long term unattended cultures...... and tissue like structuring of the cultures. However, as this chapter illustrates, many issues remain to be identified regarding perfusion cell culture such as design, material choice and how to use these systems before they will be widespread amongst biomedical researchers....

Chromoplasts ultrastructure and estimated carotene content in root secondary phloem of different carrot varieties.

Science.gov (United States)

Kim, Ji Eun; Rensing, Kim H; Douglas, Carl J; Cheng, Kimberly M

2010-02-01

There have been few studies on quantifying carotenoid accumulation in carrots, and none have taken the comparative approach. The abundance and distribution of carotenes in carrot roots of three varieties, white, orange, and high carotene mass (HCM) were compared using light and transmission electron microscopy (TEM). Light microscopy has indicated that, in all three varieties, carotenes were most abundant in the secondary phloem and this area was selected for further TEM analysis. While carotenes were extracted during the fixation process for TEM, the high-pressure freezing technique we employed preserved the spaces (CS) left behind by the extracted carotene crystals. Chromoplasts from the HCM variety contained significantly (P chromoplasts from the orange variety. Chromoplasts from the white variety had few or no CS. There was no significant difference between the HCM and orange varieties in the number of chromoplasts per unit area, but the white variety had significantly (P chromoplasts than the other two varieties. A large number of starch-filled amyloplasts was observed in secondary phloem of the white variety but these were not found in the other two varieties. The results from this comparative approach clearly define the subcellular localization of carotenoids in carrot roots and suggest that while the HCM genotype was selectively bred for increased carotene content, this selection did not lead to increased numbers of carotene-containing chromoplasts but rather greater accumulation of carotene per chromoplast. Furthermore, the results confirm that roots of the white carrot variety retain residual amounts of carotene.

Development of Job’s tears ice cream recipes with carrot juice and pumpkin paste

Directory of Open Access Journals (Sweden)

Wiwat Wangcharoen

2011-11-01

Full Text Available Carrot juice and pumpkin paste were used as ingredients in Job’s tears ice cream. Carrot juice or pumpkin paste added at 50% was equally preferred by 100 consumers compared to the original Job’s tears ice cream. The new types of ice cream were lower in antioxidant capacity and higher in total phenolic content but could still be considered as potential antioxidant products. Purchase intent was significantly increased (p<0.05 if consumers were informed about the nutritional and antioxidant capacity of these products.

Effects of ionization and nitrous oxide on grated carrot respiration

International Nuclear Information System (INIS)

Chervin, C.

1992-06-01

Two treatments (nitrous oxide and irradiation) have been applied on grated carrots to reduce the respiratory crisis induced by wounding. Nitrous oxide inhibited cytochrome c oxidase; but, it neither diminished O 2 consumption of the tissues, nor modified atmospheres in a favourable way for conservation of grated carrots, stored in plastic bags (in the conditions chosen for this study). On the contrary, irradiation inhibited simultaneously the respiratory crisis and the ethylene production, both induced by wounding. This behaviour led to a lower consumption of sugars in irradiated tissues and to the generation of atmospheres, which were better adapted to the conservation needs (it was necessary to use plastic film with high permeability). Finally, an applied study demonstrated that irradiation, by permitting a less denaturing preparation than industrial process, allowed the conservation of produces with a better quality (nutritional, sensory and microbiological). Biochemical analyses have been validated by sensory analyses

Carnauba straw incorporated into the soil for fertilization carrot in organic cultivation

Directory of Open Access Journals (Sweden)

Micharlyson Carlos Morais

2017-10-01

Full Text Available Carrot is one of the most produced vegetables in Brazil and, with the growing demand for organic vegetables, it is necessary to develop production technologies that are less dependent on external inputs and more accessible to family agriculture. The objective of this study was to evaluate the effect of the addition to soil of carnauba straw as fertilizer, incorporated under different pre-planting periods, for the organic cultivation of the carrot. The experiment was carried out in the Experimental Horta of the Instituto Federal do Rio Grande do Norte, Campus Ipanguaçu, using the experimental design in randomized blocks with three replicates and five treatments referring to the times of 15; 30; 45 and 60 days for the incorporation of carnauba straw, in the amount of 14 t ha-1, prior to planting the carrot, and a control treatment without addition of straw to the soil. The evaluated characteristics was dry mass of the aerial part, the length and root diameter, and productivity. There was an effect of the incorporation time of the carnauba straw on the length and productivity, being the highest values observed when the straw was incorporated between 15 and 30 days before sowing.

Cell Culture Made Easy.

Science.gov (United States)

Dye, Frank J.

1985-01-01

Outlines steps to generate cell samples for observation and experimentation. The procedures (which use ordinary laboratory equipment) will establish a short-term primary culture of normal mammalian cells. Information on culture vessels and cell division and a list of questions to generate student interest and involvement in the topics are…

The 1064 nm laser-induced breakdown spectroscopy (LIBS) inspection to detect the nutrient elements in freshly cut carrot samples

Science.gov (United States)

Yudasari, N.; Prasetyo, S.; Suliyanti, M. M.

2018-03-01

The laser-induced breakdown spectroscopy (LIBS) technique was applied to detect the nutrient elements contained in fresh carrot. Nd:YAG laser the wavelength of 1064 nm was employed in the experiments for ablation. Employing simple set-up of LIBS and preparing the sample with less step method, we are able to detect 18 chemical elements including some fundamental element of carrot, i.e Mg, Al, Fe, Mn, Ti, Ca, and Mn. By applying normalized profiles calculation on some of the element, we are able to compare the concentration level of each element of the outer and inner part of carrot.

Irradiation treatment of minimally processed carrots for ensuring microbiological safety

Science.gov (United States)

Ashraf Chaudry, Muhammad; Bibi, Nizakat; Khan, Misal; Khan, Maazullah; Badshah, Amal; Jamil Qureshi, Muhammad

2004-09-01

Minimally processed fruits and vegetables are very common in developed countries and are gaining popularity in developing countries due to their convenience and freshness. However, minimally processing may result in undesirable changes in colour, taste and appearance due to the transfer of microbes from skin to the flesh. Irradiation is a well-known technology for elimination of microbial contamination. Food irradiation has been approved by 50 countries and is being applied commercially in USA. The purpose of this study was to evaluate the effect of irradiation on the quality of minimally processed carrots. Fresh carrots were peeled, sliced and PE packaged. The samples were irradiated (0, 0.5, 1.0, 2.0, 2.5, 3.0 kGy) and stored at 5°C for 2 weeks. The samples were analyzed for hardness, organoleptic acceptance and microbial load at 0, 7th and 15th day. The mean firmness of the control and all irradiated samples remained between 4.31 and 4.42 kg of force, showing no adverse effect of radiation dose. The effect of storage (2 weeks) was significant ( P10) in all other irradiated samples(1.0, 2.0, 2.5 and 3.0 kGy) after 2 weeks storage. No coliform or E. coli were detected in any of the samples (radiated or control) immediately after irradiation and during the entire storage period in minimally processed carrots. A dose of 2.0 kGy completely controlled the fungal and bacterial counts. The irradiated samples (2.0 kGy) were also acceptable sensorially.

Genomic identification of WRKY transcription factors in carrot (Daucus carota) and analysis of evolution and homologous groups for plants.

Science.gov (United States)

Li, Meng-Yao; Xu, Zhi-Sheng; Tian, Chang; Huang, Ying; Wang, Feng; Xiong, Ai-Sheng

2016-03-15

WRKY transcription factors belong to one of the largest transcription factor families. These factors possess functions in plant growth and development, signal transduction, and stress response. Here, we identified 95 DcWRKY genes in carrot based on the carrot genomic and transcriptomic data, and divided them into three groups. Phylogenetic analysis of WRKY proteins from carrot and Arabidopsis divided these proteins into seven subgroups. To elucidate the evolution and distribution of WRKY transcription factors in different species, we constructed a schematic of the phylogenetic tree and compared the WRKY family factors among 22 species, which including plants, slime mold and protozoan. An in-depth study was performed to clarify the homologous factor groups of nine divergent taxa in lower and higher plants. Based on the orthologous factors between carrot and Arabidopsis, 38 DcWRKY proteins were calculated to interact with other proteins in the carrot genome. Yeast two-hybrid assay showed that DcWRKY20 can interact with DcMAPK1 and DcMAPK4. The expression patterns of the selected DcWRKY genes based on transcriptome data and qRT-PCR suggested that those selected DcWRKY genes are involved in root development, biotic and abiotic stress response. This comprehensive analysis provides a basis for investigating the evolution and function of WRKY genes.

Influence of Cultivar and UGmax on Antioxidative Properties of Carrot Roots (Daucus Carota L. and their Stability During Freezing Process

Directory of Open Access Journals (Sweden)

Keutgen Anna J.

2014-12-01

Full Text Available In the present experiment, the significance of cultivar (convention-al and coloured and of the application of the soil fertility enhancer UGmax on health-promoting properties of carrot roots subjected to the freezing process of carrot cubes after water blanching was investigated. The selection of cultivar turned out to be highly signif-icant with respect to the development of health-promoting properties of carrot roots. The highest antioxidant properties were found in the purple cultivar ‘Deep Purple’. Its mean antioxidant capacity accounted for 5.31 mmol Fe+2 · kg–1 f.m. Essential for health-promoting properties were the contents of anthocyanins (R2 = 0.83, chlorogenic acid (R2 = 0.81 and total polyphenolics (R2 = 0.71. The application of the biological agent UGmax improved the qual-ity of carrot significantly, increasing the content of total carotenoids and reducing the losses of ascorbic acid during processing. The freezing process negatively influenced the antioxidative properties of carrot irrespective of cultivar and applied agro-technique (use of UGmax, especially in the case of water-soluble antioxidants such as anthocyanins and ascorbic acid.

A biocompatible micro cell culture chamber (mu CCC) for the culturing and on-line monitoring of eukaryote cells

DEFF Research Database (Denmark)

Stangegaard, Michael; Petronis, Sarunas; Jørgensen, Anders Michael

2006-01-01

culture chip compared to cell culture flasks. The cell culture chip could without further modification support cell growth of two other cell lines. Light coming from the microscope lamp during optical recordings of the cells was the only external factor identified, that could have a negative effect...... on cell survival. Low grade light exposure was however compatible with optical recordings as well as cell viability. These results strongly indicate that a cell culture chip could be constructed that allowed for on-line optical recording of cellular events without affecting the cell culturing condition...

A biocompatible micro cell culture chamber (microCCC) for the culturing and on-line monitoring of eukaryote cells

DEFF Research Database (Denmark)

Stangegaard, Michael; Petronis, Sarunas; Jørgensen, A M

2006-01-01

culture chip compared to cell culture flasks. The cell culture chip could without further modification support cell growth of two other cell lines. Light coming from the microscope lamp during optical recordings of the cells was the only external factor identified, that could have a negative effect...... on cell survival. Low grade light exposure was however compatible with optical recordings as well as cell viability. These results strongly indicate that a cell culture chip could be constructed that allowed for on-line optical recording of cellular events without affecting the cell culturing condition...

Identification of volatile organic compounds (VOCs) in different colour carrot (Daucus carota L.) cultivars using static headspace/gas chromatography/mass spectrometry

OpenAIRE

Zehra Güler; Fatih Karaca; Halit Yetisir

2015-01-01

Volatile organic compounds (VOCs) as well as sugar and acid contents affect carrot flavour. This study compared VOCs in 11 carrot cultivars. Gas chromatography/mass spectrometry using static headspace technique was applied to analyse the VOCs. The number of VOCs per sample ranged from 17 to 31. The primarily VOCs identified in raw carrots with the exception of “Yellow Stone” were terpenes, ranging from 65 to 95%. The monoterpenes with values ranging from 31 to 89% were higher than those (from...

Influence of culture media and environmental factors on mycelial growth and conidial production of Diplocarpon mali.

Science.gov (United States)

Zhao, H; Huang, L; Xiao, C L; Liu, J; Wei, J; Gao, X

2010-06-01

To identify media and environmental conditions suitable for rapid mycelial growth and sporulation of Diplocarpon mali. Liquid shake cultures were used to evaluate effects of media and environmental conditions on mycelial growth and conidial production of D. mali. Carrot sucrose broth (CSB), potato and carrot dextrose broth (PCDB) and potato and carrot sucrose broth (PCSB) were most favourable for rapid mycelial growth. PCDB, PCSB, PCB (potato and carrot broth) and carrot dextrose broth (CDB) were favourable for conidial production. All carbon sources tested and peptone favoured for mycelial growth. Carbon and nitrogen sources tested did not significantly stimulate conidial production. The optimum temperature for mycelial growth and conidial production was 25 degrees C. No mycelial growth occurred at 5 or 30 degrees C, but D. mali survived at these temperatures. Active mycelial growth occurred at pH 5-7, and pH 5-8 was favourable for sporulation. PCDB and PCSB incubated at 25 degrees C for 14 day are recommended for mycelial growth and conidial production of D. mali. The information generated in this study will facilitate mycological and pathological research on D. mali and Marssonina leaf blotch of apple caused by D. mali.

Combined effect of water loss and wounding stress on gene activation of metabolic pathways associated with phenolic biosynthesis in carrot

Directory of Open Access Journals (Sweden)

Alejandro eBecerra-Moreno

2015-10-01

Full Text Available Abstract: The application of postharvest abiotic stresses is an effective strategy to activate the primary and secondary metabolism of plants inducing the accumulation of antioxidant phenolic compounds. In the present study, the effect of water stress applied alone and in combination with wounding stress on the activation of primary (shikimic acid and secondary (phenylpropanoid metabolic pathways related with the accumulation of phenolic compound in plants was evaluated. Carrot (Daucus carota was used as model system for this study, and the effect of abiotic stresses was evaluated at the gene expression level and on the accumulation of metabolites. As control of the study, whole carrots were stored under the same conditions. Results demonstrated that water stress activated the primary and secondary metabolism of carrots, favoring the lignification process. Likewise, wounding stress induced higher activation of the primary and secondary metabolism of carrots as compared to water stress alone, leading to higher accumulation of shikimic acid, phenolic compounds and lignin. Additional water stress applied on wounded carrots exerted a synergistic effect on the wound-response at the gene expression level. For instance, when wounded carrots were treated with water stress, the tissue showed 20- and 14-fold increases in the relative expression of 3-deoxy-D-arabino-heptulosanate synthase and phenylalanine ammonia-lyase genes, respectively. However, since lignification was increased, lower accumulation of phenolic compounds was detected. Indicatively, at 48 h of storage, wounded carrots treated with water stress showed ~31% lower levels of phenolic compounds and ~23% higher lignin content as compared with wounded controls. In the present study, it was demonstrated that water stress is one of the pivotal mechanism of the wound-response in carrot. Results allowed the elucidation of strategies to induce the accumulation of specific primary or secondary

Studies in utilization of fertilizer and soil nitrogen by carrots

International Nuclear Information System (INIS)

Moussa, A.G.; Markgraf, G.; Geissler, T.

1985-01-01

Pot experiments were conducted to determine the extent of fertilizer N utilization by carrots, using double-labelled 15 N-ammonium nitrate. The degree of soil N utilization was also studied. The residual effect of nitrogen in the individual variants was determined in spinach grown as succeeding crop. Under the experimental conditions, N utilization was highest at high water supply (100 % of water capacity). Due to the daily rhythm of pot watering to approximately 100 % of water capacity, gas exchange (air and oxygen) was ensured as well, providing optimum growth conditions. At medium nitrogen rates (12.5 g N/m 2 ), carrots took up 44.5 % of the fertilizer N on sand and 54.5 % on loess soil. When water supply decreased to 70 % of the water capacity, utilization of fertilizer N declined to 26 % on sand and 43.8 % on loess soil. Spinach grown as succeeding crop took up more soil N than fertilizer N. (author)

Effect of gamma irradiation on microbial quality of minimally processed carrot and lettuce: A case study in Greater Accra region of Ghana

Science.gov (United States)

Frimpong, G. K.; Kottoh, I. D.; Ofosu, D. O.; Larbi, D.

2015-05-01

The effect of ionizing radiation on the microbiological quality on minimally processed carrot and lettuce was studied. The aim was to investigate the effect of irradiation as a sanitizing agent on the bacteriological quality of some raw eaten salad vegetables obtained from retailers in Accra, Ghana. Minimally processed carrot and lettuce were analysed for total viable count, total coliform count and pathogenic organisms. The samples collected were treated and analysed for a 15 day period. The total viable count for carrot ranged from 1.49 to 14.01 log10 cfu/10 g while that of lettuce was 0.70 to 8.5 7 log10 cfu/10 g. It was also observed that total coliform count for carrot was 1.46-7.53 log10 cfu/10 g and 0.14-7.35 log10 cfu/10 g for lettuce. The predominant pathogenic organisms identified were Bacillus cereus, Cronobacter sakazakii, Staphylococcus aureus, and Klebsiella spp. It was concluded that 2 kGy was most effective for medium dose treatment of minimally processed carrot and lettuce.

Irradiation treatment of minimally processed carrots for ensuring microbiological safety

International Nuclear Information System (INIS)

Ashraf Chaudry, Muhammad; Bibi, Nizakat; Khan, Misal; Khan, Maazullah; Badshah, Amal; Jamil Qureshi, Muhammad

2004-01-01

Minimally processed fruits and vegetables are very common in developed countries and are gaining popularity in developing countries due to their convenience and freshness. However, minimally processing may result in undesirable changes in colour, taste and appearance due to the transfer of microbes from skin to the flesh. Irradiation is a well-known technology for elimination of microbial contamination. Food irradiation has been approved by 50 countries and is being applied commercially in USA. The purpose of this study was to evaluate the effect of irradiation on the quality of minimally processed carrots. Fresh carrots were peeled, sliced and PE packaged. The samples were irradiated (0, 0.5, 1.0, 2.0, 2.5, 3.0 kGy) and stored at 5 deg. C for 2 weeks. The samples were analyzed for hardness, organoleptic acceptance and microbial load at 0, 7th and 15th day. The mean firmness of the control and all irradiated samples remained between 4.31 and 4.42 kg of force, showing no adverse effect of radiation dose. The effect of storage (2 weeks) was significant (P 5 cfu/g, 3.0x10 2 and few colonies(>10) in all other irradiated samples(1.0, 2.0, 2.5 and 3.0 kGy) after 2 weeks storage. No coliform or E. coli were detected in any of the samples (radiated or control) immediately after irradiation and during the entire storage period in minimally processed carrots. A dose of 2.0 kGy completely controlled the fungal and bacterial counts. The irradiated samples (2.0 kGy) were also acceptable sensorially

Cell Culturing of Cytoskeleton

Science.gov (United States)

2004-01-01

Biomedical research offers hope for a variety of medical problems, from diabetes to the replacement of damaged bone and tissues. Bioreactors, which are used to grow cells and tissue cultures, play a major role in such research and production efforts. Cell culturing, such as this bone cell culture, is an important part of biomedical research. The BioDyn payload includes a tissue engineering investigation. The commercial affiliate, Millenium Biologix, Inc., has been conducting bone implant experiments to better understand how synthetic bone can be used to treat bone-related illnesses and bone damaged in accidents. On STS-95, the BioDyn payload will include a bone cell culture aimed to help develop this commercial synthetic bone product. Millenium Biologix, Inc., is exploring the potential for making human bone implantable materials by seeding its proprietary artificial scaffold material with human bone cells. The product of this tissue engineering experiment using the Bioprocessing Modules (BPMs) on STS-95 is space-grown bone implants, which could have potential for dental implants, long bone grafts, and coating for orthopedic implants such as hip replacements.

9 CFR 101.6 - Cell cultures.

Science.gov (United States)

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Cell cultures. 101.6 Section 101.6..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS DEFINITIONS § 101.6 Cell cultures. When used in conjunction with or in reference to cell cultures, which may be referred to as tissue cultures...

Effect of gamma irradiation on microbial quality of minimally processed carrot and lettuce: A case study in Greater Accra region of Ghana

International Nuclear Information System (INIS)

Frimpong, G.K.; Kottoh, I.D.; Ofosu, D.O.; Larbi, D.

2015-01-01

The effect of ionizing radiation on the microbiological quality on minimally processed carrot and lettuce was studied. The aim was to investigate the effect of irradiation as a sanitizing agent on the bacteriological quality of some raw eaten salad vegetables obtained from retailers in Accra, Ghana. Minimally processed carrot and lettuce were analysed for total viable count, total coliform count and pathogenic organisms. The samples collected were treated and analysed for a 15 day period. The total viable count for carrot ranged from 1.49 to 14.01 log 10 cfu/10 g while that of lettuce was 0.70 to 8.5 7 log 10 cfu/10 g. It was also observed that total coliform count for carrot was 1.46–7.53 log 10 cfu/10 g and 0.14–7.35 log 10 cfu/10 g for lettuce. The predominant pathogenic organisms identified were Bacillus cereus, Cronobacter sakazakii, Staphylococcus aureus, and Klebsiella spp. It was concluded that 2 kGy was most effective for medium dose treatment of minimally processed carrot and lettuce. - Highlights: • The microbial load on the cut-vegetables was beyond acceptable level for consumption. • The microbial contamination of carrot was found to be higher than that of lettuce. • 2 kGy was most appropriate in treating cut-vegetables for microbial safety

Statistical Quality Assessment of Pre-fried Carrots Using Multispectral Imaging

DEFF Research Database (Denmark)

Sharifzadeh, Sara; Clemmensen, Line Katrine Harder; Løje, Hanne

2013-01-01

Multispectral imaging is increasingly being used for quality assessment of food items due to its non-invasive benefits. In this paper, we investigate the use of multispectral images of pre-fried carrots, to detect changes over a period of 14 days. The idea is to distinguish changes in quality from...

The fungi causin damping-off of carrot seedlings

Directory of Open Access Journals (Sweden)

Bogdan Nowicki

2013-12-01

Full Text Available When 136 samples of dying carrot seedlings from several fields were analyzed Alternaria rudicina proved to be the most common seedling pathogen (41%, followed by some Fusarium species (27%, mostly F. avenaceum.The less common seedling pathogens were Pythium spp. (13%, Phoma spp.(2,5% and Botrytis cinerea (1,4%. Some other fungi (Bipolaris sorokiniana, Sclerotinia sclerotiorum, Stemphylium botryosym and Ulocladium consortiale were found in less than 1% of seedlings examined.

Raman spectroscopy application in frozen carrot cooked in different ways and the relationship with carotenoids.

Science.gov (United States)

Camorani, Paolo; Chiavaro, Emma; Cristofolini, Luigi; Paciulli, Maria; Zaupa, Maria; Visconti, Attilio; Fogliano, Vincenzo; Pellegrini, Nicoletta

2015-08-30

Raman spectroscopy, in its confocal micro-Raman variation, has been recently proposed as a spatially resolved method to identify carotenoids in various food matrices, being faster, non-destructive, and avoiding sample extraction, but no data are present in the literature concerning its application to the evaluation of carotenoid pattern changes after thermal treatment of carrots. The effect of three cooking methods (i.e. boiling, steaming and microwaving) was evaluated on frozen carrot, comparing changes on carotenoid profiles measured by means of Raman spectroscopy with their high-performance liquid chromatographic determination and colour. A more pronounced detrimental effect on carotenoids was detected in steamed carrots, in accordance with colour data. Conversely, boiling and, to a lesser extent, microwaving caused an increase in carotenoid concentration. Cooking procedures affected the Raman spectral features of carotenoids, causing a shift of vibration frequencies towards a higher energy, increase in the spectral baseline and peak intensities as well as a broadening of their width, probably in relation to the thermal degradation of longer carotenoids (i.e. the all-trans form) and the isomerization process. In particular, steamed samples showed a significantly higher increase of centre frequency, in accordance with a more pronounced isomerization and changes in colour parameters. This work showed that the evolution of Raman spectral parameters could provide information on carotenoid bioaccessibility for carrots cooked using various methods. This paves the way for a future use of this technique to monitor and optimize cooking processes aimed at maximizing carotenoid bioaccessibility and bioavailability. © 2014 Society of Chemical Industry.

Irradiation treatment of minimally processed carrots for ensuring microbiological safety

Energy Technology Data Exchange (ETDEWEB)

Ashraf Chaudry, Muhammad; Bibi, Nizakat; Khan, Misal; Khan, Maazullah; Badshah, Amal; Jamil Qureshi, Muhammad

2004-10-01

Minimally processed fruits and vegetables are very common in developed countries and are gaining popularity in developing countries due to their convenience and freshness. However, minimally processing may result in undesirable changes in colour, taste and appearance due to the transfer of microbes from skin to the flesh. Irradiation is a well-known technology for elimination of microbial contamination. Food irradiation has been approved by 50 countries and is being applied commercially in USA. The purpose of this study was to evaluate the effect of irradiation on the quality of minimally processed carrots. Fresh carrots were peeled, sliced and PE packaged. The samples were irradiated (0, 0.5, 1.0, 2.0, 2.5, 3.0 kGy) and stored at 5 deg. C for 2 weeks. The samples were analyzed for hardness, organoleptic acceptance and microbial load at 0, 7th and 15th day. The mean firmness of the control and all irradiated samples remained between 4.31 and 4.42 kg of force, showing no adverse effect of radiation dose. The effect of storage (2 weeks) was significant (P< 0.05) with values ranging between 4.28 and 4.39 kg of force. The total bacterial counts at 5 deg. C for non-irradiated and 0.5 kGy irradiated samples were 6.3x10{sup 5} cfu/g, 3.0x10{sup 2} and few colonies(>10) in all other irradiated samples(1.0, 2.0, 2.5 and 3.0 kGy) after 2 weeks storage. No coliform or E. coli were detected in any of the samples (radiated or control) immediately after irradiation and during the entire storage period in minimally processed carrots. A dose of 2.0 kGy completely controlled the fungal and bacterial counts. The irradiated samples (2.0 kGy) were also acceptable sensorially.

Comparison of carrot (Daucus carota drying in microwave and in vacuum microwave

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R. Béttega

2014-06-01

Full Text Available Drying is a single operation employed to prolong the life of a large quantity of vegetables. Carrot (Daucus carota drying has been the subject of many studies. This plant has been highlighted in the human diet for having high nutritional value, mainly due to the high content of β-carotene. In this work, carrot drying behavior was studied in a regular microwave dryer and a vacuum microwave dryer. A vacuum of 450 mmHg was applied for drying of carrot in different geometrical shapes (cubes, discs and sticks. The samples were dried at power ratings of 1.0 W/g, 1.5 W/g and 2.0 W/g for both methods of drying. The evolution of physical properties such as density, volume and porosity was monitored and related to the moisture content of the sample and to the method of drying and power rating used. The geometric shape of the sample influenced the drying kinetics and it was verified that the cubic form was responsible for a slower drying. The application of vacuum showed no major changes in the drying kinetics in microwave but influenced the physical properties of the material. The influence of power ratings on the content of β-carotene was also evaluated and discussed. The main difference observed was the lower shrinkage of the samples dried in the vacuum microwave compared to those dried only in microwave.

Microfluidic cell culture systems for drug research.

Science.gov (United States)

Wu, Min-Hsien; Huang, Song-Bin; Lee, Gwo-Bin

2010-04-21

In pharmaceutical research, an adequate cell-based assay scheme to efficiently screen and to validate potential drug candidates in the initial stage of drug discovery is crucial. In order to better predict the clinical response to drug compounds, a cell culture model that is faithful to in vivo behavior is required. With the recent advances in microfluidic technology, the utilization of a microfluidic-based cell culture has several advantages, making it a promising alternative to the conventional cell culture methods. This review starts with a comprehensive discussion on the general process for drug discovery and development, the role of cell culture in drug research, and the characteristics of the cell culture formats commonly used in current microfluidic-based, cell-culture practices. Due to the significant differences in several physical phenomena between microscale and macroscale devices, microfluidic technology provides unique functionality, which is not previously possible by using traditional techniques. In a subsequent section, the niches for using microfluidic-based cell culture systems for drug research are discussed. Moreover, some critical issues such as cell immobilization, medium pumping or gradient generation in microfluidic-based, cell-culture systems are also reviewed. Finally, some practical applications of microfluidic-based, cell-culture systems in drug research particularly those pertaining to drug toxicity testing and those with a high-throughput capability are highlighted.

Esplanada: cultivar de cenoura de verão para fins de processamento Esplanada: a new tropical carrot variety suitable for minimum processing

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Jairo V. Vieira

2005-07-01

Full Text Available A cenoura é uma das hortaliças tradicionais mais consumidas no Brasil. Entretanto, o consumo de mini cenouras tipo "cenourete" obtidas via processamento mínimo ainda é muito pequeno no país. Isto se deve possivelmente ao restrito conhecimento da tecnologia de produção deste produto e da dificuldade em se obter matéria prima de qualidade e com regularidade, especialmente no período de verão. As raízes das principais cultivares utilizadas no Brasil não apresentam características de qualidade como coloração, formato e tamanho adequados para possibilitar máximo rendimento industrial. Neste contexto, foi desenvolvida a cultivar de polinização aberta "Esplanada", que apresenta boa adaptação às condições edafoclimáticas brasileiras, possui alta resistência à queima-das-folhas, baixa incidência de florescimento precoce no verão e resistência moderada a nematóides formadores de galhas. Estas características viabilizam seu cultivo em qualquer época do ano, nas principais regiões de produção. As raízes são compridas, finas e apresentam coloração uniforme, características estas adequadas para o processamento mínimo visando a produção de cenourete. Esta cultivar poderá ocupar posição estratégica na cadeia produtiva de cenoura, uma vez que vai possibilitar a produção de mini cenouras durante todo o ano em qualquer região do país.Carrot is one of the most consumed vegetables in Brazil. However, the consumption of "baby carrots", obtained by minimally processed roots, is still very low in the country, due to lack of processing technology knowledge and the low amount of minimally processed carrots produced in Brazil as a consequence of the low industrial productivity. The carrot varieties currently employed in minimum processing have as major disadvantages a series of low quality attributes including inadequate root shape and size which precludes reaching the maximum industrial yield. In this context, Embrapa

Use of an adaptable cell culture kit for performing lymphocyte and monocyte cell cultures in microgravity

Science.gov (United States)

Hatton, J. P.; Lewis, M. L.; Roquefeuil, S. B.; Chaput, D.; Cazenave, J. P.; Schmitt, D. A.

1998-01-01

The results of experiments performed in recent years on board facilities such as the Space Shuttle/Spacelab have demonstrated that many cell systems, ranging from simple bacteria to mammalian cells, are sensitive to the microgravity environment, suggesting gravity affects fundamental cellular processes. However, performing well-controlled experiments aboard spacecraft offers unique challenges to the cell biologist. Although systems such as the European 'Biorack' provide generic experiment facilities including an incubator, on-board 1-g reference centrifuge, and contained area for manipulations, the experimenter must still establish a system for performing cell culture experiments that is compatible with the constraints of spaceflight. Two different cell culture kits developed by the French Space Agency, CNES, were recently used to perform a series of experiments during four flights of the 'Biorack' facility aboard the Space Shuttle. The first unit, Generic Cell Activation Kit 1 (GCAK-1), contains six separate culture units per cassette, each consisting of a culture chamber, activator chamber, filtration system (permitting separation of cells from supernatant in-flight), injection port, and supernatant collection chamber. The second unit (GCAK-2) also contains six separate culture units, including a culture, activator, and fixation chambers. Both hardware units permit relatively complex cell culture manipulations without extensive use of spacecraft resources (crew time, volume, mass, power), or the need for excessive safety measures. Possible operations include stimulation of cultures with activators, separation of cells from supernatant, fixation/lysis, manipulation of radiolabelled reagents, and medium exchange. Investigations performed aboard the Space Shuttle in six different experiments used Jurkat, purified T-cells or U937 cells, the results of which are reported separately. We report here the behaviour of Jurkat and U937 cells in the GCAK hardware in ground

Elimination of Listeria inoculated in ready-to-eat carrots by combination of antimicrobial coating and γ-irradiation

International Nuclear Information System (INIS)

Turgis, Mélanie; Millette, Mathieu; Salmieri, Stéphane; Lacroix, Monique

2012-01-01

A combined treatment of an edible coating composed of trans-cinnamaldehyde (TCN; 0.5% p/p) with γ-irradiation was investigated against Listeria inoculated in peeled mini-carrots. First, the D 10 value (γ-irradiation dose required to eliminate 90% of the bacterial population) of TCN was evaluated under air. This treatment resulted in a 3.66-fold increase in relative bacterial radiosensitivity (RBR) as compared to the control without antimicrobial coating. Secondly, the shelf life of mini-carrots during 21 day of storage at 4 °C was studied. Antimicrobial coating containing TCN was assayed in combination with two irradiation doses (0.25 and 0.5 kGy). Results suggested that the inactive coating did not have any antimicrobial effect against Listeria while the coating containing TCN resulted in a 1.29 log reduction in carrots packed under air after 21 days of storage. Hence, these observations indicated that the combination of irradiation with antimicrobial coating played an important role in enhancing the radiosensitization of Listeria to γ-irradiation. - Highlights: ► Synergistic effect of essential oils and γ-radiation against food pathogens. ► Reducing any undesirable organoleptic impact due to high concentration of EOs. ► Potential in controlling food pathogens and food spoilage bacteria in food. ► Elimination of Listeria monocytogenes in the carrots during the storage.

Influence of Cultivars and Seed Thermal Treatment on the Development of Fungal Pathogens in Carrot and Onion Plants

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Martin Koudela

2016-01-01

Full Text Available Carrot and onion are vegetables representing an important segment of fresh market. They suffer from serious fungal diseases that can inflict great damage on crops, i.e. alternaria leaf blight, peronospora downy mildew, and botrytis neck rot. The resistance of selected carrot and onion cultivars important for the production of vegetables in the Czech Republic was tested by exposure to targeted infection by the above fungal pathogens. The exposure of eleven carrot cultivars to spores of Alternaria dauci showed that the most resistant and sensitive cultivars were Katrin, Cortina F1, Afalon F1 and Favorit, Tinga, Berlika F1, respectively. A targeted infection of onion cultivars with Botrytis aclada clustered them into three groups: Amfora F1, Bolero, Tosca, Triumf F1 (strong resistance, Avalon, Grenada (medium resistance, Alice, Karmen, Všetana (low resistance. Similar groups were distinguished also after the infection with Peronospora destructor: Avalon, Bolero, Tosca (strong resistance, Alice, Amfora F1, Grenada, Karmen, Triumf F1 (medium resistance,Všetana (low resistance. Hot water treatment of carrot seeds applied after the inoculation with A. dauci decreased the development of the infection 1.3-2.3-fold, whereas the protective effect observed with onion seeds against the infection by P. destructor and B. aclada was lower.

The effect of osmotic pretreatment on the density of hot-air-dried carrot

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J Soleimani

2012-02-01

Full Text Available Consumption of large amounts of fruits and vegetables throughout the world, have encouraged the development of various methods for their processing. Drying is considers as the most common method for preservation of vegetable and fruits. Although drying extend the shelf-life, it has various side effects on keeping quality of these foods; including decreasing of the color and texture quality as well as missing the flavor and nutritional values. These negative effects have increased the demand for the discovering the alternative drying methods and consequently for the production of fresh-like products. The aim of this study was to introduce and optimize the novel method for the drying of carrot as well as to develop and optimize the quality of osmo-air-dried carrots with special respect to the color, flavor, texture, rehydration properties, density and shriveling of the product. For this, the effect of osmotic pretreatment on the density of carrot slices was investigated, using 50% glucose syrup +5% salt at 40°C with 150 rpm, followed by complementary drying step. The result of treated group was compared with control samples which were dried only by hot-air-drier. The results showed that using osmotic pretreatment could increase the density through inhibition of the product's shrinkage. Meanwhile, in air-dried samples the density was decreased considerably and high shrinkage was also observed.

Principles of cancer cell culture.

Science.gov (United States)

Cree, Ian A

2011-01-01

The basics of cell culture are now relatively common, though it was not always so. The pioneers of cell culture would envy our simple access to manufactured plastics, media and equipment for such studies. The prerequisites for cell culture are a well lit and suitably ventilated laboratory with a laminar flow hood (Class II), CO(2) incubator, benchtop centrifuge, microscope, plasticware (flasks and plates) and a supply of media with or without serum supplements. Not only can all of this be ordered easily over the internet, but large numbers of well-characterised cell lines are available from libraries maintained to a very high standard allowing the researcher to commence experiments rapidly and economically. Attention to safety and disposal is important, and maintenance of equipment remains essential. This chapter should enable researchers with little prior knowledge to set up a suitable laboratory to do basic cell culture, but there is still no substitute for experience within an existing well-run laboratory.

The use of tissue culture techniques to detect irradiated vegetables

International Nuclear Information System (INIS)

Al-Safadi, B.; Sharabi, N.E.; Nabulsi, I

2001-01-01

the ability of two tissue culture methods, callus and vegetable growth induction, to detect irradiated vegetables was evaluated. Potato tubers, carrot roots, garlic cloves and onion bulbs were subjected to various gamma radiation doses (0, 25, 100, 150, 250, 500, 750, and 1000 Gy). Irradiated vegetables were cultured in vitro and in vivo (pots). Gamma irradiation significantly reduced callus-forming ability especially in carrot and potato where no callus was observed in doses higher than 50 Gy. Length of shoots and roots growing from irradiated garlic and onion explants was considerably reduced starting from the 25 Gy dose. No roots were formed on garlic explants at any irradiation dose. Garlic leaves growing from irradiated explants were spotted with purple to brown spots. The intensity of these spots increased as gamma ray dosage increased. In the pot experiment, potato plant appeared in the control only. On the contrary, a complete sprouting of garlic and onion was seen in all irradiation treatments. It was not possible to distinguish between the various irradiation treatments and the control 3 days after planting in pots. The two in vitro techniques, tested in our study, may effectively be used to detect irradiated vegetables and estimate the range of doses used. The callus formation method is more useful for potato and carrot, since regeneration of shoots in vitro from these two plants takes along time, making this method unpractical. The other technique is very useful in the case of onion and garlic since it is rapid. The two techniques can be used with most of the vegetables that can be cultured in vitro. (Author)

Mobilizing cities towards a low-carbon future: Tambourines, carrots and sticks

International Nuclear Information System (INIS)

Azevedo, Isabel; Delarue, Erik; Meeus, Leonardo

2013-01-01

In the transition towards a low-carbon future in Europe, cities' actions are of major importance due to the prominence of urbanization, both in terms of population and in terms of greenhouse gas (GHG) emissions. As a result, we need city authorities to act, by using their competences as policy makers as well as energy users. However, cities are still not moving as fast as one might expect, indicating the need for additional incentives to prompt local action. Therefore, the aim of this paper is to present an overview of external incentives that might prompt cities to act and to highlight good practices that could be used in future initiatives. This paper first discusses how to evaluate the climate and energy performance of a city and how local authorities can contribute to its improvements. Moreover, it analyses the disincentives that local governments are confronted with, categorizing them as simple market failures, institutional failures and multi-agent failures. The paper then presents a survey of initiatives at national and EU levels to promote local action towards a low-carbon future; grouping them into tambourines, carrots and sticks. We focus on Austria, Germany, the Netherlands and Sweden because they are pioneering countries regarding energy policies for cities. - Highlights: • CO 2 and energy consumption could be used to evaluate the performance of cities. • Simple market, institutional and multi-agent failures are hampering local action. • National and EU actions use mainly tambourines and carrots, rather than sticks. • Covenant of Mayors is one of the most noteworthy tambourine type of instruments. • Carrots notable features are: use of competition and involvement of third-parties

High Pressure Processing Treatment of Fresh-Cut Carrots: Effect of Presoaking in Calcium Salts on Quality Parameters

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Yong Yu

2018-01-01

Full Text Available Effect of high pressure (HP treatment (200–600 MPa; 0–20 min on quality of fresh-cut carrot slices was evaluated after presoaking in selected calcium salt solutions (1% calcium chloride, 1% calcium lactate, 1% calcium gluconate, and distilled water as control for one hour. Results showed that calcium chloride (CaCl2 solution pretreatment was most effective for preserving the hardness of carrot slices at 400 and 600 MPa and this treatment also resulted in the least amount of color change in carrots, followed by calcium lactate, gluconate, and control pretreatments. The average sensory evaluation scores during 9 days of refrigerated storage at 4°C in control, CaCl2, calcium lactate, and calcium gluconate presoaking treatments followed by HP treatment were 6.4 ± 0.5, 8.0 ± 0.5, 7.8 ± 0.4, and 7.6 ± 0.3, respectively, on a zero to 9 scale for quality.

Bacterial cell culture

OpenAIRE

sprotocols

2014-01-01

### Materials 1. Glass culture tubes with metal caps and labels - Growth medium, from media room or customized - Glass pipette tubes - Parafilm ### Equipment 1. Vortexer - Fireboy or Bunsen burner - Motorized pipette - Micropipettes and sterile tips ### Procedure For a typical liquid culture, use 5 ml of appropriate medium. The amount in each tube does not have to be exact if you are just trying to culture cells for their precious DNA. 1. Streak an a...

Replication of cultured lung epithelial cells

International Nuclear Information System (INIS)

Guzowski, D.; Bienkowski, R.

1986-01-01

The authors have investigated the conditions necessary to support replication of lung type 2 epithelial cells in culture. Cells were isolated from mature fetal rabbit lungs (29d gestation) and cultured on feeder layers of mitotically inactivated 3T3 fibroblasts. The epithelial nature of the cells was demonstrated by indirect immunofluorescent staining for keratin and by polyacid dichrome stain. Ultrastructural examination during the first week showed that the cells contained myofilaments, microvilli and lamellar bodies (markers for type 2 cells). The following changes were observed after the first week: increase in cell size; loss of lamellar bodies and appearance of multivesicular bodies; increase in rough endoplasmic reticulum and golgi; increase in tonafilaments and well-defined junctions. General cell morphology was good for up to 10 wk. Cells cultured on plastic surface degenerated after 1 wk. Cell replication was assayed by autoradiography of cultures exposed to ( 3 H)-thymidine and by direct cell counts. The cells did not replicate during the first week; however, between 2-10 wk the cells incorporated the label and went through approximately 6 population doublings. They have demonstrated that lung alveolar epithelial cells can replicate in culture if they are maintained on an appropriate substrate. The coincidence of ability to replicate and loss of markers for differentiation may reflect the dichotomy between growth and differentiation commonly observed in developing systems

Essential-oil composition of Daucus carota ssp. major (Pastinocello Carrot) and nine different commercial varieties of Daucus carota ssp. sativus fruits.

Science.gov (United States)

Flamini, Guido; Cosimi, Elena; Cioni, Pier Luigi; Molfetta, Ilaria; Braca, Alessandra

2014-07-01

The chemical composition of the essential oils obtained by hydrodistillation from the pastinocello carrot, Daucus carota ssp. major (Vis.) Arcang. (flowers and achenes), and from nine different commercial varieties of D. carota L. ssp. sativus (achenes) was investigated by GC/MS analyses. Selective breeding over centuries of a naturally occurring subspecies of the wild carrot, D. carota L. ssp. sativus, has produced the common garden vegetable with reduced bitterness, increased sweetness, and minimized woody core. On the other hand, the cultivation of the pastinocello carrot has been abandoned, even if, recently, there has been renewed interest in the development of this species, which risks genetic erosion. The cultivated carrot (D. carota ssp. sativus) and the pastinocello carrot (D. carota ssp. major) were classified as different subspecies of the same species. This close relationship between the two subspecies urged us to compare the chemical composition of their essential oils, to evaluate the differences. The main essential-oil constituents isolated from the pastinocello fruits were geranyl acetate (34.2%), α-pinene (12.9%), geraniol (6.9%), myrcene (4.7%), epi-α-bisabolol (4.5%), sabinene (3.3%), and limonene (3.0%). The fruit essential oils of the nine commercial varieties of D. carota ssp. sativus were very different from that of pastinocello, as also confirmed by multivariate statistical analyses. Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.

Exploring the Effects of Pulsed Electric Field Processing Parameters on Polyacetylene Extraction from Carrot Slices

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Ingrid Aguiló-Aguayo

2015-03-01

Full Text Available The effects of various pulsed electric field (PEF parameters on the extraction of polyacetylenes from carrot slices were investigated. Optimised conditions with regard to electric field strength (1–4 kV/cm, number of pulses (100–1500, pulse frequency (10–200 Hz and pulse width (10–30 μs were identified using response surface methodology (RSM to maximise the extraction of falcarinol (FaOH, falcarindiol (FaDOH and falcarindiol-3-acetate (FaDOAc from carrot slices. Data obtained from RSM and experiments fitted significantly (p < 0.0001 the proposed second-order response functions with high regression coefficients (R2 ranging from 0.82 to 0.75. Maximal FaOH (188%, FaDOH (164.9% and FaDOAc (166.8% levels relative to untreated samples were obtained from carrot slices after applying PEF treatments at 4 kV/cm with 100 number of pulses of 10 μs at 10 Hz. The predicted values from the developed quadratic polynomial equation were in close agreement with the actual experimental values with low average mean deviations (E% ranging from 0.68% to 3.58%.

Reversible gelling culture media for in-vitro cell culture in three-dimensional matrices

Science.gov (United States)

An, Yuehuei H.; Mironov, Vladimir A.; Gutowska, Anna

2000-01-01

A gelling cell culture medium useful for forming a three dimensional matrix for cell culture in vitro is prepared by copolymerizing an acrylamide derivative with a hydrophilic comonomer to form a reversible (preferably thermally reversible) gelling linear random copolymer in the form of a plurality of linear chains having a plurality of molecular weights greater than or equal to a minimum gelling molecular weight cutoff, mixing the copolymer with an aqueous solvent to form a reversible gelling solution and adding a cell culture medium to the gelling solution to form the gelling cell culture medium. Cells such as chondrocytes or hepatocytes are added to the culture medium to form a seeded culture medium, and temperature of the medium is raised to gel the seeded culture medium and form a three dimensional matrix containing the cells. After propagating the cells in the matrix, the cells may be recovered by lowering the temperature to dissolve the matrix and centrifuging.

Traditional and Modern Cell Culture in Virus Diagnosis.

Science.gov (United States)

Hematian, Ali; Sadeghifard, Nourkhoda; Mohebi, Reza; Taherikalani, Morovat; Nasrolahi, Abbas; Amraei, Mansour; Ghafourian, Sobhan

2016-04-01

Cell cultures are developed from tissue samples and then disaggregated by mechanical, chemical, and enzymatic methods to extract cells suitable for isolation of viruses. With the recent advances in technology, cell culture is considered a gold standard for virus isolation. This paper reviews the evolution of cell culture methods and demonstrates why cell culture is a preferred method for identification of viruses. In addition, the advantages and disadvantages of both traditional and modern cell culture methods for diagnosis of each type of virus are discussed. Detection of viruses by the novel cell culture methods is considered more accurate and sensitive. However, there is a need to include some more accurate methods such as molecular methods in cell culture for precise identification of viruses.

Thermal time model for Egyptian broomrape ‎‎(Phelipanche aegyptiaca parasitism dynamics in carrot ‎‎(Daucus carota L.: Field validation ‎

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Amnon Cochavi

2016-12-01

Full Text Available Carrot, a highly profitable crop in Israel, is severely damaged by Phelipanche aegyptiaca ‎parasitism. Herbicides can effectively control the parasite and prevent damage, but for ‎optimal results, knowledge about the soil-subsurface phenological stage of the parasite is ‎essential. Parasitism dynamics models have been successfully developed for the parasites P. ‎aegyptiaca, Orobanche cumana and O. minor in the summer crops, tomato, sunflower and ‎red clover, respectively. However, these models, which are based on a linear relationship ‎between thermal time and the parasitism dynamics, may not necessarily be directly ‎applicable to the P. aegyptiaca-carrot system. The objective of the current study was to ‎develop a thermal time model to predict the effect of P. aegyptiaca parasitism dynamics on ‎carrot growth. For development and validation of the models, data was collected from a ‎temperature-controlled growth experiment and from 13 plots naturally infested with P. ‎aegyptiaca in commercial carrot fields. Our results revealed that P. aegyptiaca development ‎is related to soil temperature. Moreover, unlike P. aegyptiaca parasitism in sunflower and ‎tomato, which could be predicted both a linear model, P. aegyptiaca parasitism dynamics ‎on carrot roots required a nonlinear model, due to the wider range of growth temperatures of ‎both the carrot and the parasite. Hence, two different nonlinear models were developed for ‎optimizing the prediction of P. aegyptiaca parasitism dynamics. Both models, a beta ‎function model and combined model composed of a beta function and a sigmoid curve, were ‎able to predict first P. aegyptiaca attachment. However, overall P. aegyptiaca dynamics was ‎described more accurately by the combined model (RMSE =14.58 and 10.79, respectively. ‎The results of this study will complement previous studies on P. aegyptiaca management by ‎herbicides to facilitate optimal carrot growth and

Application of cell co-culture system to study fat and muscle cells.

Science.gov (United States)

Pandurangan, Muthuraman; Hwang, Inho

2014-09-01

Animal cell culture is a highly complex process, in which cells are grown under specific conditions. The growth and development of these cells is a highly unnatural process in vitro condition. Cells are removed from animal tissues and artificially cultured in various culture vessels. Vitamins, minerals, and serum growth factors are supplied to maintain cell viability. Obtaining result homogeneity of in vitro and in vivo experiments is rare, because their structure and function are different. Living tissues have highly ordered complex architecture and are three-dimensional (3D) in structure. The interaction between adjacent cell types is quite distinct from the in vitro cell culture, which is usually two-dimensional (2D). Co-culture systems are studied to analyze the interactions between the two different cell types. The muscle and fat co-culture system is useful in addressing several questions related to muscle modeling, muscle degeneration, apoptosis, and muscle regeneration. Co-culture of C2C12 and 3T3-L1 cells could be a useful diagnostic tool to understand the muscle and fat formation in animals. Even though, co-culture systems have certain limitations, they provide a more realistic 3D view and information than the individual cell culture system. It is suggested that co-culture systems are useful in evaluating the intercellular communication and composition of two different cell types.

Path analysis suggests phytoene accumulation is the key step limiting the carotenoid pathway in white carrot roots

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Carlos Antonio Fernandes Santos

2005-01-01

Full Text Available Two F2 carrot (Daucus carota L. populations (orange rooted Brasilia x very dark orange rooted High Carotene Mass - HCM cross and the dark orange rooted cultivated variety B493 x white rooted wild carrot Queen Anne's Lace - QAL cross with very unrelated genetic backgrounds were used to investigate intrinsic factors limiting carotenoid accumulation in carrots by applying phenotypic correlation and path analysis to study the relationships between major root carotenes, root color and several other morphological traits. Most of the correlations between traits were close and agreed in sign between the two populations. Root weight had a moderate to highly significant positive correlation with leaf length, root length and top and middle root diameter. Although phenotypic correlations failed to identify the order of the substrates and products in the carotenoid pathway the correct order of substrates and products (phytoene -> zeta-carotene -> lycopene was identified in the causal diagram of beta-carotene for the Brasilia x HCM population. Path analysis of beta-carotene synthesis in the B493 x QAL population suggested that selection for root carotenes had little effect on plant morphological traits. Causal model of beta-carotene and lycopene in the B493 x QAL population suggested that phytoene synthesis is the key step limiting the carotenoid pathway in white carrots. Path analysis, first presented by Sewall Wright to study quantitative traits, appears to be a powerful statistical approach for the identification of key compounds in complex pathways.

Differentiation of mammalian skeletal muscle cells cultured on microcarrier beads in a rotating cell culture system

Science.gov (United States)

Torgan, C. E.; Burge, S. S.; Collinsworth, A. M.; Truskey, G. A.; Kraus, W. E.

2000-01-01

The growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.

Enhancing Nutraceutical Bioavailability from Raw and Cooked Vegetables Using Excipient Emulsions: Influence of Lipid Type on Carotenoid Bioaccessibility from Carrots.

Science.gov (United States)

Zhang, Ruojie; Zhang, Zipei; Zou, Liqiang; Xiao, Hang; Zhang, Guodong; Decker, Eric Andrew; McClements, David Julian

2015-12-09

The influence of the nature of the lipid phase in excipient emulsions on the bioaccessibility and transformation of carotenoid from carrots was investigated using a gastrointestinal tract (GIT) model. Excipient emulsions were fabricated using whey protein as an emulsifier and medium-chain triglycerides (MCT), fish oil, or corn oil as the oil phase. Changes in particle size, charge, and microstructure were measured as the carrot-emulsion mixtures were passed through simulated mouth, stomach, and small intestine regions. Carotenoid bioaccessibility depended on the type of lipids used to form the excipient emulsions (corn oil > fish oil ≫ MCT), which was attributed to differences in the solubilization capacity of mixed micelles formed from different lipid digestion products. The transformation of carotenoids was greater for fish oil and corn oil than for MCT, which may have been due to greater oxidation or isomerization. The bioaccessibility of the carotenoids was higher from boiled than raw carrots, which was attributed to greater disruption of the plant tissue facilitating carotenoid release. In conclusion, excipient emulsions are highly effective at increasing carotenoid bioaccessibility from carrots, but lipid type must be optimized to ensure high efficacy.

Biochar reduces the bioaccumulation of PAHs from soil to carrot (Daucus carota L.) in the rhizosphere: A mechanism study.

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Ni, Ni; Song, Yang; Shi, Renyong; Liu, Zongtang; Bian, Yongrong; Wang, Fang; Yang, Xinglun; Gu, Chenggang; Jiang, Xin

2017-12-01

The aim of this study was to reveal the mechanisms on how biochar reduces bioaccumulation of polycyclic aromatic hydrocarbons (PAHs) in tuberous vegetables. Corn straw-derived biochar pyrolyzed at 300°C (CB300) or bamboo-derived biochar pyrolyzed at 700°C (BB700) was amended into PAH-contaminated soil planted with carrot (Daucus carota L.). After 150days, 2% CB300 or 2% BB700 amendments significantly reduced the bioaccumulation of PAHs in carrot root (pbiochars both showed better effectiveness at reducing the bioavailability of high-molecular-weight PAHs than the low-molecular-weight PAHs in the rhizosphere. Therefore, the mechanisms on how biochar reduces the PAH uptake into carrot are dependent on the type of biochar (e.g., pyrolysis temperature and feedstock) and root presence. Copyright © 2017 Elsevier B.V. All rights reserved.

Oscillating Cell Culture Bioreactor

Science.gov (United States)

Freed, Lisa E.; Cheng, Mingyu; Moretti, Matteo G.

2010-01-01

To better exploit the principles of gas transport and mass transport during the processes of cell seeding of 3D scaffolds and in vitro culture of 3D tissue engineered constructs, the oscillatory cell culture bioreactor provides a flow of cell suspensions and culture media directly through a porous 3D scaffold (during cell seeding) and a 3D construct (during subsequent cultivation) within a highly gas-permeable closed-loop tube. This design is simple, modular, and flexible, and its component parts are easy to assemble and operate, and are inexpensive. Chamber volume can be very low, but can be easily scaled up. This innovation is well suited to work with different biological specimens, particularly with cells having high oxygen requirements and/or shear sensitivity, and different scaffold structures and dimensions. The closed-loop changer is highly gas permeable to allow efficient gas exchange during the cell seeding/culturing process. A porous scaffold, which may be seeded with cells, is fixed by means of a scaffold holder to the chamber wall with scaffold/construct orientation with respect to the chamber determined by the geometry of the scaffold holder. A fluid, with/without biological specimens, is added to the chamber such that all, or most, of the air is displaced (i.e., with or without an enclosed air bubble). Motion is applied to the chamber within a controlled environment (e.g., oscillatory motion within a humidified 37 C incubator). Movement of the chamber induces relative motion of the scaffold/construct with respect to the fluid. In case the fluid is a cell suspension, cells will come into contact with the scaffold and eventually adhere to it. Alternatively, cells can be seeded on scaffolds by gel entrapment prior to bioreactor cultivation. Subsequently, the oscillatory cell culture bioreactor will provide efficient gas exchange (i.e., of oxygen and carbon dioxide, as required for viability of metabolically active cells) and controlled levels of fluid

Carrot, Corn, Lettuce and Soybean Nutrient Contents are ...

Science.gov (United States)

Biochar, the carbon-rich material remaining after pyrolysis of cellulosic and manure feedstocks, has the potential as a soil amendment to sequester carbon and to improve soil water-holding and nutrient properties- thereby enhancing plant growth. However, biochar produced from some feedstocks also could adversely affect crop quality by changing soil pH and reducing nutrients (e.g., Ca, K, Mg, N, Na, and P) in plant tissues. To evaluate effects of biochar on the nutrient quality of four crops, we conducted a greenhouse study using pots with: carrot (Daucus carota cv. Tendersweet), corn (Zea mays, cv. Golden Bantam), lettuce (Lactuca sativa, cv. Black-Seeded Simpson) and soybean (Glycine max cv. Viking 2265). Plants were grown in one of two South Carolina sandy Coastal Plain soils (Norfolk and Coxville Soil Series), along with biochar (1% by weight) produced from pine chips (PC), poultry litter (PL), swine solids (SS), switchgrass (SG), and two blends of pine chips plus poultry litter (PC/PL, 50/50% and 80/20%). Each of the feedstocks and feedstock blends was pyrolyzed at 350, 500, and 700 ̊ C to produce the biochar used to amend the Norfolk and Coxville soils. Effects of biochar on leaf nutrients (% dry weight) statistically varied with species, soil, feedstock and temperature and nutrient. For carrot and lettuce, the PL, PL/PC, and SS biochars generally decreased leaf N, Ca, Mg, and P; while PL and PL/PC increased K and Na. Biochars had little effect on lea

Optimization of the carrot leaf dehydration aiming at the preservation of omega-3 fatty acids

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Vanessa Vivian de Almeida

2009-01-01

Full Text Available The carrot leaf dehydration conditions in air circulation oven were optimized through response surface methodology (RSM for minimizing the degradation of polyunsaturated fatty acids, particularly alpha-linolenic (LNA, 18:3n-3. The optimized leaf drying time and temperature were 43 h and 70 ºC, respectively. The fatty acids (FA were investigated using gas chromatography equipped with a flame ionization detector and fused silica capillary column; FA were identified with standards and based on equivalent-chain-length. LNA and other FA were quantified against C21:0 internal standard. After dehydration, the amount of LNA, quantified in mg/100 g dry matter of dehydrated carrot leaves, were 984 mg.

Isolation and characterization of cDNA clones for carrot extensin and a proline-rich 33-kDa protein

International Nuclear Information System (INIS)

Chen, J.; Varner, J.E.

1985-01-01

Extensins are hydroxyproline-rich glycoproteins associated with most dicotyledonous plant cell walls. To isolate cDNA clones encoding extensin, the authors started by isolating poly(A) + RNA from carrot root tissue, and then translating the RNA in vitro, in the presence of tritiated leucine or proline. A 33-kDa peptide was identified in the translation products as a putative extensin precursor. From a cDNA library constructed with poly(A) + RNA from wounded carrots, one cDNA clone (pDC5) was identified that specifically hybridized to poly(A) + RNA encoding this 33-kDa peptide. They isolated three cDNA clones (pDC11, pDC12, and pDC16) from another cDNA library using pCD5 as a probe. DNA sequence data, RNA hybridization analysis, and hybrid released in vitro translation indicate that the cDNA clones pDC11 encodes extensin and that cDNA clones pDC12 and pDC16 encode the 33-kDa peptide, which as yet has an unknown identity and function. The assumption that the 33-kDa peptide was an extensin precursor was invalid. RNA hybridization analysis showed that RNA encoded by both clone types is accumulated upon wounding

Testicular Sertoli cells influence the proliferation and immunogenicity of co-cultured endothelial cells

International Nuclear Information System (INIS)

Fan, Ping; He, Lan; Pu, Dan; Lv, Xiaohong; Zhou, Wenxu; Sun, Yining; Hu, Nan

2011-01-01

Research highlights: → The proliferation of dramatic increased by co-cultured with Sertoli cells. → VEGF receptor-2 expression of ECs was up-regulated by co-cultured with Sertoli cells. → The MHC expression of ECs induced by INF-γ and IL-6, IL-8 and sICAM induced by TNF-α decreased respectively after co-cultured with Sertoli cells. → ECs co-cultured with Sertoli cells also didn't increase the stimulation index of spleen lymphocytes. -- Abstract: The major problem of the application of endothelial cells (ECs) in transplantation is the lack of proliferation and their immunogenicity. In this study, we co-cultured ECs with Sertoli cells to monitor whether Sertoli cells can influence the proliferation and immunogenicity of co-cultured ECs. Sertoli cells were isolated from adult testicular tissue. ECs were divided into the control group and the experimental group, which included three sub-groups co-cultured with 1 x 10 3 , 1 x 10 4 or 1 x 10 5 cell/ml of Sertoli cells. The growth and proliferation of ECs were observed microscopically, and the expression of vascular endothelial growth factor (VEGF) receptor-2 (KDR) was examined by Western blotting. In another experiment, ECs were divided into the control group, the single culture group and the co-culture group with the optimal concentration of Sertoli cells. After INF-γ and TNF-α were added to the culture medium, MHC II antigen expression was detected by immunofluorescence staining and western blotting; interleukin (IL)-6, IL-8 and soluble intercellular adhesion molecule (sICAM) were measured in the culture medium by ELISA. We demonstrated that 1 x 10 4 cell/ml Sertoli cells promoted the proliferation of co-cultured ECs more dramatically than that in other groups (P 4 cell/ml of the Sertoli cells was most effective in the up-regulation of KDR expression in the co-cultured ECs (P < 0.05). Sertoli cells can effectively suppress INF-γ-induced MHC II antigen expression in co-cultured ECs compared with single

Evaluation of Carrot Pomace (Daucus carota L. as Hypocholesterolemic and Hypolipidemic Agent on Albino Rats

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Abd El-Moneim M.R. AFIFY

2013-02-01

Full Text Available The current study examined the attenuating influence of dietary carrot pomace powder (CaPP on hypercholesterolemia and various oxidative stress-associated with biochemical parameters in hypercholesterolemic rats. Thirty two male albino rats weighing 110±10 g were divided into four groups, the first group received the basal diet only and served as (negative control, the second group received the hypercholesterolemic diet and served as positive control, the other groups received hypercholesterolemic diet supplemented with 10%, 20% CaPP for six weeks. The obtained results revealed that groups supplemented with 10% and 20% CaPP significantly decrease total lipid, total cholesterol, triglycerides, low density lipoprotein cholesterol, liver enzymes: alanine aminotransferase, aspartate aminotransferase compared to positive and negative groups. Organs weight, body weight gain significantly decreased compared with positive control. Moreover dietary carrot pomace powder can used to reduce the body weight and reducing hypercholesterolemic complications. In addition, dietary carrot pomace powder serves to improve the blood picture and to reduce the blood glucose level in hypercholesterolemic rats and could use in obese people for body loss. Data of kidney function (Urea record an increase in CaPP 20% level (26.9±2.96 but this increase was non significant with the negative control group (26.6±3.1.

Long-term culture and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized mesenchymal cells.

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Garba, Abubakar; Acar, Delphine D; Roukaerts, Inge D M; Desmarets, Lowiese M B; Devriendt, Bert; Nauwynck, Hans J

2017-09-01

Mesenchymal cells are multipotent stromal cells with self-renewal, differentiation and immunomodulatory capabilities. We aimed to develop a co-culture model for differentiating hematopoietic cells on top of immortalized mesenchymal cells for studying interactions between hematopoietic and mesenchymal cells, useful for adequately exploring the therapeutic potential of mesenchymal cells. In this study, we investigated the survival, proliferation and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized porcine bone marrow mesenchymal cells for a period of five weeks. Directly after collection, primary porcine bone marrow mesenchymal cells adhered firmly to the bottom of the culture plates and showed a fibroblast-like appearance, one week after isolation. Upon immortalization, porcine bone marrow mesenchymal cells were continuously proliferating. They were positive for simian virus 40 (SV40) large T antigen and the mesenchymal cell markers CD44 and CD55. Isolated red bone marrow cells were added to these immortalized mesenchymal cells. Five weeks post-seeding, 92±6% of the red bone marrow hematopoietic cells were still alive and their number increased 3-fold during five weekly subpassages on top of the immortalized mesenchymal cells. The red bone marrow hematopoietic cells were originally small and round; later, the cells increased in size. Some of them became elongated, while others remained round. Tiny dendrites appeared attaching hematopoietic cells to the underlying immortalized mesenchymal cells. Furthermore, weekly differential-quick staining of the cells indicated the presence of monoblasts, monocytes, macrophages and lymphocytes in the co-cultures. At three weeks of co-culture, flow cytometry analysis showed an increased surface expression of CD172a, CD14, CD163, CD169, CD4 and CD8 up to 37±0.8%, 40±8%, 41±4%, 23±3% and 19±5% of the hematopoietic cells, respectively. In conclusion, continuous mesenchymal cell

The impact of freeze-drying on microstructure and rehydration properties of carrot

NARCIS (Netherlands)

Voda, A.; Homan, N.; Witek, M.; Duijster, A.; Dalen, van G.; Sman, van der R.G.M.; Nijsse, J.; Vliet, van L.J.; As, van H.; Duynhoven, van J.P.M.

2012-01-01

The impact of freeze-drying, blanching and freezing rate pre-treatments on the microstructure and on the rehydration properties of winter carrots were studied by µCT, SEM, MRI and NMR techniques. The freezing rate determines the size of ice crystals being formed that leave pores upon drying. Their

3D Cell Culture in Alginate Hydrogels

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Therese Andersen

2015-03-01

Full Text Available This review compiles information regarding the use of alginate, and in particular alginate hydrogels, in culturing cells in 3D. Knowledge of alginate chemical structure and functionality are shown to be important parameters in design of alginate-based matrices for cell culture. Gel elasticity as well as hydrogel stability can be impacted by the type of alginate used, its concentration, the choice of gelation technique (ionic or covalent, and divalent cation chosen as the gel inducing ion. The use of peptide-coupled alginate can control cell–matrix interactions. Gelation of alginate with concomitant immobilization of cells can take various forms. Droplets or beads have been utilized since the 1980s for immobilizing cells. Newer matrices such as macroporous scaffolds are now entering the 3D cell culture product market. Finally, delayed gelling, injectable, alginate systems show utility in the translation of in vitro cell culture to in vivo tissue engineering applications. Alginate has a history and a future in 3D cell culture. Historically, cells were encapsulated in alginate droplets cross-linked with calcium for the development of artificial organs. Now, several commercial products based on alginate are being used as 3D cell culture systems that also demonstrate the possibility of replacing or regenerating tissue.

Imaging of polarity during zygotic and somatic embryogenesis of carrot (Daucus carota L.)

NARCIS (Netherlands)

Timmers, A.C.J.

1993-01-01

In this thesis a study of the regulation of coordinated growth and the development of polarity during embryogenesis of carrot, Daucus carota L., is described. To this end, several microscopical techniques were used, such as light microscopy, fluorescence microscopy,

Development and preservation of a cocktail beverage produced from Carrot (Daucus carrota), Pineapple (Ananas comosus) and Mango (Mangifera indoca L,)

International Nuclear Information System (INIS)

Boateng, A. A.

2013-07-01

Carrot is known to have the potential of providing vital nutrition as well as health and well-being to consumers. In this study, carrot juice was envisaged as a good vehicle for spreading its nutraceutical benefits. The study was then designed in two parts: (a) Survey to establish the beverage consumption pattern in the Anyaa-Sowutuom District in the Greater Accra Region of Ghana and (b) Carrot juice formulation and optimisation. The survey was conducted by administering a questionnaire to volunteer adult consumers. To develop an acceptable carrot juice beverage, pineapple juice and mango juice were included as components to improve taste and flavour respectively. A constrained, simplex centroid mixture design for three components was employed to optimise the proportions of the components in the final product based on sensory attributes. Chemical analyses as well as shelf stability studies were conducted for the optimised beverage preserved by chemicals and gamma radiation. Data obtained from the survey revealed that juices were very popular among consumers. The choice of a particular beverage was dependent on nutrition, taste and colour. Contour plots generated from sensory data of the products were overlaid to determine the optimum ratios of the components of the cocktail beverage. The optimised product consisted of 50-52% Carrot, 18-20% Pineapple and 28-30% Mango. Analysis of the beverage revealed 223 mg/100ml Potassium, 3.92 mg/100ml Pro-vitamin A (Beta carotene) and 43 mg/100ml vitamin C. After 8 weeks of storage 71% Pro-vitamin A was retained even under relatively higher doses (2.5kGy) of gamma radiation. The microbial quality of the beverage was good under all conditions. (au)

Cell culture experiments planned for the space bioreactor

Science.gov (United States)

Morrison, Dennis R.; Cross, John H.

1987-01-01

Culturing of cells in a pilot-scale bioreactor remains to be done in microgravity. An approach is presented based on several studies of cell culture systems. Previous and current cell culture research in microgravity which is specifically directed towards development of a space bioprocess is described. Cell culture experiments planned for a microgravity sciences mission are described in abstract form.

Advances in cell culture: anchorage dependence

Science.gov (United States)

Merten, Otto-Wilhelm

2015-01-01

Anchorage-dependent cells are of great interest for various biotechnological applications. (i) They represent a formidable production means of viruses for vaccination purposes at very large scales (in 1000–6000 l reactors) using microcarriers, and in the last decade many more novel viral vaccines have been developed using this production technology. (ii) With the advent of stem cells and their use/potential use in clinics for cell therapy and regenerative medicine purposes, the development of novel culture devices and technologies for adherent cells has accelerated greatly with a view to the large-scale expansion of these cells. Presently, the really scalable systems—microcarrier/microcarrier-clump cultures using stirred-tank reactors—for the expansion of stem cells are still in their infancy. Only laboratory scale reactors of maximally 2.5 l working volume have been evaluated because thorough knowledge and basic understanding of critical issues with respect to cell expansion while retaining pluripotency and differentiation potential, and the impact of the culture environment on stem cell fate, etc., are still lacking and require further studies. This article gives an overview on critical issues common to all cell culture systems for adherent cells as well as specifics for different types of stem cells in view of small- and large-scale cell expansion and production processes. PMID:25533097

Exploring the effects of pulsed electric field processing parameters on polyacetylene extraction from carrot slices.

Science.gov (United States)

Aguiló-Aguayo, Ingrid; Abreu, Corina; Hossain, Mohammad B; Altisent, Rosa; Brunton, Nigel; Viñas, Inmaculada; Rai, Dilip K

2015-03-02

The effects of various pulsed electric field (PEF) parameters on the extraction of polyacetylenes from carrot slices were investigated. Optimised conditions with regard to electric field strength (1-4 kV/cm), number of pulses (100-1500), pulse frequency (10-200 Hz) and pulse width (10-30 μs) were identified using response surface methodology (RSM) to maximise the extraction of falcarinol (FaOH), falcarindiol (FaDOH) and falcarindiol-3-acetate (FaDOAc) from carrot slices. Data obtained from RSM and experiments fitted significantly (p pulses of 10 μs at 10 Hz. The predicted values from the developed quadratic polynomial equation were in close agreement with the actual experimental values with low average mean deviations (E%) ranging from 0.68% to 3.58%.

Only the Carrot, Not the Stick: Incorporating Trust into the Enforcement of Regulation

NARCIS (Netherlands)

Mendoza Rodriguez, J.P.; Wielhouwer, J.L.

2015-01-01

New enforcement strategies allow agents to gain the regulator's trust and consequently face a lower audit probability. Prior research suggests that, in order to prevent lower compliance, a reduction in the audit probability (the "carrot ") must be compensated with the introduction of a higher

Biological control of Alternaria radicina in seed production of carrots with Ulocladium atrum

NARCIS (Netherlands)

Köhl, J.; Langerak, C.J.; Meekes, E.T.M.; Molhoek, W.M.L.

2004-01-01

Black rot of carrots is caused by seed-borne Alternaria radicina. Biological control of seed infestation by treatments applied to plants in flower during seed production with the fungal antagonist Ulocladium atrum was investigated in laboratory and field experiments resulting in a reduction of seed

Suspension culture of pluripotent stem cells: effect of shear on stem cell fate.

Science.gov (United States)

Keller, Kevin C; Rodrigues, Beatriz; zur Nieden, Nicole I

2014-01-01

Despite significant promise, the routine usage of suspension cell culture to manufacture stem cell-derived differentiated cells has progressed slowly. Suspension culture is an innovative way of either expanding or differentiating cells and sometimes both are combined into a single bioprocess. Its advantages over static 2D culturing include a homogeneous and controllable culture environment and producing a large quantity of cells in a fraction of time. This feature makes suspension cell culture ideal for use in stem cell research and eventually ideal in the large-scale production of differentiated cells for regenerative medicine. Because of their tremendous differentiation capacities and unlimited growth properties, pluripotent stem cells (PSCs) in particular are considered potential sources for future cell-replacement therapies. Currently, expansion of PSCs is accomplished in 2D, which only permits a limited amount of cell growth per culture flask before cells need to be passaged. However, before stem cells can be applied clinically, several aspects of their expansion, such as directed growth, but also differentiation, need to be better controlled. This review will summarize recent advantages in suspension culture of PSCs, while at the same time highlighting current challenges.

Taxa de respiração de cenouras minimamente processadas e armazenadas em diferentes temperaturas Respiration rate of storage processed carrots at different temperatures

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Wigberto Antonio Spagnol

2006-09-01

Full Text Available Entre as hortaliças minimamente processadas, a cenoura é uma das mais populares, sendo comercializada de várias maneiras: raladas, cortadas em fatias, palitos, e ainda apresentadas na forma de mini-cenoura (baby carrot. O objetivo deste estudo foi determinar as taxas respiratórias de cenouras (Daucus carota da cultivar Nantes minimamente processadas. O armazenamento foi realizado nas temperaturas de 1 °C, 5 °C e 11 °C, e 90% UR. A taxa respiratória foi determinada usando um fluxo contínuo de ar. O teor de CO2 e etileno foi medido por um cromatógrafo a gás. A taxa de respiração para as cenouras fatiadas foi mais alta do que para os produtos inteiros. Os valores da energia de ativação obtidos para as cenouras fatiadas e inteiras foi de 69,82 kJmol-1 e 54,60 kJmol-1, respectivamente. A produção de etileno foi insignificante para as cenouras durante os 14 dias de armazenamento.The carrot is one of the most popular vegetables from minimally processed vegetables. It is commercialized in many different ways: shreds, slices, sticks and baby carrots. The aim of this work is to determine the respiration rate of minimally processed carrots. They were in storage at temperatures of 1 °C, 5 °C and 11 °C, and 90% RH. The respiration rate was determined using continuous humidification airflow and measuring the CO2 concentration using a gas chromatograph connected to a microcomputer. The respiration rates of the minimally processed carrots showed a higher respiration rate than for the whole products. The activation energy values calculated for the minimally processed carrots corresponded to 69.82 kJmol-1 for the whole products. The ethylene production for the carrots remained insignificant throughout the 14 days of storage.

Controlling the diversity of cell populations in a stem cell culture

NARCIS (Netherlands)

Heo, Inha; Clevers, Hans

2015-01-01

Culturing intestinal stem cells into 3D organoids results in heterogeneous cell populations, reflecting the in vivo cell type diversity. In a recent paper published in Nature, Wang et al. established a culture condition for a highly homogeneous population of intestinal stem cells.

Solid-State Fermentation of Carrot Pomace for the Production of Inulinase by Penicillium oxalicum BGPUP-4.

Science.gov (United States)

Singh, Ram Sarup; Chauhan, Kanika; Singh, Jagroop; Pandey, Ashok; Larroche, Christian

2018-03-01

Inulinases are an important class of industrial enzymes which are used for the production of high-fructose syrup and fructooligosaccharides. Inulin, a polyfructan, is generally employed for the production of inulinase, which is a very expensive substrate. A number of agroindustrial residues have been used for cost-effective production of inulinases. In the present study, carrot pomace was selected as a substrate for the production of inulinase by Penicillium oxalicum BGPUP-4 in solid-state fermentation. Carrot pomace is one of the good substrates for bioprocesses, because it is rich in soluble and insoluble carbohydrates. A central composite rotatable design (CCRD) used in response surface methodology was employed for the optimal production of inulinase from carrot pomace. Using CCRD, 15 runs were practiced to optimize the range of three independent variables: moisture content (70-90%), incubation time (4-6 days) and pH (5.0-7.0) for inulinase production. Carrot pomace supplemented with 0.5% inulin as an inducer, 0.2% NH 4 H 2 PO 4 , 0.2% NaNO 3 , 0.2% KH 2 PO 4 , 0.05% MgSO 4 ·7H 2 O and 0.001% FeSO 4 ·7H 2 O was used for the production of inulinase in solid-state fermentation at 30 °C. Inulinase production (322.10 IU per g of dry substrate) was obtained under the optimized conditions, i.e . moisture content of 90%, incubation time 4 days and pH=7.0. The corresponding inulinase/invertase (I/S) ratio (3.38) was also high, which indicates the inulolytic nature of the enzyme. Multiple correlation coefficients R for inulinase production and I/S ratio were 0.9995 and 0.9947, respectively. The R value very close to one indicates an excellent correlation between experimental and predicted results.

The evolution of chicken stem cell culture methods.

Science.gov (United States)

Farzaneh, M; Attari, F; Mozdziak, P E; Khoshnam, S E

2017-12-01

1. The avian embryo is an excellent model for studying embryology and the production of pharmaceutical proteins in transgenic chickens. Furthermore, chicken stem cells have the potential for proliferation and differentiation and emerged as an attractive tool for various cell-based technologies. 2. The objective of these studies is the derivation and culture of these stem cells is the production of transgenic birds for recombinant biomaterials and vaccine manufacture, drug and cytotoxicity testing, as well as to gain insight into basic science, including cell tracking. 3. Despite similarities among the established chicken stem cell lines, fundamental differences have been reported between their culture conditions and applications. Recent conventional protocols used for expansion and culture of chicken stem cells mostly depend on feeder cells, serum-containing media and static culture. 4. Utilising chicken stem cells for generation of cell-based transgenic birds and a variety of vaccines requires large-scale cell production. However, scaling up the conventional adherent chicken stem cells is challenging and labour intensive. Development of a suspension cell culture process for chicken embryonic stem cells (cESCs), chicken primordial germ cells (PGCs) and chicken induced pluripotent stem cells (ciPSCs) will be an important advance for increasing the growth kinetics of these cells. 6. This review describes various approaches and suggestions to achieve optimal cell growth for defined chicken stem cells cultures and use in future manufacturing applications.

Mammalian Cell Culture Simplified.

Science.gov (United States)

Moss, Robert; Solomon, Sondra

1991-01-01

A tissue culture experiment that does not require elaborate equipment and that can be used to teach sterile technique, the principles of animal cell line maintenance, and the concept of cell growth curves is described. The differences between cancerous and normal cells can be highlighted. The procedure is included. (KR)

Rabbit uterine epithelial cells: Co-culture with spermatozoa

International Nuclear Information System (INIS)

Boice, M.L.

1988-01-01

A primary culture of rabbit uterine epithelial cells was established and their effects on sperm function were examined in vitro. Epithelial cells were isolated from uteri of estrous rabbits and cultured on floating collagen gels in phenol red-free medium supplemented with 5% fetal bovine serum. Light microscopy and keratin staining showed that the epithelial cell population established in culture had morphological characteristics similar to that seen in the intact endometrium. Cells were cultured with 3 H-leucine and uptake of label by cells and its incorporation into cellular and secretory proteins determined. When compared to cells cultured for 24-48 h, incorporation of label into cellular protein was lower at 72-96 h, but secretion increased. Estradiol 17-β did not affect label uptake or incorporation, but did enhance proliferation of cells as judged by total DNA content of the cell population. Analysis of proteins in media by sodium dodecyl sulfate polyacrylamide gel electrophoresis and fluorography suggested that epithelial and stromal cells synthesis proteins that may be secretory in nature during 72-96 h culture. Twenty-nine to thirty-one h after initiation of epithelial cultures, 1-2 x 10 6 sperm were co-incubated with cells and sperm viability, motility, loss of acrosome and fertilizing ability determined

Risk Analysis on Organochlorine Pesticides Residue in Potato and Carrot from Conventional and Organic Farms in Citarum Watershed Area, West Java Province, Indonesia

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Suphia Rahmawati

2017-03-01

Full Text Available Six samples of carrot and potatoes were collected from conventional and organic farms. Organochlorine pesticides (OCPs residue was analyzed using liquid-liquid extraction. The concentration of OCPs residue together with the consumption pattern were used to calculate the potential risk. Estimated Daily Intake (EDI of OCPs residue in carrots and potatoes both from conventional and organic farms were below the standard given in SNI 7313:2008 and the Maximum Residue Limit (MRLs from USEPA . Hazard quotient (HQ and Hazard Index (HI as effect analysis were below the unity, thus there may not be a concern for non-carcinogenic effect. Risk analysis on cancer effects indicated that potatoes samples were above the recommended risk by EPA (10-6. In addition, risk of potatoes samples from organic samples was higher compare to that of the conventional farm. Cancer risk analysis of carrot samples were found below the recommended risk by EPA. Cancer risk effect of carrot samples from conventional farm was slightly higher compare to that of the organic farm.

X-ray microanalysis of single and cultured cells

International Nuclear Information System (INIS)

Wroblewski, J.; Roomans, G.M.

1984-01-01

X-ray microanalysis of single or cultured cells is often a useful alternative or complement to the analysis of the corresponding tissue. It also allows the analysis of individual cells in a cell population. Preparation for X-ray microanalysis poses a number of typical problems. Suspensions of single cells can be prepared by either of two pathways: (1) washing - mounting - drying, or (2) centrifugation - freezing or fixation - sectioning. The washing step in the preparation of single or cultured cells presents the most severe problems. Cultured cells are generally grown on a substrate that is compatible with both the analysis and the culture, washed and dried. In some cases, sectioning of cultured cell monolayers has been performed. Special problems in quantitative analysis occur in those cases where the cells are analyzed on a thick substrate, since the substrate contributes to the spectral background

Continued selenium biofortification of carrots and broccoli grown in soils once amended with Se-enriched S. pinnata

Directory of Open Access Journals (Sweden)

Gary S. Bañuelos

2016-08-01

Full Text Available Selenium (Se biofortification has been practiced in Se-deficient regions throughout the world primarily by adding inorganic sources of Se to the soil. Considering the use of adding organic sources of Se could be useful as an alternative Se amendment for the production of Se-biofortified food crops. In this multi-year micro-plot study, we investigate growing carrots and broccoli in soils that had been previously amended with Se-enriched Stanleya pinnata Pursh (Britton three and four years prior to planting one and two, respectively. Results showed that total and extractable Se concentrations in soils (0-30 cm were 1.65 mg kg-1 and 88 µg L-1, and 0.92 mg kg-1 and 48.6 µg L-1 at the beginning of the growing season for planting one and two, respectively. After each respective growing season, total Se concentrations in the broccoli florets and carrots ranged from 6.99 to 7.83 mg kg-1 and 3.15 to 6.25 mg kg-1 in planting one and two, respectively. In broccoli and carrot plant tissues, SeMet (selenomethionine was the predominant selenoamino acid identified in Se aqueous extracts. In postharvest soils from planting one, phospholipid analyses (PLFA showed that amending the soil with S. pinnata exerted no effect on the microbial biomass, AMF (arbuscular mycorrhizal fungi, actinomycetes and Gram-positive and bacterial PLFA at both 0-5 and 0-30 cm, respectively, three years later. Successfully producing Se-enriched broccoli and carrots three and four years later after amending soil with Se-enriched S. pinnata clearly demonstrates its potential source as an organic Se enriched fertilizer for Se-deficient regions.

Effect of simultaneous infrared dry-blanching and dehydration on quality characteristics of carrot slices

Science.gov (United States)

This study investigated the effects of various processing parameters on carrot slices exposed to infrared (IR) radiation heating for achieving simultaneous infrared dry-blanching and dehydration (SIRDBD). The investigated parameters were product surface temperature, slice thickness and processing ti...

The research project KLIMAT. Report of carrot case study; Forskningsprosjektet KLIMAT. Rapport fra gulrot case studie

Energy Technology Data Exchange (ETDEWEB)

Svanes, Erik

2012-07-01

The research project 'KLIMAT' was 1.1.2009 to 31.8.2012. The main goal of this project was to develop a 'best practice' methodology for quantifying the carbon footprint of products with a focus on fruits, vegetables, dairy and meat products. The methodology was tested on six selected case products. One of these products was the carrot. The product was selected because it is a product with high sales volume, a large part of the production takes place in Norway, and it may be assumed to represent the root vegetables which are a very important group of vegetable products. Oestfold Research, in conjunction with BAMA and Gartnerhallen, collected the necessary data to conduct the study. Data came from a group of 13 producers with ties to Laagendalen that all products supplied to the Laagen Gulrot for processing and packing. The producers were voluntarily affiliated with an online reporting system for a variety of information, including information about resource use that were used in this study. Climate track of carrot packed in the cup was, according to the methodology that was developed 0.39 kg Co2-ekv/kg product delivered to the consumer, 0.44 kg Co2-ekv/kg consumed carrot. (Author)

Particle Trajectories in Rotating Wall Cell Culture Devices

Science.gov (United States)

Ramachandran N.; Downey, J. P.

1999-01-01

Cell cultures are extremely important to the medical community since such cultures provide an opportunity to perform research on human tissue without the concerns inherent in experiments on individual humans. Development of cells in cultures has been found to be greatly influenced by the conditions of the culture. Much work has focused on the effect of the motions of cells in the culture relative to the solution. Recently rotating wall vessels have been used with success in achieving improved cellular cultures. Speculation and limited research have focused on the low shear environment and the ability of rotating vessels to keep cells suspended in solution rather than floating or sedimenting as the primary reasons for the improved cellular cultures using these devices. It is widely believed that the cultures obtained using a rotating wall vessel simulates to some degree the effect of microgravity on cultures. It has also been speculated that the microgravity environment may provide the ideal acceleration environment for culturing of cellular tissues due to the nearly negligible levels of sedimentation and shear possible. This work predicts particle trajectories of cells in rotating wall vessels of cylindrical and annular design consistent with the estimated properties of typical cellular cultures. Estimates of the shear encountered by cells in solution and the interactions with walls are studied. Comparisons of potential experiments in ground and microgravity environments are performed.

Usability and Applicability of Microfluidic Cell Culture Systems

DEFF Research Database (Denmark)

Hemmingsen, Mette

possibilities for, for example, precise control of the chemical environment, 3D cultures, controlled co-culture of different cell types or automated, individual control of up to 96 cell culture chambers in one integrated system. Despite the great new opportunities to perform novel experimental designs......Microfluidic cell culture has been a research area with great attention the last decade due to its potential to mimic the in vivo cellular environment more closely compared to what is possible by conventional cell culture methods. Many exciting and complex devices have been presented providing......, these devices still lack general implementation into biological research laboratories. In this project, the usability and applicability of microfluidic cell culture systems have been investigated. The tested systems display good properties regarding optics and compatibility with standard laboratory equipment...

Polyphenolic profile and biological activities of black carrot crude extract (Daucus carota L. ssp. sativus var. atrorubens Alef.).

Science.gov (United States)

Smeriglio, A; Denaro, M; Barreca, D; D'Angelo, V; Germanò, M P; Trombetta, D

2018-01-01

Black carrot (Daucus carota L. ssp. sativus var. atrorubens Alef.) is a valuable source of carbohydrates, minerals and vitamins and contains also high amounts of anthocyanins giving the characteristic deep-purple color. These latter compounds are known as natural dyes used in the food and pharmaceutical industry that have recently attracted much attention for their healthful properties. The aim of this work was to investigate for the first time the polyphenolic profile and biological properties of a black carrot crude extract (BCCE) through an in-depth analysis of the main polyphenolic classes evaluating its antioxidant, cytoprotective and anti-angiogenic properties. Twenty five polyphenols were quantified by LC-DAD-FLD-MS/MS analysis (anthocyanins 78.06%, phenolic acids 17.89% and other flavonoids 4.06%) with polyglycosylated cyanidins as major components. In addition, BCCE showed a strong antioxidant and free radical scavenging activity particularly in the hydrogen transfer-based assays (ORAC and β-carotene bleaching) and a significant increase in the cell viability. Furthermore, BCCE exhibited a strong anti-angiogenic activity at the highest concentration assayed on the chick chorioallantoic membrane (50μg/egg). In conclusion, the obtained results demonstrated the antioxidant, cytoprotective and anti-angiogenic properties of BCCE, which highlight that the higher biological activity of BCCE is probably due to the synergic effects exerted by various polyphenolic classes. Copyright © 2017 Elsevier B.V. All rights reserved.

Good Cell Culture Practice for stem cells and stem-cell-derived models.

Science.gov (United States)

Pamies, David; Bal-Price, Anna; Simeonov, Anton; Tagle, Danilo; Allen, Dave; Gerhold, David; Yin, Dezhong; Pistollato, Francesca; Inutsuka, Takashi; Sullivan, Kristie; Stacey, Glyn; Salem, Harry; Leist, Marcel; Daneshian, Mardas; Vemuri, Mohan C; McFarland, Richard; Coecke, Sandra; Fitzpatrick, Suzanne C; Lakshmipathy, Uma; Mack, Amanda; Wang, Wen Bo; Yamazaki, Daiju; Sekino, Yuko; Kanda, Yasunari; Smirnova, Lena; Hartung, Thomas

2017-01-01

The first guidance on Good Cell Culture Practice (GCCP) dates back to 2005. This document expands this to include aspects of quality assurance for in vitro cell culture focusing on the increasingly diverse cell types and culture formats used in research, product development, testing and manufacture of biotechnology products and cell-based medicines. It provides a set of basic principles of best practice that can be used in training new personnel, reviewing and improving local procedures, and helping to assure standard practices and conditions for the comparison of data between laboratories and experimentation performed at different times. This includes recommendations for the documentation and reporting of culture conditions. It is intended as guidance to facilitate the generation of reliable data from cell culture systems, and is not intended to conflict with local or higher level legislation or regulatory requirements. It may not be possible to meet all recommendations in this guidance for practical, legal or other reasons. However, when it is necessary to divert from the principles of GCCP, the risk of decreasing the quality of work and the safety of laboratory staff should be addressed and any conclusions or alternative approaches justified. This workshop report is considered a first step toward a revised GCCP 2.0.

Effects of inulin, carrot and cellulose fibres on the properties of raw ...

African Journals Online (AJOL)

The effects of inulin, carrot, and cellulose fibres (3%, 6%, and 9%) on raw and fried chicken meatballs were studied. Meatball pH, thiobarbituric acid reactive substances (TBARS), and colour values were determined for raw samples in refrigerated storage on the 1st, 5th and 10th days. The effects of fibres and their various ...

Long-term maintenance of human induced pluripotent stem cells by automated cell culture system.

Science.gov (United States)

Konagaya, Shuhei; Ando, Takeshi; Yamauchi, Toshiaki; Suemori, Hirofumi; Iwata, Hiroo

2015-11-17

Pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem (iPS) cells, are regarded as new sources for cell replacement therapy. These cells can unlimitedly expand under undifferentiated conditions and be differentiated into multiple cell types. Automated culture systems enable the large-scale production of cells. In addition to reducing the time and effort of researchers, an automated culture system improves the reproducibility of cell cultures. In the present study, we newly designed a fully automated cell culture system for human iPS maintenance. Using an automated culture system, hiPS cells maintained their undifferentiated state for 60 days. Automatically prepared hiPS cells had a potency of differentiation into three germ layer cells including dopaminergic neurons and pancreatic cells.

High-Throughput Cancer Cell Sphere Formation for 3D Cell Culture.

Science.gov (United States)

Chen, Yu-Chih; Yoon, Euisik

2017-01-01

Three-dimensional (3D) cell culture is critical in studying cancer pathology and drug response. Though 3D cancer sphere culture can be performed in low-adherent dishes or well plates, the unregulated cell aggregation may skew the results. On contrary, microfluidic 3D culture can allow precise control of cell microenvironments, and provide higher throughput by orders of magnitude. In this chapter, we will look into engineering innovations in a microfluidic platform for high-throughput cancer cell sphere formation and review the implementation methods in detail.

Liver Cell Culture Devices

NARCIS (Netherlands)

Andria, B.; Bracco, A.; Cirino, G.; Chamuleau, R. A. F. M.

2010-01-01

In the last 15 years many different liver cell culture devices, consisting of functional liver cells and artificial materials, have been developed. They have been devised for numerous different applications, such as temporary organ replacement (a bridge to liver transplantation or native liver

Characterization of glucocerebrosidase in peripheral blood cells and cultured blastoid cells

NARCIS (Netherlands)

Aerts, J. M.; Heikoop, J.; van Weely, S.; Donker-Koopman, W. E.; Barranger, J. A.; Tager, J. M.; Schram, A. W.

1988-01-01

We have characterized glucocerebrosidase in various cell types of peripheral blood of control subjects and in cultured human blastoid cells. The intracellular level of glucocerebrosidase in cultured blastoid cells (10-30 nmol substrate hydrolyzed/h.mg protein) resembles closely values observed for

Biosynthesis of 14C-phytoene from tomato cell suspension cultures (Lycopersicon esculentum) for utilization in prostate cancer cell culture studies.

Science.gov (United States)

Campbell, Jessica K; Rogers, Randy B; Lila, Mary Ann; Erdman, John W

2006-02-08

This work describes the development and utilization of a plant cell culture production approach to biosynthesize and radiolabel phytoene and phytofluene for prostate cancer cell culture studies. The herbicide norflurazon was added to established cell suspension cultures of tomato (Lycopersicon esculentum cv. VFNT cherry), to induce the biosynthesis and accumulation of the lycopene precursors, phytoene and phytofluene, in their natural isomeric forms (15-cis-phytoene and two cis-phytofluene isomers). Norflurazon concentrations, solvent carrier type and concentration, and duration of culture exposure to norflurazon were screened to optimize phytoene and phytofluene synthesis. Maximum yields of both phytoene and phytofluene were achieved after 7 days of treatment with 0.03 mg norflurazon/40 mL fresh medium, provided in 0.07% solvent carrier. Introduction of 14C-sucrose to the tomato cell culture medium enabled the production of 14C-labeled phytoene for subsequent prostate tumor cell uptake studies. In DU 145 prostate tumor cells, it was determined that 15-cis-phytoene and an oxidized product of phytoene were taken up and partially metabolized by the cells. The ability to biosynthesize, radiolabel, and isolate these carotenoids from tomato cell cultures is a novel, valuable methodology for further in vitro and in vivo investigations into the roles of phytoene and phytofluene in cancer chemoprevention.

Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices.

Science.gov (United States)

Halldorsson, Skarphedinn; Lucumi, Edinson; Gómez-Sjöberg, Rafael; Fleming, Ronan M T

2015-01-15

Culture of cells using various microfluidic devices is becoming more common within experimental cell biology. At the same time, a technological radiation of microfluidic cell culture device designs is currently in progress. Ultimately, the utility of microfluidic cell culture will be determined by its capacity to permit new insights into cellular function. Especially insights that would otherwise be difficult or impossible to obtain with macroscopic cell culture in traditional polystyrene dishes, flasks or well-plates. Many decades of heuristic optimization have gone into perfecting conventional cell culture devices and protocols. In comparison, even for the most commonly used microfluidic cell culture devices, such as those fabricated from polydimethylsiloxane (PDMS), collective understanding of the differences in cellular behavior between microfluidic and macroscopic culture is still developing. Moving in vitro culture from macroscopic culture to PDMS based devices can come with unforeseen challenges. Changes in device material, surface coating, cell number per unit surface area or per unit media volume may all affect the outcome of otherwise standard protocols. In this review, we outline some of the advantages and challenges that may accompany a transition from macroscopic to microfluidic cell culture. We focus on decisive factors that distinguish macroscopic from microfluidic cell culture to encourage a reconsideration of how macroscopic cell culture principles might apply to microfluidic cell culture. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

Isolation and culture of larval cells from C. elegans.

Directory of Open Access Journals (Sweden)

Sihui Zhang

Full Text Available Cell culture is an essential tool to study cell function. In C. elegans the ability to isolate and culture cells has been limited to embryonically derived cells. However, cells or blastomeres isolated from mixed stage embryos terminally differentiate within 24 hours of culture, thus precluding post-embryonic stage cell culture. We have developed an efficient and technically simple method for large-scale isolation and primary culture of larval-stage cells. We have optimized the treatment to maximize cell number and minimize cell death for each of the four larval stages. We obtained up to 7.8×10(4 cells per microliter of packed larvae, and up to 97% of adherent cells isolated by this method were viable for at least 16 hours. Cultured larval cells showed stage-specific increases in both cell size and multinuclearity and expressed lineage- and cell type-specific reporters. The majority (81% of larval cells isolated by our method were muscle cells that exhibited stage-specific phenotypes. L1 muscle cells developed 1 to 2 wide cytoplasmic processes, while L4 muscle cells developed 4 to 14 processes of various thicknesses. L4 muscle cells developed bands of myosin heavy chain A thick filaments at the cell center and spontaneously contracted ex vivo. Neurons constituted less than 10% of the isolated cells and the majority of neurons developed one or more long, microtubule-rich protrusions that terminated in actin-rich growth cones. In addition to cells such as muscle and neuron that are high abundance in vivo, we were also able to isolate M-lineage cells that constitute less than 0.2% of cells in vivo. Our novel method of cell isolation extends C. elegans cell culture to larval developmental stages, and allows use of the wealth of cell culture tools, such as cell sorting, electrophysiology, co-culture, and high-resolution imaging of subcellular dynamics, in investigation of post-embryonic development and physiology.

Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory.

Science.gov (United States)

Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V McNeil; Segarra, Verónica A

2017-01-01

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented-one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

Multizone Paper Platform for 3D Cell Cultures

Science.gov (United States)

Derda, Ratmir; Hong, Estrella; Mwangi, Martin; Mammoto, Akiko; Ingber, Donald E.; Whitesides, George M.

2011-01-01

In vitro 3D culture is an important model for tissues in vivo. Cells in different locations of 3D tissues are physiologically different, because they are exposed to different concentrations of oxygen, nutrients, and signaling molecules, and to other environmental factors (temperature, mechanical stress, etc). The majority of high-throughput assays based on 3D cultures, however, can only detect the average behavior of cells in the whole 3D construct. Isolation of cells from specific regions of 3D cultures is possible, but relies on low-throughput techniques such as tissue sectioning and micromanipulation. Based on a procedure reported previously (“cells-in-gels-in-paper” or CiGiP), this paper describes a simple method for culture of arrays of thin planar sections of tissues, either alone or stacked to create more complex 3D tissue structures. This procedure starts with sheets of paper patterned with hydrophobic regions that form 96 hydrophilic zones. Serial spotting of cells suspended in extracellular matrix (ECM) gel onto the patterned paper creates an array of 200 micron-thick slabs of ECM gel (supported mechanically by cellulose fibers) containing cells. Stacking the sheets with zones aligned on top of one another assembles 96 3D multilayer constructs. De-stacking the layers of the 3D culture, by peeling apart the sheets of paper, “sections” all 96 cultures at once. It is, thus, simple to isolate 200-micron-thick cell-containing slabs from each 3D culture in the 96-zone array. Because the 3D cultures are assembled from multiple layers, the number of cells plated initially in each layer determines the spatial distribution of cells in the stacked 3D cultures. This capability made it possible to compare the growth of 3D tumor models of different spatial composition, and to examine the migration of cells in these structures. PMID:21573103

Morphology of primary human venous endothelial cell cultures before and after culture medium exchange.

Science.gov (United States)

Krüger-Genge, A; Fuhrmann, R; Jung, F; Franke, R P

2015-01-01

The evaluation of the interaction of human, venous endothelial cells (HUVEC) with body foreign materials on the cellular level cannot be performed in vivo, but is investigated in vitro under standard culture conditions. To maintain the vitality, proliferation and morphology of HUVEC seeded on body foreign substrates over days, the cell culture medium is usually exchanged every second day. It is well known, that alterations in the microenvironment of cells bear the risk of influencing cell morphology and function. In the current study the influence of cell culture medium exchange on HUVEC cytoskeletal microfilament structure and function was investigated. HUVEC in the third passage were seeded on extracellular matrix (ECM) - which was secreted from bovine corneal endothelial cells on glass- until functional confluence was reached. The experiment started 11 days after HUVEC seeding with an exchange of the cell culture medium followed by a staining of the actin microfilaments with phalloidin-rhodamin 1.5 and 5 minutes after medium exchange. The microfilaments were documented by use of an Olympus microscope (IMT-2) equipped with a UV lamp and online connected to a TV chain (Sony XC 50 ST/monochrome) implying an OPTIMAS - Image analysis system. Prostacyclin was analysed in the cell culture supernatant. 1.5 min after culture medium exchange in the functionally confluent cultures a slight disturbance of the actin microfilament structure with a broadening of the marginal filament band, a partial disconnection of cell-cell contacts and the appearance of intercellular fenestrations were observed. 5 minutes after medium exchange a redevelopment of the slightly disturbed microfilament structure with a condensation and narrowing of the marginal filament band was seen. 12 h later a further consolidation of the microfilament structure occurred. In addition, a perturbation of the cultured HUVEC occurred after cell culture medium exchange. The prostacyclin concentration in the

Mutation in cultured mammalian cells

International Nuclear Information System (INIS)

Nakamura, N.; Okada, S.

1982-01-01

Mammalian cell cultures were exposed to gamma-rays at various dose rates. Dose-rate effects were observed in cultured somatic cells of the mouse for cell killing and mutations resistant to 6-thioguanine (TGsup(r)) and to methotrexate (MTXsup(r)). Linear quadratic model may be applied to cell killing and TGsup(r) mutations in some cases but can not explain the whole data. Results at low doses with far low dose-rate were not predictable from data at high doses with acute or chronic irradiation. Radioprotective effects of dimethyl sulfoxide were seen only after acute exposure but not after chronic one, suggesting that damages by indirect action of radiations may be potentially reparable by cells. TGsup(r) mutations seem to contain gross structural changes whereas MTXsup(r) ones may have smaller alterations. (Namekawa, K.)

Cell-free DNA in a three-dimensional spheroid cell culture model

DEFF Research Database (Denmark)

Aucamp, Janine; Calitz, Carlemi; Bronkhorst, Abel J.

2017-01-01

Background Investigating the biological functions of cell-free DNA (cfDNA) is limited by the interference of vast numbers of putative sources and causes of DNA release into circulation. Utilization of three-dimensional (3D) spheroid cell cultures, models with characteristics closer to the in vivo...... cultures can serve as effective, simplified in vivo-simulating “closed-circuit” models since putative sources of cfDNA are limited to only the targeted cells. In addition, cfDNA can also serve as an alternative or auxiliary marker for tracking spheroid growth, development and culture stability. Biological...... significance 3D cell cultures can be used to translate “closed-circuit” in vitro model research into data that is relevant for in vivo studies and clinical applications. In turn, the utilization of cfDNA during 3D culture research can optimize sample collection without affecting the stability of the growth...

Cell culture techniques in honey bee research

Science.gov (United States)

Cell culture techniques are indispensable in most if not all life science disciplines to date. Wherever cell culture models are lacking scientific development is hampered. Unfortunately this has been and still is the case in honey bee research because permanent honey bee cell lines have not yet been...

MEMS-based dynamic cell-to-cell culture platforms using electrochemical surface modifications

International Nuclear Information System (INIS)

Chang, Jiyoung; Lin, Liwei; Yoon, Sang-Hee; Mofrad, Mohammad R K

2011-01-01

MEMS-based biological platforms with the capability of both spatial placements and time releases of living cells for cell-to-cell culture experiments have been designed and demonstrated utilizing electrochemical surface modification effects. The spatial placement is accomplished by electrochemical surface modification of substrate surfaces to be either adhesive or non-adhesive for living cells. The time control is achieved by the electrical activation of the selective indium tin oxide co-culture electrode to allow the migration of living cells onto the electrode to start the cell-to-cell culture studies. Prototype devices have a three-electrode design with an electrode size of 50 × 50 µm 2 and the separation gaps of 2 µm between them. An electrical voltage of −1.5 V has been used to activate the electrodes independently and sequentially to demonstrate the dynamic cell-to-cell culture experiments of NIH 3T3 fibroblast and Madin Darby canine kidney cells. As such, this MEMS platform could be a basic yet versatile tool to characterize transient cell-to-cell interactions

Microfluidic engineered high cell density three-dimensional neural cultures

Science.gov (United States)

Cullen, D. Kacy; Vukasinovic, Jelena; Glezer, Ari; La Placa, Michelle C.

2007-06-01

Three-dimensional (3D) neural cultures with cells distributed throughout a thick, bioactive protein scaffold may better represent neurobiological phenomena than planar correlates lacking matrix support. Neural cells in vivo interact within a complex, multicellular environment with tightly coupled 3D cell-cell/cell-matrix interactions; however, thick 3D neural cultures at cell densities approaching that of brain rapidly decay, presumably due to diffusion limited interstitial mass transport. To address this issue, we have developed a novel perfusion platform that utilizes forced intercellular convection to enhance mass transport. First, we demonstrated that in thick (>500 µm) 3D neural cultures supported by passive diffusion, cell densities =104 cells mm-3), continuous medium perfusion at 2.0-11.0 µL min-1 improved viability compared to non-perfused cultures (p death and matrix degradation. In perfused cultures, survival was dependent on proximity to the perfusion source at 2.00-6.25 µL min-1 (p 90% viability in both neuronal cultures and neuronal-astrocytic co-cultures. This work demonstrates the utility of forced interstitial convection in improving the survival of high cell density 3D engineered neural constructs and may aid in the development of novel tissue-engineered systems reconstituting 3D cell-cell/cell-matrix interactions.

Effect of pulsed electric field treatment on enzyme kinetics and thermostability of endogenous ascorbic acid oxidase in carrots (Daucus carota cv. Nantes).

Science.gov (United States)

Leong, Sze Ying; Oey, Indrawati

2014-03-01

The objective of this research was to study the enzyme kinetics and thermostability of endogenous ascorbic acid oxidase (AAO) in carrot purée (Daucus carota cv. Nantes) after being treated with pulsed electric field (PEF) processing. Various PEF treatments using electric field strength between 0.2 and 1.2kV/cm and pulsed electrical energy between 1 and 520kJ/kg were conducted. The enzyme kinetics and the kinetics of AAO thermal inactivation (55-70°C) were described using Michaelis-Menten model and first order reaction model, respectively. Overall, the estimated Vmax and KM values were situated in the same order of magnitude as the untreated carrot purée after being exposed to pulsed electrical energy between 1 and 400kJ/kg, but slightly changed at pulsed electrical energy above 500kJ/kg. However, AAO presented different thermostability depending on the electric field strength applied. After PEF treatment at the electric field strength between 0.2 and 0.5kV/cm, AAO became thermolabile (i.e. increase in inactivation rate (k value) at reference temperature) but the temperature dependence of k value (Ea value) for AAO inactivation in carrot purée decreased, indicating that the changes in k values were less temperature dependent. It is obvious that PEF treatment affects the temperature stability of endogenous AAO. The changes in enzyme kinetics and thermostability of AAO in carrot purée could be related to the resulting carrot purée composition, alteration in intracellular environment and the effective concentration of AAO released after being subjected to PEF treatment. Copyright © 2013 Elsevier Ltd. All rights reserved.

Antioxidant characterization and sensory evaluation during storage of ultraviolet-B light exposed baby carrots (abstract)

Science.gov (United States)

Baby carrot processing induces wounding stress activation of phenylalanine ammonia-lyase (PAL), enhancing its nutrient content by increasing synthesis of secondary metabolites. Ultraviolet-B (UV-B) exposure further promotes the formation of soluble phenolic compounds, significantly increasing antiox...

Cell sources for in vitro human liver cell culture models

Science.gov (United States)

Freyer, Nora; Damm, Georg; Seehofer, Daniel; Knöspel, Fanny

2016-01-01

In vitro liver cell culture models are gaining increasing importance in pharmacological and toxicological research. The source of cells used is critical for the relevance and the predictive value of such models. Primary human hepatocytes (PHH) are currently considered to be the gold standard for hepatic in vitro culture models, since they directly reflect the specific metabolism and functionality of the human liver; however, the scarcity and difficult logistics of PHH have driven researchers to explore alternative cell sources, including liver cell lines and pluripotent stem cells. Liver cell lines generated from hepatomas or by genetic manipulation are widely used due to their good availability, but they are generally altered in certain metabolic functions. For the past few years, adult and pluripotent stem cells have been attracting increasing attention, due their ability to proliferate and to differentiate into hepatocyte-like cells in vitro. However, controlling the differentiation of these cells is still a challenge. This review gives an overview of the major human cell sources under investigation for in vitro liver cell culture models, including primary human liver cells, liver cell lines, and stem cells. The promises and challenges of different cell types are discussed with a focus on the complex 2D and 3D culture approaches under investigation for improving liver cell functionality in vitro. Finally, the specific application options of individual cell sources in pharmacological research or disease modeling are described. PMID:27385595

Modificações sensoriais em cenoura minimamente processada e armazenada sob refrigeração Sensory modifications of fresh cut carrots stored under refrigeration

Directory of Open Access Journals (Sweden)

Josane Maria Resende

2004-03-01

Full Text Available Foram avaliadas as modificações sensoriais em cenoura minimamente processada em dois tipos de cortes durante o armazenamento sob refrigeração. Semanalmente avaliou-se a aparência, a cor, o aroma, o sabor e a textura de cenouras cv. Nantes, minimamente processadas, acondicionadas em potes de polietileno tereftalato (PET e armazenadas a 7ºC, por 14 dias. Para definir a freqüência de consumo e preferência pelo tipo de corte, amostras do produto foram apresentadas ao público e os resultados foram anotados em porcentagem em relação ao número total de pessoas consultadas. Com o decorrer do armazenamento houve redução nos escores para a aparência e a cor, enquanto que o sabor e a textura melhoraram em ambos os cortes. Cenouras raladas apresentaram melhor cor e aparência em relação à cenoura cortada em rodelas, entretanto o sabor e a textura foram melhores para cenouras em rodelas. As cenouras raladas são as mais indicadas para o consumo uma vez que a aparência é o atributo que mais causa impacto na escolha pelo consumidor e dentro desta a cor é a característica mais relevante. A freqüência de consumo deste tipo de produto esta aumentando gradativamente e em relação à preferência pelo tipo de corte, o consumidor optou pela cenoura ralada pela sua maior praticidade e versatilidade de uso.Sensory modifications were evaluated in carrots cv Nantes lightly processed in two kinds of cut (grated and sliced during the storage under refrigeration. Fresh cut carrots were packed in polyethylene tereftalato (PET boxes and stored at 7ºC for 14 days. The carrot's appearance, color, aroma, flavor and texture were evaluated weekly. To obtain the frequency of consumption and preference of kind of cut, samples were offered to people to try. During the storage of both kinds of cut, a reduction in scores for appearance and color occurred, while flavor and texture scores were improved. Fresh cut carrots in the grated form presented better

Influence of unit operations on the levels of polyacetylenes in minimally processed carrots and parsnips: An industrial trial.

Science.gov (United States)

Koidis, Anastasios; Rawson, Ashish; Tuohy, Maria; Brunton, Nigel

2012-06-01

Carrots and parsnips are often consumed as minimally processed ready-to-eat convenient foods and contain in minor quantities, bioactive aliphatic C17-polyacetylenes (falcarinol, falcarindiol, falcarindiol-3-acetate). Their retention during minimal processing in an industrial trial was evaluated. Carrot and parsnips were prepared in four different forms (disc cutting, baton cutting, cubing and shredding) and samples were taken in every point of their processing line. The unit operations were: peeling, cutting and washing with chlorinated water and also retention during 7days storage was evaluated. The results showed that the initial unit operations (mainly peeling) influence the polyacetylene retention. This was attributed to the high polyacetylene content of their peels. In most cases, when washing was performed after cutting, less retention was observed possibly due to leakage during tissue damage occurred in the cutting step. The relatively high retention during storage indicates high plant matrix stability. Comparing the behaviour of polyacetylenes in the two vegetables during storage, the results showed that they were slightly more retained in parsnips than in carrots. Unit operations and especially abrasive peeling might need further optimisation to make them gentler and minimise bioactive losses. Copyright © 2011 Elsevier Ltd. All rights reserved.

The Effect of Combination Carrot Juice (Daucus carota L. and Hunkwee Flour in Manufacturing Kefir Ice Cream on Physical and Chemical Quality of Kefir Ice Cream

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Ilma Mahdiana

2017-03-01

Full Text Available The purpose of this research was to determine the best combination of carrot juice (CJ and hunkwee flour (HF on manufacturing of kefir ice cream. The method of this research was experiment with Completely Randomized Design, 4 treatments and 4 replication, the treatments were without carrot juice + HF 5% (P0, CJ 1.5% + HF 3.5% (P1, CJ 3% + HF 2% (P2, CJ 4.5% + HF 0.5% (P3, the presentage based on Ice Cream Mix (ICM. The variables measured were antioxidant activity, viscosity, total solid and organoleptic (textur, taste and aroma. The data was analized by using Analysis Of Variance (ANOVA continued by Honestly Significance Difference (HSD test. The result of this research showed that the combination of carrot juice and hunkwee flour gave highly significant difference effect (P0.05 on aroma. Conclusion: the combination of carrot juice 1.5% + hunkwee flour 3.5% (P1 in kefir ice cream gave the best result.

Growth of melanocytes in human epidermal cell cultures

International Nuclear Information System (INIS)

Staiano-Coico, L.; Hefton, J.M.; Amadeo, C.; Pagan-Charry, I.; Madden, M.R.; Cardon-Cardo, C.

1990-01-01

Epidermal cell cultures were grown in keratinocyte-conditioned medium for use as burn wound grafts; the melanocyte composition of the grafts was studied under a variety of conditions. Melanocytes were identified by immunohistochemistry based on a monoclonal antibody (MEL-5) that has previously been shown to react specifically with melanocytes. During the first 7 days of growth in primary culture, the total number of melanocytes in the epidermal cultures decreased to 10% of the number present in normal skin. Beginning on day 2 of culture, bipolar melanocytes were present at a mean cell density of 116 +/- 2/mm2; the keratinocyte to melanocyte ratio was preserved during further primary culture and through three subpassages. Moreover, exposure of cultures to mild UVB irradiation stimulated the melanocytes to proliferate, suggesting that the melanocytes growing in culture maintained their responsiveness to external stimuli. When the sheets of cultured cells were enzymatically detached from the plastic culture flasks before grafting, melanocytes remained in the basal layer of cells as part of the graft applied to the patient

Cell Culture as an Alternative in Education.

Science.gov (United States)

Nardone, Roland M.

1990-01-01

Programs that are intended to inform and provide "hands-on" experience for students and to facilitate the introduction of cell culture-based laboratory exercises into the high school and college laboratory are examined. The components of the CellServ Program and the Cell Culture Toxicology Training Programs are described. (KR)

The usefulness of three-dimensional cell culture in induction of cancer stem cells from esophageal squamous cell carcinoma cell lines

International Nuclear Information System (INIS)

Fujiwara, Daisuke; Kato, Kazunori; Nohara, Shigeo; Iwanuma, Yoshimi; Kajiyama, Yoshiaki

2013-01-01

Highlights: •Spheroids were created from esophageal carcinoma cells using NanoCulture® Plates. •The proportion of strongly ALDH-positive cells increased in 3-D culture. •Expression of cancer stem cell-related genes was enhanced in 3-D culture. •CA-9 expression was enhanced, suggesting hypoxia had been induced in 3-D culture. •Drug resistance was increased. 3-D culture is useful for inducing cancer stem cells. -- Abstract: In recent years, research on resistance to chemotherapy and radiotherapy in cancer treatment has come under the spotlight, and researchers have also begun investigating the relationship between resistance and cancer stem cells. Cancer stem cells are assumed to be present in esophageal cancer, but experimental methods for identification and culture of these cells have not yet been established. To solve this problem, we created spheroids using a NanoCulture® Plate (NCP) for 3-dimensional (3-D) cell culture, which was designed as a means for experimentally reproducing the 3-D structures found in the body. We investigated the potential for induction of cancer stem cells from esophageal cancer cells. Using flow cytometry we analyzed the expression of surface antigen markers CD44, CD133, CD338 (ABCG2), CD318 (CDCP1), and CD326 (EpCAM), which are known cancer stem cell markers. None of these surface antigen markers showed enhanced expression in 3-D cultured cells. We then analyzed aldehyde dehydrogenase (ALDH) enzymatic activity using the ALDEFLUOR reagent, which can identify immature cells such as stem cells and precursor cells. 3-D-cultured cells were strongly positive for ALDH enzyme activity. We also analyzed the expression of the stem cell-related genes Sox-2, Nanog, Oct3/4, and Lin28 using RT-PCR. Expression of Sox-2, Nanog, and Lin28 was enhanced. Analysis of expression of the hypoxic surface antigen marker carbonic anhydrase-9 (CA-9), which is an indicator of cancer stem cell induction and maintenance, revealed that CA-9 expression

PECULIARITIES OF SECONDARY METABOLITES BIOSYNTHESIS IN PLANT CELL CULTURES

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A.M. NOSOV

2014-06-01

Full Text Available metabolites formation in plant cell cultures of Panax spp., (ginsenosides; Dioscorea deltoidea (steroid glycosides; Ajuga reptans, Serratula coronata, Rhaponticum carthamoides (ecdisteroids; Polyscias spp., (triterpene glycosides, Taxus spp. (taxoids, Stevia rebaudiana (diterpene steviol-glycosides, Stephania glabra (alkaloids. They are some regular trends of secondary metabolites synthesis in the plant cell culture:It can be noted the stable synthesis of the compound promoting cell proliferation. Indeed, cell cultures of Dioscorea deltoidea were demonstrated to accumulate only furostanol glycosides, which promoted cell division. Furostanol glycoside content of Dioscorea strain DM-0.5 was up to 6 - 12% by dry biomass.Panax ginseng and P. japonicus plant cell cultures synthesize as minimum seven triterpene glycosides (ginsenosides, the productivity of these compounds was up to 6.0 - 8.0% on dry biomass.By contrast, the detectable synthesis of diterpene steviol-glycosides in cultivated cells of Stevia rebaudiana initiated in the mixotrophic cultures during chloroplast formation only.Despite these differences, or mainly due to them, plant cell cultures have become an attractive source of phytochemicals in alternative to collecting wild plants. It provides a guideline to bioreactor-based production of isoprenoids using undifferentiated plant cell cultures. 

Substrate utilisation by plant-cell cultures

Energy Technology Data Exchange (ETDEWEB)

Fowler, M W

1982-01-01

Plant cell cultures have been grown on a wide range of carbon sources in addition to the traditional ones of sucrose and glucose. Biomass yields and growth rates vary greatly between the different carbon sources and there is a variation in response between different cell cultures to individual carbon sources. Some attempts have been made to grow cell cultures on 'waste' and related carbon sources, such as lactose, maltose, starch, molasses and milk whey. Only maltose was found to support growth to anything near the levels observed with glucose and sucrose. In the case of molasses carbon source cell growth was either non-existent or only just measurable. All the data point to glucose as being the most suitable carbon source, principally on the grounds of biomass yield and growth rate. It should be noted, however, that other carbon sources do appear to have a major (positive) influence on natural product synthesis. Uptake into the cell is an important aspect of carbohydrate utilisation. There is strong evidence that from disaccharides upwards, major degradation to smaller units occurs before uptake. In some cases the necessary enzymes appear to be excreted into the culture broth, in others they may be located within the cell wall; invertase that hydrolyses sucrose is a good example. Once the products of carbohydrate degradation and mobilisation enter the cell they may suffer one of two fates, oxidation or utilisation for biosynthesis. The precise split between these two varies depending on such factors as cell growth rate, cell size, nutrient broth composition and carbohydrate status of the cells. In general rapidly growing cells have a high rate of oxidation, whereas cells growing more slowly tend to be more directed towards biosynthesis. Carbohydrate utilisation is a key area of study, underpinning as it does both biomass yield and natural product synthesis. (Refs. 13).

Biona-C Cell Culture pH Monitoring System

Science.gov (United States)

Friedericks, C.

1999-01-01

Sensors 2000! is developing a system to demonstrate the ability to perform accurate, real-time measurements of pH and CO2 in a cell culture media in Space. The BIONA-C Cell Culture pH Monitoring System consists of S2K! developed ion selective sensors and control electronics integrated with the fluidics of a cell culture system. The integrated system comprises a "rail" in the Cell Culture Module (CCM) of WRAIR (Space Biosciences of Walter Read Army Institute of Research). The CCM is a Space Shuttle mid-deck locker experiment payload. The BIONA-C is displayed along with associated graphics and text explanations. The presentation will stimulate interest in development of sensor technology for real-time cell culture measurements. The transfer of this technology to other applications will also be of interest. Additional information is contained in the original document.

Effects of coating on the acceptability of baby-carrots / Efeito do revestimento na aceitabilidade de mini-cenouras

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Fábio Yamashita

2008-08-01

Full Text Available The consumer’s opinion of a product is influenced by its appearance. This, in turn, will influence the decision to purchase and eat the product. Equally, the shelf life and acceptability of carrots subjected to minimal processing is influenced by the fading or superficial blanching that develops in the product. The aim of this work was to evaluate the sensorial acceptability of mini-carrots in relation to the colour. colour. Carrots of the variety Indiana-F1 were processed in the premises of ‘ Mr. Rabbit Farm Ltd ‘ and divided in two lots, where one followed the normal flow of processing for the industry and to the other a coating stage was added, using an aqueous 2% solution of polypeptides. The two lots were stored in upright racks, using two temperatures, 5ºC and 10ºC, for subsequent sensorial evaluation. The acceptability tests of the 4 samples of baby-carrots were conducted after 1, 5, 9, 13, 17, 21 and 25 days of storage. Each time, from 100 to 125 consumers evaluated the baby-carrots using a 9 point hedonic scale. The evaluations were accomplished in three existing supermarkets in Viçosa. After one day of storage, the spacial separation of the samples suggested the existence of three groups, one formed by samples of baby-carrots without coating at 5º C and coated at 10º C, and the other two comprised baby-carrots coated at 5º C and not coated at 10 ºC. The consumers’ distribution demonstrated that the acceptance was quite homogeneous, indicating there was no definite preference of the consumers for any specific sample. On the 5th to the 25th day, the spacial separation of the samples suggested there were two different groups with relation to their acceptability, one for samples of coated baby-carrots and the other for non-coated, independently of storage temperature. The group formed by samples of coated babycarrots showed better acceptance to a larger number of consumers. The results indicated the efficiency of the coating

Home-based preparation approaches altered the availability of health beneficial components from carrots and blueberries

Science.gov (United States)

This study investigated the effects of different home food preparation methods on the availability of the total phenolic contents (TPC) and radical scavenging components, as well as the selected health beneficial compounds from fresh blueberries and carrots. High performance liquid chromatography (...

Rotary orbital suspension culture of embryonic stem cell-derived neural stem/progenitor cells: impact of hydrodynamic culture on aggregate yield, morphology and cell phenotype.

Science.gov (United States)

Laundos, Tiago L; Silva, Joana; Assunção, Marisa; Quelhas, Pedro; Monteiro, Cátia; Oliveira, Carla; Oliveira, Maria J; Pêgo, Ana P; Amaral, Isabel F

2017-08-01

Embryonic stem (ES)-derived neural stem/progenitor cells (ES-NSPCs) constitute a promising cell source for application in cell therapies for the treatment of central nervous system disorders. In this study, a rotary orbital hydrodynamic culture system was applied to single-cell suspensions of ES-NSPCs, to obtain homogeneously-sized ES-NSPC cellular aggregates (neurospheres). Hydrodynamic culture allowed the formation of ES-NSPC neurospheres with a narrower size distribution than statically cultured neurospheres, increasing orbital speeds leading to smaller-sized neurospheres and higher neurosphere yield. Neurospheres formed under hydrodynamic conditions (72 h at 55 rpm) showed higher cell compaction and comparable percentages of viable, dead, apoptotic and proliferative cells. Further characterization of cellular aggregates provided new insights into the effect of hydrodynamic shear on ES-NSPC behaviour. Rotary neurospheres exhibited reduced protein levels of N-cadherin and β-catenin, and higher deposition of laminin (without impacting fibronectin deposition), matrix metalloproteinase-2 (MMP-2) activity and percentage of neuronal cells. In line with the increased MMP-2 activity levels found, hydrodynamically-cultured neurospheres showed higher outward migration on laminin. Moreover, when cultured in a 3D fibrin hydrogel, rotary neurospheres generated an increased percentage of neuronal cells. In conclusion, the application of a constant orbital speed to single-cell suspensions of ES-NSPCs, besides allowing the formation of homogeneously-sized neurospheres, promoted ES-NSPC differentiation and outward migration, possibly by influencing the expression of cell-cell adhesion molecules and the secretion of proteases/extracellular matrix proteins. These findings are important when establishing the culture conditions needed to obtain uniformly-sized ES-NSPC aggregates, either for use in regenerative therapies or in in vitro platforms for biomaterial development or

Protein biosynthesis in cultured human hair follicle cells.

Science.gov (United States)

Weterings, P J; Vermorken, A J; Bloemendal, H

1980-10-31

A new technique has been used for culturing human keratinocytes. The cells grow on the basement membrane-like capsules of bovine lenses. Lens cells were removed from the capsules by rigid trypsinization. In order to exclude any contamination with remaining living cells the isolated capsules were irradiated with X-rays at a dose of 10,000 rad. In this way human epithelial cells can be brought in culture from individual hair follicles. Since feeder cells are not used in this culture technique, the biosynthesis of keratinocyte proteins can be studied in these cultures. The newly synthesized proteins can be separated into a water-soluble, a urea-soluble, and a urea-insoluble fraction. Product analysis has been performed on the first two fractions revealing protein patterns identical to those of intact hair follicles. Product analysis of the urea-soluble fractions of microdissected hair follicles shows that the protein pattern of the cultured keratinocytes resembles the protein pattern of the hair follicle sheath. Studies on the metabolism of benzo(a)pyrene revealed that the enzyme aryl hydrocarbon hydroxylase (AHH) is present in cultured hair follicle cells. A possible use of our culture system for eventual detection of inherited predisposition for smoking-dependent lung cancer is discussed.

21 CFR 864.2280 - Cultured animal and human cells.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Cultured animal and human cells. 864.2280 Section... Cultured animal and human cells. (a) Identification. Cultured animal and human cells are in vitro cultivated cell lines from the tissue of humans or other animals which are used in various diagnostic...

Culture of human mesenchymal stem cells using a candidate pharmaceutical grade xeno-free cell culture supplement derived from industrial human plasma pools.

Science.gov (United States)

Díez, José M; Bauman, Ewa; Gajardo, Rodrigo; Jorquera, Juan I

2015-03-13

Fetal bovine serum (FBS) is an animal product used as a medium supplement. The animal origin of FBS is a concern if cultured stem cells are to be utilized for human cell therapy. Therefore, a substitute for FBS is desirable. In this study, an industrial, xeno-free, pharmaceutical-grade supplement for cell culture (SCC) under development at Grifols was tested for growth of human mesenchymal stem cells (hMSCs), cell characterization, and differentiation capacity. SCC is a freeze-dried product obtained through cold-ethanol fractionation of industrial human plasma pools from healthy donors. Bone marrow-derived hMSC cell lines were obtained from two commercial suppliers. Cell growth was evaluated by culturing hMSCs with commercial media or media supplemented with SCC or FBS. Cell viability and cell yield were assessed with an automated cell counter. Cell surface markers were studied by indirect immunofluorescence assay. Cells were cultured then differentiated into adipocytes, chondrocytes, osteoblasts, and neurons, as assessed by specific staining and microscopy observation. SCC supported the growth of commercial hMSCs. Starting from the same number of seeded cells in two consecutive passages of culture with medium supplemented with SCC, hMSC yield and cell population doubling time were equivalent to the values obtained with the commercial medium and was consistent among lots. The viability of hMSCs was higher than 90%, while maintaining the characteristic phenotype of undifferentiated hMSCs (positive for CD29, CD44, CD90, CD105, CD146, CD166 and Stro-1; negative for CD14 and CD19). Cultured hMSCs maintained the potential for differentiation into adipocytes, chondrocytes, osteoblasts, and neurons. The tested human plasma-derived SCC sustains the adequate growth of hMSCs, while preserving their differentiation capacity. SCC can be a potential candidate for cell culture supplement in advanced cell therapies.

Differential marker expression by cultures rich in mesenchymal stem cells

Science.gov (United States)

2013-01-01

Background Mesenchymal stem cells have properties that make them amenable to therapeutic use. However, the acceptance of mesenchymal stem cells in clinical practice requires standardized techniques for their specific isolation. To date, there are no conclusive marker (s) for the exclusive isolation of mesenchymal stem cells. Our aim was to identify markers differentially expressed between mesenchymal stem cell and non-stem cell mesenchymal cell cultures. We compared and contrasted the phenotype of tissue cultures in which mesenchymal stem cells are rich and rare. By initially assessing mesenchymal stem cell differentiation, we established that bone marrow and breast adipose cultures are rich in mesenchymal stem cells while, in our hands, foreskin fibroblast and olfactory tissue cultures contain rare mesenchymal stem cells. In particular, olfactory tissue cells represent non-stem cell mesenchymal cells. Subsequently, the phenotype of the tissue cultures were thoroughly assessed using immuno-fluorescence, flow-cytometry, proteomics, antibody arrays and qPCR. Results Our analysis revealed that all tissue cultures, regardless of differentiation potential, demonstrated remarkably similar phenotypes. Importantly, it was also observed that common mesenchymal stem cell markers, and fibroblast-associated markers, do not discriminate between mesenchymal stem cell and non-stem cell mesenchymal cell cultures. Examination and comparison of the phenotypes of mesenchymal stem cell and non-stem cell mesenchymal cell cultures revealed three differentially expressed markers – CD24, CD108 and CD40. Conclusion We indicate the importance of establishing differential marker expression between mesenchymal stem cells and non-stem cell mesenchymal cells in order to determine stem cell specific markers. PMID:24304471

Chromosomal instability and telomere shortening in long-term culture of hematopoietic stem cells: insights from a cell culture model of RPS14 haploinsufficiency.

Science.gov (United States)

Thomay, K; Schienke, A; Vajen, B; Modlich, U; Schambach, A; Hofmann, W; Schlegelberger, B; Göhring, G

2014-01-01

The fate of cultivated primary hematopoietic stem cells (HSCs) with respect to genetic instability and telomere attrition has not yet been described in great detail. Thus, knowledge of the genetic constitution of HSCs is important when interpreting results of HSCs in culture. While establishing a cell culture model for myelodysplastic syndrome with a deletion in 5q by performing RPS14 knockdown, we found surprising data that may be of importance for any CD34+ cell culture experiments. We performed cytogenetic analyses and telomere length measurement on transduced CD34+ cells and untransduced control cells to observe the effects of long-term culturing. Initially, CD34+ cells had a normal median telomere length of about 12 kb and showed no signs of chromosomal instability. During follow-up, the median telomere length seemed to decrease and, simultaneously, increased chromosomal instability could be observed - in modified and control cells. One culture showed a clonal monosomy 7 - independent of prior RPS14 knockdown. During further culturing, it seemed that the telomeres re-elongated, and chromosomes stabilized, while TERT expression was not elevated. In summary, irrespective of our results of RPS14 knockdown in the long-term culture of CD34+ cells, it becomes clear that cell culture artefacts inducing telomere shortening and chromosomal instability have to be taken into account and regular cytogenetic analyses should always be performed. © 2013 S. Karger AG, Basel.

Supplementation of Carrot (Daucus carrota as Natural Source of Beta-carotene prepared by Several Methods in Feed to Increase Blue Colour of Freshwater Crayfish Red Claw (Cherax quadricarinatus

Directory of Open Access Journals (Sweden)

Woro H. Satyantini

2012-02-01

Full Text Available Red claw (Cherax quadricarinatus is one kind of freshwater crayfish that can cultured as consumption or ornamental fish commodities.  The colour of body was important character for performance of red claw as ornamental fish commodity.  Red claw with blue body colour is more expensive than brown.  The purpose of  this experiment was to know the influence addition of carrot with several methods in feed on blue colour  of freshwater crayfish red claw and which was the best preparation method to increase blue colour of freshwater crayfish red claw. The experiment used Completely Randomized Design with four treatments: A (control, B (extract, C (juice and D (shreeded of carrot with 45 ppm dose and five time replicates.  Data was analysed with Kruskal-Wallis and continued with Z test. Result of the experiment was showed that addition of carrot as natural source of beta-carotene with several preparation in feed increase significantly (P Keywords: freshwater crayfish, colour of red claw, carrot, beta-carotene   ABSTRAK Red claw (Cherax quadricarinatus adalah salah satu jenis lobster air tawar yang dapat dibudidayakan sebagai komoditas konsumsi maupun hias.  Sebagai komoditas ikan hias, warna tubuh merupakan karakter penting untuk menunjang penampilan red claw.  Red claw ukuran konsumsi yang memiliki warna biru akan memiliki nilai jual yang lebih tinggi daripada red claw berwarna coklat.  Tujuan penelitian ini untuk mengetahui pengaruh penambahan wortel sebagai sumber beta karoten alami dengan beberapa metoda pengolahan pada pakan terhadap peningkatan warna biru lobster red claw. Penelitian menggunakan Rancangan Acak Lengkap dengan 4 perlakuan : A (kontrol, B (ekstrak, C (jus dan D (serutan wortel dengan menggunakan dosis 45 ppm beta karoten dan ulangan 5 kali.  Data dianalisis dengan uji Kruskal-Wallis dan dilanjutkan dengan uji Z.  Hasil penelitian menunjukkan bahwa penambahan wortel sebagai sumber beta karoten alami dengan beberapa metode

A single-cell and feeder-free culture system for monkey embryonic stem cells.

Science.gov (United States)

Ono, Takashi; Suzuki, Yutaka; Kato, Yosuke; Fujita, Risako; Araki, Toshihiro; Yamashita, Tomoko; Kato, Hidemasa; Torii, Ryuzo; Sato, Naoya

2014-01-01

Primate pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), hold great potential for research and application in regenerative medicine and drug discovery. To maximize primate PSC potential, a practical system is required for generating desired functional cells and reproducible differentiation techniques. Much progress regarding their culture systems has been reported to date; however, better methods would still be required for their practical use, particularly in industrial and clinical fields. Here we report a new single-cell and feeder-free culture system for primate PSCs, the key feature of which is an originally formulated serum-free medium containing FGF and activin. In this culture system, cynomolgus monkey ESCs can be passaged many times by single-cell dissociation with traditional trypsin treatment and can be propagated with a high proliferation rate as a monolayer without any feeder cells; further, typical PSC properties and genomic stability can be retained. In addition, it has been demonstrated that monkey ESCs maintained in the culture system can be used for various experiments such as in vitro differentiation and gene manipulation. Thus, compared with the conventional culture system, monkey ESCs grown in the aforementioned culture system can serve as a cell source with the following practical advantages: simple, stable, and easy cell maintenance; gene manipulation; cryopreservation; and desired differentiation. We propose that this culture system can serve as a reliable platform to prepare primate PSCs useful for future research and application.

Stimulation of the proliferation of hemopoietic stem cells in irradiated bone marrow cell culture

International Nuclear Information System (INIS)

Mori, K.J.; Izumi, H.; Seto, A.

1981-01-01

Long-term hemopoiesis was established in bone marrow cell culture in vitro. This culture was shown to support the recovery proliferation of hemopoietic stem cells completely in vitro after irradiation. Hemopoietic stem cells were stimulated into proliferation in culture when normal bone marrow cells were overlayed on top of the irradiated adherent cell colonies. These results indicate that proliferation and differentiation of hemopoietic stem cells in vitro are also supported by stromahemopoietic cell interactions

Combination treatment of alkaline electrolyzed water and citric acid with mild heat to ensure microbial safety, shelf-life and sensory quality of shredded carrots.

Science.gov (United States)

Rahman, S M E; Jin, Yong-Guo; Oh, Deog-Hwan

2011-05-01

The objective of this study was to determine the synergistic effect of alkaline electrolyzed water and citric acid with mild heat against background and pathogenic microorganisms on carrots. Shredded carrots were inoculated with approximately 6-7 log CFU/g of Escherichia coli O157:H7 (932, and 933) and Listeria monocytogenes (ATCC 19116, and 19111) and then dip treated with alkaline electrolyzed water (AlEW), acidic electrolyzed water (AcEW), 100 ppm sodium hypochlorite (NaOCl), deionized water (DaIW), or 1% citric acid (CA) alone or with combinations of AlEW and 1% CA (AlEW + CA). The populations of spoilage bacteria on the carrots were investigated after various exposure times (1, 3, and 5 min) and treatment at different dipping temperatures (1, 20, 40, and 50 °C) and then optimal condition (3 min at 50 °C) was applied against foodborne pathogens on the carrots. When compared to the untreated control, treatment AcEW most effectively reduced the numbers of total bacteria, yeast and fungi, followed by AlEW and 100 ppm NaOCl. Exposure to all treatments for 3 min significantly reduced the numbers of total bacteria, yeast and fungi on the carrots. As the dipping temperature increased from 1 °C to 50 °C, the reductions of total bacteria, yeast and fungi increased significantly from 0.22 to 2.67 log CFU/g during the wash treatment (p ≤ 0.05). The combined 1% citric acid and AlEW treatment at 50 °C showed a reduction of the total bacterial count and the yeast and fungi of around 3.7 log CFU/g, as well as effective reduction of L. monocytogenes (3.97 log CFU/g), and E. Coli O157:H7 (4 log CFU/g). Combinations of alkaline electrolyzed water and citric acid better maintained the sensory and microbial quality of the fresh-cut carrots and enhanced the overall shelf-life of the produce. Copyright © 2010 Elsevier Ltd. All rights reserved.

nfluences of ammonium-nitrate, food waste compost and bacterial fertilizer on soluble soil nitrogen forms and on the growth of carrot (Daucus Carota L.

Directory of Open Access Journals (Sweden)

Andrea Balla Kovács

2014-04-01

Full Text Available This paper reports a greenhouse study to compare the effects of food waste compost, bacterial fertilizer and their combination with the effect of mineral fertilizer on yield of carrot and the available nutrient content of soils. The study was conducted on calcareous chernozem and acidic sandy soils and consisted of 8 treatments in a randomized complete block design with four replications. The NH4NO3 resulted in reduced growing of carrot plant in sandy soil, and the treatment effect of mineral fertilizer was not observed significantly in chernozem soil. Sandy soil showed higher response of growth of carrot to food waste compost fertilization than chernozem soil. Sole application of EM-1 bacterial fertilizer did not have marked effect on yield parameters and sizes of roots. When EM-1 bacterial fertilizer was applied together with ammonium-nitrate or with compost in chernozem soil, the weights of roots and the sizes of roots in some cases became higher compared to the values of appropriate treatments without inoculation. In sandy soil the diameter of roots slightly increased when EM-1 bacterial fertilizer was applied with ammonium-nitrate and with ammonium-nitrate+compost combination compared to appropriate treatment without inoculation. In chernozem soil the maximum weights and sizes of roots were achieved with the combined treatment of ammonium-nitrate+compost+EM-1 bacterial fertilizer and in sandy soil with compost treatment. Our results of soluble nitrogen content of soils are in good agreement with yield parameters of carrot. Results suggest that food waste compost could be a good substitute for mineral fertilizer application in carrot production mainly in sandy soil. EM-1 bacterial fertilizer did not cause marked effect on yield and yield parameters of carrot plant, but its combination with other fertilizers promises a little bit higher yield or plant available nutrient in the soil. These effects do not clear exactly, so further studies are

Cell Culture in Microgravity: Opening the Door to Space Cell Biology

Science.gov (United States)

Pellis, Neal R.; Dawson, David L. (Technical Monitor)

1999-01-01

Adaptational response of human cell populations to microgravity is investigated using simulation, short-term Shuttle experiments, and long-term microgravity. Simulation consists of a clinostatically-rotated cell culture system. The system is a horizontally-rotated cylinder completely filled with culture medium. Low speed rotation results in continuous-fall of the cells through the fluid medium. In this setting, cells: 1) aggregate, 2) propagate in three dimensions, 3) synthesize matrix, 4) differentiate, and 5) form sinusoids that facilitate mass transfer. Space cell culture is conducted in flight bioreactors and in static incubators. Cells grown in microgravity are: bovine cartilage, promyelocytic leukemia, kidney proximal tubule cells, adrenal medulla, breast and colon cancer, and endothelium. Cells were cultured in space to test specific hypotheses. Cartilage cells were used to determine structural differences in cartilage grown in space compared to ground-based bioreactors. Results from a 130-day experiment on Mir revealed that cartilage grown in space was substantially more compressible due to insufficient glycosaminoglycan in the matrix. Interestingly, earth-grown cartilage conformed better to the dimensions of the scaffolding material, while the Mir specimens were spherical. The other cell populations are currently being analyzed for cell surface properties, gene expression, and differentiation. Results suggest that some cells spontaneously differentiate in microgravity. Additionally, vast changes in gene expression may occur in response to microgravity. In conclusion, the transition to microgravity may constitute a physical perturbation in cells resulting in unique gene expressions, the consequences of which may be useful in tissue engineering, disease modeling, and space cell biology.

How do culture media influence in vitro perivascular cell behavior?

Science.gov (United States)

Huber, Birgit; Volz, Ann-Cathrin; Kluger, Petra Juliane

2015-12-01

Perivascular cells are multilineage cells located around the vessel wall and important for wall stabilization. In this study, we evaluated a stem cell media and a perivascular cell-specific media for the culture of primary perivascular cells regarding their cell morphology, doubling time, stem cell properties, and expression of cell type-specific markers. When the two cell culture media were compared to each other, perivascular cells cultured in the stem cell medium had a more elongated morphology and a faster doubling rate and cells cultured in the pericyte medium had a more typical morphology, with several filopodia, and a slower doubling rate. To evaluate stem cell properties, perivascular cells, CD146(-) cells, and mesenchymal stem cells (MSCs) were differentiated into the adipogenic, osteogenic, and chondrogenic lineages. It was seen that perivascular cells, as well as CD146(-) cells and MSCs, cultured in stem cell medium showed greater differentiation than cells cultured in pericyte-specific medium. The expression of pericyte-specific markers CD146, neural/glial antigen 2 (NG2), platelet-derived growth factor receptor-β (PDGFR-β), myosin, and α-smooth muscle actin (α-SMA) could be found in both pericyte cultures, as well as to varying amounts in CD146(-) cells, MSCs, and endothelial cells. The here presented work shows that perivascular cells can adapt to their in vitro environment and cell culture conditions influence cell functionality, such as doubling rate or differentiation behavior. Pericyte-specific markers were shown to be expressed also from cells other than perivascular cells. We can further conclude that CD146(+) perivascular cells are inhomogeneous cell population probably containing stem cell subpopulations, which are located perivascular around capillaries. © 2015 International Federation for Cell Biology.

Biogelx: Cell Culture on Self-Assembling Peptide Gels.

Science.gov (United States)

Harper, Mhairi M; Connolly, Michael L; Goldie, Laura; Irvine, Eleanore J; Shaw, Joshua E; Jayawarna, Vineetha; Richardson, Stephen M; Dalby, Matthew J; Lightbody, David; Ulijn, Rein V

2018-01-01

Aromatic peptide amphiphiles can form self-supporting nanostructured hydrogels with tunable mechanical properties and chemical compositions. These hydrogels are increasingly applied in two-dimensional (2D) and three-dimensional (3D) cell culture, where there is a rapidly growing need to store, grow, proliferate, and manipulate naturally derived cells within a hydrated, 3D matrix. Biogelx Limited is a biomaterials company, created to commercialize these bio-inspired hydrogels to cell biologists for a range of cell culture applications. This chapter describes methods of various characterization and cell culture techniques specifically optimized for compatibility with Biogelx products.

Carrot Loss during Primary Production : Field Waste and Pack House Waste.

OpenAIRE

Bond, Rebekka

2016-01-01

Background: it has been suggested that roughly one-third of all food produced for human consumption is lost or wasted globally. The reduction of loss and waste is seen as an important societal issue with considerable ethical, ecological and economic implications. Fruit and vegetables have the highest wastage rates of any food products; (45 %). And a big part of this waste occurs during production, but empirical data on loss during primary production is limited. Carrots are an important hortic...

Development of a microfluidic perfusion 3D cell culture system

Science.gov (United States)

Park, D. H.; Jeon, H. J.; Kim, M. J.; Nguyen, X. D.; Morten, K.; Go, J. S.

2018-04-01

Recently, 3-dimensional in vitro cell cultures have gained much attention in biomedical sciences because of the closer relevance between in vitro cell cultures and in vivo environments. This paper presents a microfluidic perfusion 3D cell culture system with consistent control of long-term culture conditions to mimic an in vivo microenvironment. It consists of two sudden expansion reservoirs to trap incoming air bubbles, gradient generators to provide a linear concentration, and microchannel mixers. Specifically, the air bubbles disturb a flow in the microfluidic channel resulting in the instability of the perfusion cell culture conditions. For long-term stable operation, the sudden expansion reservoir is designed to trap air bubbles by using buoyancy before they enter the culture system. The performance of the developed microfluidic perfusion 3D cell culture system was examined experimentally and compared with analytical results. Finally, it was applied to test the cytotoxicity of cells infected with Ewing’s sarcoma. Cell death was observed for different concentrations of H2O2. For future work, the developed microfluidic perfusion 3D cell culture system can be used to examine the behavior of cells treated with various drugs and concentrations for high-throughput drug screening.

Cardiac Cells Beating in Culture: A Laboratory Exercise

Science.gov (United States)

Weaver, Debora

2007-01-01

This article describes how to establish a primary tissue culture, where cells are taken directly from an organ of a living animal. Cardiac cells are taken from chick embryos and transferred to culture dishes. These cells are not transformed and therefore have a limited life span. However, the unique characteristics of cardiac cells are maintained…

Improved endothelial cell seeding with cultured cells and fibronectin-coated grafts

International Nuclear Information System (INIS)

Seeger, J.M.; Klingman, N.

1985-01-01

A possible approach to the low seeding efficiency of endothelial cells into prosthetic grafts is to increase the number of cells to be seeded in cell culture and improve seeding efficiency by graft precoating with fibronectin. The effect of cell culture on cell adhesion is unknown, however, and fibronectin also binds fibrin, which may increase the thrombogenicity of the graft luminal surface. To investigate these questions, freshly harvested canine jugular vein endothelial cells from six animals and similar cells harvested from six primary and eight secondary cell cultures were labeled with 111 Indium and seeded into 5 cm, 4 mm PTFE grafts coated with fibronectin, using similar uncoated PTFE grafts as controls. Platelet accumulation and distribution on six similar coated and uncoated grafts placed in canine carotid, external jugular arterial venous shunts for 2 hr were also determined using autogenous 111 Indium-labeled platelets. Significant differences between group means were determined using the paired Student's t test. Results reveal that seeding efficiency is significantly better in all groups of coated grafts compared to uncoated grafts (P less than 0.01). Cells derived from cell culture also had significantly higher seeding efficiencies than freshly harvested cells when seeded into coated grafts (P less than 0.05) and tended to have higher seeding efficiencies than harvested cells when seeded into uncoated grafts (P = 0.53). Fibronectin coating increased mean platelet accumulation on the entire graft luminal surface, but not to a statistically significant degree (P greater than 0.1). Whether this increased seeding efficiency will improve graft endothelialization remains to be investigated

Dynamic cell culture system: a new cell cultivation instrument for biological experiments in space

Science.gov (United States)

Gmunder, F. K.; Nordau, C. G.; Tschopp, A.; Huber, B.; Cogoli, A.

1988-01-01

The prototype of a miniaturized cell cultivation instrument for animal cell culture experiments aboard Spacelab is presented (Dynamic cell culture system: DCCS). The cell chamber is completely filled and has a working volume of 200 microliters. Medium exchange is achieved with a self-powered osmotic pump (flowrate 1 microliter h-1). The reservoir volume of culture medium is 230 microliters. The system is neither mechanically stirred nor equipped with sensors. Hamster kidney (Hak) cells growing on Cytodex 3 microcarriers were used to test the biological performance of the DCCS. Growth characteristics in the DCCS, as judged by maximal cell density, glucose consumption, lactic acid secretion and pH, were similar to those in cell culture tubes.

Quantitative volumetric Raman imaging of three dimensional cell cultures

KAUST Repository

Kallepitis, Charalambos; Bergholt, Mads S.; Mazo, Manuel M.; Leonardo, Vincent; Skaalure, Stacey C.; Maynard, Stephanie A.; Stevens, Molly M.

2017-01-01

in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies

System-level modeling and simulation of the cell culture microfluidic biochip ProCell

DEFF Research Database (Denmark)

Minhass, Wajid Hassan; Pop, Paul; Madsen, Jan

2010-01-01

Microfluidic biochips offer a promising alternative to a conventional biochemical laboratory. There are two technologies for the microfluidic biochips: droplet-based and flow-based. In this paper we are interested in flow-based microfluidic biochips, where the liquid flows continuously through pre......-defined micro-channels using valves and pumps. We present an approach to the system-level modeling and simulation of a cell culture microfluidic biochip called ProCell, Programmable Cell Culture Chip. ProCell contains a cell culture chamber, which is envisioned to run 256 simultaneous experiments (viewed...

Culturing of PC12 Cells, Neuronal Cells, Astrocytes Cultures and Brain Slices in an Open Microfluidic System

DEFF Research Database (Denmark)

Al Atraktchi, Fatima Al-Zahraa; Bakmand, Tanya; Rømer Sørensen, Ane

The brain is the center of the nervous system, where serious neurodegenerative diseases such as Parkinson’s, Alzheimer’s and Huntington’s are products of functional loss in the neural cells (1). Typical techniques used to investigate these diseases lack precise control of the cellular surroundings......, in addition to isolating the neural tissue from nutrient delivery and to creating unwanted gradients (2). This means that typical techniques used to investigate neurodegenerative diseases cannot mimic in vivo conditions, as closely as desired. We have developed a novel microfluidic system for culturing PC12...... cells, neuronal cells, astrocytes cultures and brain slices. The microfluidic system provides efficient nutrient delivery, waste removal, access to oxygen, fine control over the neurochemical environment and access to modern microscopy. Additionally, the setup consists of an in vitro culturing...

Maintenance of mesenchymal stem cells culture due to the cells with reduced attachment rate

Directory of Open Access Journals (Sweden)

Shuvalova N. S.

2013-01-01

Full Text Available Aim. The classic detachment techniques lead to changes in cells properties. We offer a simple method of cultivating the population of cells that avoided an influence on the surface structures. Methods. Mesenchymal stem cells (MSC from human umbilical cord matrix were obtained and cultivated in standard conditions. While substituting the culture media by a fresh portion, the conditioned culture medium, where the cells were maintained for three days, was transferred to other culture flacks with addition of serum and growth factors. Results. In the flacks, one day after medium transfer, we observed attached cells with typical MSC morphology. The cultures originated from these cells had the same rate of surface markers expression and clonogenic potential as those replated by standard methods. Conclusions. MSC culture, derived by preserving the cells with reduced attachment ability, actually has the properties of «parent» passage. Using this method with accepted techniques of cells reseeding would allow maintaining the cells that avoided an impact on the cell surface proteins.

Evaluation of the osteogenic differentiation of gingiva-derived stem cells grown on culture plates or in stem cell spheroids: Comparison of two- and three-dimensional cultures.

Science.gov (United States)

Lee, Sung-Il; Ko, Youngkyung; Park, Jun-Beom

2017-09-01

Three-dimensional cell culture systems provide a convenient in vitro model for the study of complex cell-cell and cell-matrix interactions in the absence of exogenous substrates. The current study aimed to evaluate the osteogenic differentiation potential of gingiva-derived stem cells cultured in two-dimensional or three-dimensional systems. To the best of our knowledge, the present study is the first to compare the growth of gingiva-derived stem cells in monolayer culture to a three-dimensional culture system with microwells. For three-dimensional culture, gingiva-derived stem cells were isolated and seeded into polydimethylsiloxane-based concave micromolds. Alkaline phosphatase activity and alizarin red S staining assays were then performed to evaluate osteogenesis and the degree of mineralization, respectively. Stem cell spheroids had a significantly increased level of alkaline phosphatase activity and mineralization compared with cells from the two-dimensional culture. In addition, an increase in mineralized deposits was observed with an increase in the loading cell number. The results of present study indicate that gingiva-derived stem cell spheroids exhibit an increased osteogenic potential compared with stem cells from two-dimensional culture. This highlights the potential of three-dimensional culture systems using gingiva-derived stem cells for regenerative medicine applications requiring stem cells with osteogenic potential.

A novel three-dimensional cell culture method enhances antiviral drug screening in primary human cells.

Science.gov (United States)

Koban, Robert; Neumann, Markus; Daugs, Aila; Bloch, Oliver; Nitsche, Andreas; Langhammer, Stefan; Ellerbrok, Heinz

2018-02-01

Gefitinib is a specific inhibitor of the epidermal growth factor receptor (EGFR) and FDA approved for treatment of non-small cell lung cancer. In a previous study we could show the in vitro efficacy of gefitinib for treatment of poxvirus infections in monolayer (2D) cultivated cell lines. Permanent cell lines and 2D cultures, however, are known to be rather unphysiological; therefore it is difficult to predict whether determined effective concentrations or the drug efficacy per se are transferable to the in vivo situation. 3D cell cultures, which meanwhile are widely distributed across all fields of research, are a promising tool for more predictive in vitro investigations of antiviral compounds. In this study the spreading of cowpox virus and the antiviral efficacy of gefitinib were analyzed in primary human keratinocytes (NHEK) grown in a novel 3D extracellular matrix-based cell culture model and compared to the respective monolayer culture. 3D-cultivated NHEK grew in a polarized and thus a more physiological manner with altered morphology and close cell-cell contact. Infected cultures showed a strongly elevated sensitivity towards gefitinib. EGFR phosphorylation, cell proliferation, and virus replication were significantly reduced in 3D cultures at gefitinib concentrations which were at least 100-fold lower than those in monolayer cultures and well below the level of cytotoxicity. Our newly established 3D cell culture model with primary human cells is an easy-to-handle alternative to conventional monolayer cell cultures and previously described more complex 3D cell culture systems. It can easily be adapted to other cell types and a broad spectrum of viruses for antiviral drug screening and many other aspects of virus research under more in vivo-like conditions. In consequence, it may contribute to a more targeted realization of necessary in vivo experiments. Copyright © 2017 Elsevier B.V. All rights reserved.

Use of the software Seed Vigor Imaging System (SVIS® for assessing vigor of carrot seeds

Directory of Open Access Journals (Sweden)

José Luís de Marchi

Full Text Available ABSTRACT Seed vigor has traditionally been evaluated by physiological, biochemical and stress tolerance tests. More recently, with the use of computerized image analysis, objective information has become accessible in a relatively short period of time, with less human interference. The aim of this study was to verify the efficiency of computerized seedling image analysis by Seed Vigor Imaging System (SVIS® to detect differences in vigor between carrot (Daucus carota L. seed lots as compared to those provided by traditional vigor tests. Seeds from seven lots from the Brasilia cultivar were subjected to a germination test, first count of germination, speed of germination, accelerated aging with saline solution and seedling emergence; furthermore, a vigor index, growth index and uniformity index were determined by the Seed Vigor Imaging System (SVIS® during four evaluation periods. The results obtained by the computerized seedling analysis (vigor index and growth index show that SVIS® is efficient in assessing carrot seed vigor.

The Increased Content of Micronutrients in Celery, Carrot, Parsnip and Parsley Plants after Treatment with Sodium Naphthenate

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Grbović Ljubica

2016-08-01

Full Text Available Young plants of celery, parsley, parsnip and carrot, grown in nutrient solution, were treated with sodium naphthenate (10−7 mol dm−3, applying foliar and root treatments. Both treatments affected the root content of all investigated elements present in the nutrient solution, but in a different way, depending on the plant species. An average change (increase/decrease in the contents of investigated essential elements was about 35%. Our experiments with naphthenate showed that this treatment may enhance the efficiency of essential elements uptake and increase its content in plants without changing concentration of these elements in the nutrient solution. Especially interesting results were obtained in the case of carrot, as increased contents were observed in the elements that are usually deficient in nutrition (Fe, Zn, Mn, whereas the other remained unchanged.

Mesenchymal stem cells enhance the metastasis of 3D-cultured hepatocellular carcinoma cells

International Nuclear Information System (INIS)

Liu, Chang; Liu, Yang; Xu, Xiao-xi; Guo, Xin; Sun, Guang-wei; Ma, Xiao-jun

2016-01-01

Accumulating evidences have demonstrated that mesenchymal stem cells (MSC) could be recruited to the tumor microenvironment. Umbilical cord mesenchymal stem cells (UCMSC) were attractive vehicles for delivering therapeutic agents against cancer. Nevertheless, the safety of UCMSC in the treatment of tumors including hepatocellular carcinoma (HCC) was still undetermined. In this study, an in vitro co-culture system was established to evaluate the effect of UCMSC on the cell growth, cancer stem cell (CSC) characteristics, drug resistance, metastasis of 3D-cultured HCC cells, and the underlying mechanism was also investigated. It was found that after co-cultured with UCMSC, the metastatic ability of 3D-cultured HCC cells was significantly enhanced as indicated by up-regulation of matrix metalloproteinase (MMP), epithelial-mesenchymal transition (EMT)-related genes, and migration ability. However, cell growth, drug resistance and CSC-related gene expression of HCC cells were not affected by UCMSC. Moreover, EMT was reversed, MMP-2 expression was down-regulated, and migration ability of HCC cell was significantly inhibited when TGF-β receptor inhibitor SB431542 was added into the co-culture system. Therefore, these data indicated that UCMSC could significantly enhance the tumor cell metastasis, which was due to the EMT of HCC cells induced by TGF-β. The online version of this article (doi:10.1186/s12885-016-2595-4) contains supplementary material, which is available to authorized users

Primary Human Uterine Leiomyoma Cell Culture Quality Control: Some Properties of Myometrial Cells Cultured under Serum Deprivation Conditions in the Presence of Ovarian Steroids.

Science.gov (United States)

Bonazza, Camila; Andrade, Sheila Siqueira; Sumikawa, Joana Tomomi; Batista, Fabrício Pereira; Paredes-Gamero, Edgar J; Girão, Manoel J B C; Oliva, Maria Luiza V; Castro, Rodrigo Aquino

2016-01-01

Cell culture is considered the standard media used in research to emulate the in vivo cell environment. Crucial in vivo experiments cannot be conducted in humans and depend on in vitro methodologies such as cell culture systems. However, some procedures involving the quality control of cells in culture have been gradually neglected by failing to acknowledge that primary cells and cell lines change over time in culture. Thus, we report methods based on our experience for monitoring primary cell culture of human myometrial cells derived from uterine leiomyoma. We standardized the best procedure of tissue dissociation required for the study of multiple genetic marker systems that include species-specific antigens, expression of myofibroblast or myoblast markers, growth curve, serum deprivation, starvation by cell cycle synchronization, culture on collagen coated plates, and 17 β-estradiol (E2) and progesterone (P4) effects. The results showed that primary myometrial cells from patients with uterine leiomyoma displayed myoblast phenotypes before and after in vitro cultivation, and leiomyoma cells differentiated into mature myocyte cells under the appropriate differentiation-inducing conditions (serum deprivation). These cells grew well on collagen coated plates and responded to E2 and P4, which may drive myometrial and leiomyoma cells to proliferate and adhere into a focal adhesion complex involvement in a paracrine manner. The establishment of these techniques as routine procedures will improve the understanding of the myometrial physiology and pathogenesis of myometrium-derived diseases such as leiomyoma. Mimicking the in vivo environment of fibrotic conditions can prevent false results and enhance results that are based on cell culture integrity.

Relationship between moisture content and electrical impedance of carrot slices during drying

Science.gov (United States)

Kertész, Ákos; Hlaváčová, Zuzana; Vozáry, Eszter; Staroňová, Lenka

2015-01-01

Electrical properties of food materials can give information about the inner structure and physiological state of biological tissues. Generally, the process of drying of fruits and vegetables is followed by weight loss. The aim of this study was to measure the impedance spectra of carrot slices during drying and to correlate impedance parameters to moisture content in different drying periods. Cylindrical slices were cut out from the carrot root along the axis. The slices were dried in a Venticell 111 air oven at 50°C. The weight of the slices was measured with a Denver SI-603 electronic analytical and precision balance. The weighing of the samples was performed every 30 min at the beginning of drying and every 60 min after the process. The moisture content of the samples was calculated on wet basis. The magnitude and phase angle of electrical impedance of the slices were measured with HP 4284A and 4285A precision LCR meters in the frequency range from 30 Hz to 1 MHz and from 75 kHz to 30 MHz, respectively, at voltage 1 V. The impedance measurement was performed after weighting. The change in the magnitude of impedance during drying showed a good correlation with the change in the moisture content.

Microbiological nutritional and sensorial changes in fresh carrot juice preserved by irradiation

International Nuclear Information System (INIS)

Hammad, A. A. I.; Abd-El-kalek, H.; Abd-El-kader, R. M.; Youssef, K. H.

2012-12-01

Fresh carroll juice have perishable nature and very limited shelf-life, and may poses a microbiological hazard. Gamma irradiation (1.5, 3.0 and 4.0 kGy ) as non-thermal proceeding was performed to improve microbial quality, ensure safety and extending the refrigerated shelf-life of fresh carrot juice. Although irradiation dose of 1.5 kGy had no significant effect on total carotenoids, physicochemical and sensory properties of the juice, it significantly reduced ascorbic acid content and was not enough for reducing microbial content to great extend and for eliminating enterococcus fecal it.However, there was no significant difference in total carotene content between all irradiated samples and non-irradiated control. It was found that immediately after irradiation the sensory scores of irradiated (1.5 and 3.0 kGy) and non-irradiation samples were not significantly different. Irradiation dose of 4.0 kGy significantly reduced the nutrition content and the sensory quality attributes of the juice. Thus, irradiation dose of 3.0 kGy can be successfully used to improve the microbial quality and extend the refrigerated shelf-life of fresh carrot juice, where it extended the shelf-life to 8 days against only 2 day for non-irradiated control. (Author)

Metabolomics for organic food authentication: Results from a long-term field study in carrots.

Science.gov (United States)

Cubero-Leon, Elena; De Rudder, Olivier; Maquet, Alain

2018-01-15

Increasing demand for organic products and their premium prices make them an attractive target for fraudulent malpractices. In this study, a large-scale comparative metabolomics approach was applied to investigate the effect of the agronomic production system on the metabolite composition of carrots and to build statistical models for prediction purposes. Orthogonal projections to latent structures-discriminant analysis (OPLS-DA) was applied successfully to predict the origin of the agricultural system of the harvested carrots on the basis of features determined by liquid chromatography-mass spectrometry. When the training set used to build the OPLS-DA models contained samples representative of each harvest year, the models were able to classify unknown samples correctly (100% correct classification). If a harvest year was left out of the training sets and used for predictions, the correct classification rates achieved ranged from 76% to 100%. The results therefore highlight the potential of metabolomic fingerprinting for organic food authentication purposes. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Construction of 2D transparent micromodels in polyester resin with porosity similar to carrots

Directory of Open Access Journals (Sweden)

Rodrigo Emilio Díaz

2011-12-01

Full Text Available Microscopic visualization, especially in transparent micromodels, can provide valuable information to understand the transport phenomena at pore scale in different process occurring in porous materials (food, timber, soils, etc.. Micromodels studies focus mainly on the observation of multi-phase flow, which presents a greater proximity to reality. The aim of this study was to study the process of flexography and its application in the manufacture of polyester resin transparent micromodels and its application to carrots. Materials used to implement a flexo station for micromodels construction were thermoregulated water bath, exposure chamber to UV light, photosensitive substance (photopolymer, RTV silicone polyester resin, and glass plates. In this paper, data on size distribution of a particular kind of carrot we used, and a transparent micromodel with square cross-section as well as a Log-normal pore size distribution with pore radii ranging from 10 to 110 µm (average of 22 µm and micromodel size of 10 × 10 cm were built. Finally, it stresses that it has successfully implemented the protocol processing 2D polyester resin transparent micromodels.

Radiosensitivity of normal human epidermal cells in culture

International Nuclear Information System (INIS)

Dover, R.; Potten, C.S.

1983-01-01

Using an in vitro culture system the authors have derived #betta#-radiation survival curves over a dose range 0-8 Gy for the clonogenic cells of normal human epidermis. The culture system used allows the epidermal cells to stratify and form a multi-layered sheet of keratinizing cells. The cultures appear to be a very good model for epidermis in vivo. The survival curves show a population which is apparently more sensitive than murine epidermis in vivo. It remains unclear whether this is an intrinsic difference between the species or is a consequence of the in vitro cultivation of the human cells. (author)

Differential selection on carotenoid biosynthesis genes as a function of gene position in the metabolic pathway: a study on the carrot and dicots.

Directory of Open Access Journals (Sweden)

Jérémy Clotault

Full Text Available Selection of genes involved in metabolic pathways could target them differently depending on the position of genes in the pathway and on their role in controlling metabolic fluxes. This hypothesis was tested in the carotenoid biosynthesis pathway using population genetics and phylogenetics.Evolutionary rates of seven genes distributed along the carotenoid biosynthesis pathway, IPI, PDS, CRTISO, LCYB, LCYE, CHXE and ZEP, were compared in seven dicot taxa. A survey of deviations from neutrality expectations at these genes was also undertaken in cultivated carrot (Daucus carota subsp. sativus, a species that has been intensely bred for carotenoid pattern diversification in its root during its cultivation history. Parts of sequences of these genes were obtained from 46 individuals representing a wide diversity of cultivated carrots. Downstream genes exhibited higher deviations from neutral expectations than upstream genes. Comparisons of synonymous and nonsynonymous substitution rates between genes among dicots revealed greater constraints on upstream genes than on downstream genes. An excess of intermediate frequency polymorphisms, high nucleotide diversity and/or high differentiation of CRTISO, LCYB1 and LCYE in cultivated carrot suggest that balancing selection may have targeted genes acting centrally in the pathway.Our results are consistent with relaxed constraints on downstream genes and selection targeting the central enzymes of the carotenoid biosynthesis pathway during carrot breeding history.

Differentiation of oligodendrocyte progenitor cells from dissociated monolayer and feeder-free cultured pluripotent stem cells.

Science.gov (United States)

Yamashita, Tomoko; Miyamoto, Yuki; Bando, Yoshio; Ono, Takashi; Kobayashi, Sakurako; Doi, Ayano; Araki, Toshihiro; Kato, Yosuke; Shirakawa, Takayuki; Suzuki, Yutaka; Yamauchi, Junji; Yoshida, Shigetaka; Sato, Naoya

2017-01-01

Oligodendrocytes myelinate axons and form myelin sheaths in the central nervous system. The development of therapies for demyelinating diseases, including multiple sclerosis and leukodystrophies, is a challenge because the pathogenic mechanisms of disease remain poorly understood. Primate pluripotent stem cell-derived oligodendrocytes are expected to help elucidate the molecular pathogenesis of these diseases. Oligodendrocytes have been successfully differentiated from human pluripotent stem cells. However, it is challenging to prepare large amounts of oligodendrocytes over a short amount of time because of manipulation difficulties under conventional primate pluripotent stem cell culture methods. We developed a proprietary dissociated monolayer and feeder-free culture system to handle pluripotent stem cell cultures. Because the dissociated monolayer and feeder-free culture system improves the quality and growth of primate pluripotent stem cells, these cells could potentially be differentiated into any desired functional cells and consistently cultured in large-scale conditions. In the current study, oligodendrocyte progenitor cells and mature oligodendrocytes were generated within three months from monkey embryonic stem cells. The embryonic stem cell-derived oligodendrocytes exhibited in vitro myelinogenic potency with rat dorsal root ganglion neurons. Additionally, the transplanted oligodendrocyte progenitor cells differentiated into myelin basic protein-positive mature oligodendrocytes in the mouse corpus callosum. This preparative method was used for human induced pluripotent stem cells, which were also successfully differentiated into oligodendrocyte progenitor cells and mature oligodendrocytes that were capable of myelinating rat dorsal root ganglion neurons. Moreover, it was possible to freeze, thaw, and successfully re-culture the differentiating cells. These results showed that embryonic stem cells and human induced pluripotent stem cells maintained in a

Culture and Characterization of Circulating Endothelial Progenitor Cells in Patients with Renal Cell Carcinoma.

Science.gov (United States)

Gu, Wenyu; Sun, Wei; Guo, Changcheng; Yan, Yang; Liu, Min; Yao, Xudong; Yang, Bin; Zheng, Junhua

2015-07-01

Although emerging evidence demonstrates increased circulating endothelial progenitor cells in patients with solid tumors, to our knowledge it is still unknown whether such cells can be cultured from patients with highly angiogenic renal cell carcinoma. We cultured and characterized circulating endothelial progenitor cells from patients with renal cell carcinoma. The circulating endothelial progenitor cell level (percent of CD45(-)CD34(+) VEGF-R2(+) cells in total peripheral blood mononuclear cells) was quantified in 47 patients with renal cell carcinoma and 40 healthy controls. Peripheral blood mononuclear cells were then isolated from 33 patients with renal cell carcinoma and 30 healthy controls to culture and characterize circulating endothelial progenitor cells. The circulating endothelial progenitor cell level was significantly higher in patients with renal cell carcinoma than in healthy controls (0.276% vs 0.086%, p cells first emerged significantly earlier in patient than in control preparations (6.72 vs 14.67 days, p culture success rate (87.8% vs 40.0% of participants) and the number of colonies (10.06 vs 1.83) were significantly greater for patients than for controls (each p cell level correlated positively with the number of patient colonies (r = 0.762, p Cells cultured from patients and controls showed a similar growth pattern, immunophenotype, ability to uptake Ac-LDL and bind lectin, and form capillary tubes in vitro. However, significantly more VEGF-R2(+) circulating endothelial progenitor cells were found in preparations from patients with renal cell carcinoma than from healthy controls (21.1% vs 13.4%, p cell colonies, a higher cell culture success rate and more colonies were found for patients with renal cell carcinoma than for healthy controls. Results indicate the important significance of VEGF-R2(+) circulating endothelial progenitors in patients with renal cell carcinoma. Copyright © 2015 American Urological Association Education and Research

The impact of cell culture equipment on energy loss.

Science.gov (United States)

Davies, Lleucu B; Kiernan, Michael N; Bishop, Joanna C; Thornton, Catherine A; Morgan, Gareth

2014-01-01

Light energy of discrete wavelengths supplied via lasers and broadband intense pulsed light have been used therapeutically for many years. In vitro models complement clinical studies, especially for the elucidation of underlying mechanisms of action. Clarification that light energy reaches the cells is necessary when developing protocols for the treatment of cells using in vitro models. Few studies report on energy loss in cell culture equipment. The ability of energy from light with therapeutic potential to reach cells in culture needs to be determined; this includes determining the proportion of light energy lost within standard cell culture media and cell culture vessels. The energy absorption of cell culture media, with/without the pH indicator dye phenol red, and the loss of energy within different plastics and glassware used typically for in vitro cell culture were investigated using intense pulsed light and a yellow pulsed dye laser. Media containing phenol red have a distinctive absorption peak (560 nm) absent in phenol red-free media and restored by the addition of phenol red. For both light sources, energy loss was lowest in standard polystyrene tissue culture flasks or multi-well plates and highest in polypropylene vessels or glass tubes. The effects of phenol red-free media on the absorption of energy varied with the light source used. Phenol red-free media are the media of choice; polystyrene vessels with flat surfaces such as culture flasks or multi-well plates should be used in preference to polypropylene or glass vessels.

Surface modified alginate microcapsules for 3D cell culture

Science.gov (United States)

Chen, Yi-Wen; Kuo, Chiung Wen; Chueh, Di-Yen; Chen, Peilin

2016-06-01

Culture as three dimensional cell aggregates or spheroids can offer an ideal platform for tissue engineering applications and for pharmaceutical screening. Such 3D culture models, however, may suffer from the problems such as immune response and ineffective and cumbersome culture. This paper describes a simple method for producing microcapsules with alginate cores and a thin shell of poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) to encapsulate mouse induced pluripotent stem (miPS) cells, generating a non-fouling surface as an effective immunoisolation barrier. We demonstrated the trapping of the alginate microcapsules in a microwell array for the continuous observation and culture of a large number of encapsulated miPS cells in parallel. miPS cells cultured in the microcapsules survived well and proliferated to form a single cell aggregate. Droplet formation of monodisperse microcapsules with controlled size combined with flow cytometry provided an efficient way to quantitatively analyze the growth of encapsulated cells in a high-throughput manner. The simple and cost-effective coating technique employed to produce the core-shell microcapsules could be used in the emerging field of cell therapy. The microwell array would provide a convenient, user friendly and high-throughput platform for long-term cell culture and monitoring.

A Simple Hydrophilic Treatment of SU-8 Surfaces for Cell Culturing and Cell Patterning

DEFF Research Database (Denmark)

Wang, Zhenyu; Stangegaard, Michael; Dufva, Hans Martin

2005-01-01

SU-8, an epoxy-based photoresist, widely used in constitution different mTAS systems, is incompatible with mammalian cell adhesion and culture in its native form. Here, we demonstrate a simple, cheap and robust two-step method to render a SU-8 surface hydrophilic and compatible with cell culture........ The contact angle of SU-8 surface was significantly reduced from 90° to 25° after the surface modification. The treated SU-8 surfaces provided a cell culture environment that was comparable with cell culture flask surface in terms of generation time and morphology....

Herbicide contamination in carrot grown in punjab, pakistan

International Nuclear Information System (INIS)

Amjad, M.; Ahmad, T.; Jahangir, M.M.

2013-01-01

Food safety and security is a burning issue of the time whereas vegetable production is an important aspect of agriculture. Use of herbicides for vegetable production is very common in Pakistan but no proper procedure has been planned to keep optimal level of doses of herbicide under permissible limit. To estimate the pesticide residues, samples from the leading carrot producing sites were collected along with the samples from the market. The samples were processed using standard procedures and qualitative and quantitative analysis was performed by Gas Chromatography-Mass Spectrometry (GC-MS). It was concluded that all the samples were contaminated with S-metolachlor in the range of 0.45 to 0.73 mg kg-1 which was above the permissible limit (0.40 mg kg-1). (author)

Identification of differences in gene expression in primary cell cultures of human endometrial epithelial cells and trophoblast cells following their interaction

DEFF Research Database (Denmark)

Høgh, Mette; Islin, Henrik; Møller, Charlotte

2006-01-01

The interaction between the cell types was simulated in vitro by growing primary cell cultures of human endometrial epithelial cells and trophoblast cells together (co-culture) and separately (control cultures). Gene expression in the cell cultures was compared using the Differential Display method and confirmed...

Establishment of automated culture system for murine induced pluripotent stem cells

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Koike Hiroyuki

2012-11-01

Full Text Available Abstract Background Induced pluripotent stem (iPS cells can differentiate into any cell type, which makes them an attractive resource in fields such as regenerative medicine, drug screening, or in vitro toxicology. The most important prerequisite for these industrial applications is stable supply and uniform quality of iPS cells. Variation in quality largely results from differences in handling skills between operators in laboratories. To minimize these differences, establishment of an automated iPS cell culture system is necessary. Results We developed a standardized mouse iPS cell maintenance culture, using an automated cell culture system housed in a CO2 incubator commonly used in many laboratories. The iPS cells propagated in a chamber uniquely designed for automated culture and showed specific colony morphology, as for manual culture. A cell detachment device in the system passaged iPS cells automatically by dispersing colonies to single cells. In addition, iPS cells were passaged without any change in colony morphology or expression of undifferentiated stem cell markers during the 4 weeks of automated culture. Conclusions Our results show that use of this compact, automated cell culture system facilitates stable iPS cell culture without obvious effects on iPS cell pluripotency or colony-forming ability. The feasibility of iPS cell culture automation may greatly facilitate the use of this versatile cell source for a variety of biomedical applications.

Genetic structure in cultivated and wild carrots (¤Daucus carota¤ L.) revealed by AFLP analysis

DEFF Research Database (Denmark)

Shim, S.I.; Bagger Jørgensen, Rikke

2000-01-01

Genetic variation within and among five Danish populations of wild carrot and five cultivated varieties was investigated using amplified fragment length polymorphism (AFLP). Ten AFLP primer combinations produced 116 polymorphic bands. Based on the marker data an UPGMA-cluster analysis and principal...

Youth Culture and Cell Phone

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mohammad saeed zokaei

2009-11-01

Full Text Available Iranian youth’s leisure culture has been immediately affected by the digital media culture. As a communicative media, cell phone has crossed borders of youth norms and identity; and in addition to facilitating their communication, has changed its patterns. Applying Bourdieu’s concepts of habitus and field, and relied on the qualitative and quantitative data gathered from the mobile youth users, the present study argues that mobile has produced a new field in which youth’s opportunities for leisure, entertainment, communication, and independence have extended. In addition, cell phone has facilitated and compensated for some defects in public sphere, and therefore empowered youth agency, individuality, and power. Despite this strengthening, cell phone does not cross borders of gender and class differences, or the levels of social capital.

Primary Human Uterine Leiomyoma Cell Culture Quality Control: Some Properties of Myometrial Cells Cultured under Serum Deprivation Conditions in the Presence of Ovarian Steroids.

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Camila Bonazza

Full Text Available Cell culture is considered the standard media used in research to emulate the in vivo cell environment. Crucial in vivo experiments cannot be conducted in humans and depend on in vitro methodologies such as cell culture systems. However, some procedures involving the quality control of cells in culture have been gradually neglected by failing to acknowledge that primary cells and cell lines change over time in culture. Thus, we report methods based on our experience for monitoring primary cell culture of human myometrial cells derived from uterine leiomyoma. We standardized the best procedure of tissue dissociation required for the study of multiple genetic marker systems that include species-specific antigens, expression of myofibroblast or myoblast markers, growth curve, serum deprivation, starvation by cell cycle synchronization, culture on collagen coated plates, and 17 β-estradiol (E2 and progesterone (P4 effects. The results showed that primary myometrial cells from patients with uterine leiomyoma displayed myoblast phenotypes before and after in vitro cultivation, and leiomyoma cells differentiated into mature myocyte cells under the appropriate differentiation-inducing conditions (serum deprivation. These cells grew well on collagen coated plates and responded to E2 and P4, which may drive myometrial and leiomyoma cells to proliferate and adhere into a focal adhesion complex involvement in a paracrine manner. The establishment of these techniques as routine procedures will improve the understanding of the myometrial physiology and pathogenesis of myometrium-derived diseases such as leiomyoma. Mimicking the in vivo environment of fibrotic conditions can prevent false results and enhance results that are based on cell culture integrity.

Bioaccessibility and arsenic speciation in carrots, beets and quinoa from a contaminated area of Chile

Energy Technology Data Exchange (ETDEWEB)

Pizarro, Isabel [Facultad de Ciencias Básicas, Universidad de Antofagasta, 02800 Antofagasta (Chile); Gómez-Gómez, Milagros [Department of Analytical Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, 28040 Madrid (Spain); León, Jennifer; Román, Domingo [Facultad de Ciencias Básicas, Universidad de Antofagasta, 02800 Antofagasta (Chile); Palacios, M. Antonia, E-mail: palacor@ucm.es [Department of Analytical Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, 28040 Madrid (Spain)

2016-09-15

Consumption of vegetables grown in arsenic (As)-contaminated soils is an important exposure route to the element for humans. The present study is focused on locally-grown, frequently-consumed vegetables, such as carrots (Daucus carota), beets (Beta vulgaris) and quinoa (Chenopodium) from the As-polluted Chiu Chiu area in Northern Chile. The latter region is affected both by As discharge from copper mining activity and natural As contamination, leading to a high As content in local food and water. For the selected vegetables, the following aspects were investigated: i) Their total As, Cu, Pb, Cr, Cd and Mn content; ii) Arsenic speciation in the edible part of the vegetables by liquid chromatography inductively-coupled plasma mass spectrometry (LC-ICPMS) analysis; iii) Arsenic bioaccessibility in the vegetables during in vitro gastrointestinal digestion; iv) Arsenic species present in the extracts obtained from in vitro gastrointestinal digestion; and v) Arsenic dietary exposure estimates for the assessment of the risk posed by the vegetables consumption. A significant degree of As contamination was found in the vegetables under study, their metal content having been compared with that of similar Spanish uncontaminated products. In vitro gastrointestinal digestion of the studied vegetables led to quantitative extraction of As from carrots and beets, whereas efficiency was about 40% for quinoa. For carrots, only As(III) and As(V) species were found, being their concentration levels similar. In the case of quinoa, around 85% of the element was present as As(V). For beets, inorganic As(V) and unknown overlapped As species (probably arsenosugars) were found. No significant transformation of the original As species was observed during in vitro gastrointestinal digestion. Arsenic dietary exposure values obtained for the three vegetables (0.017–0.021 μg As person{sup −1} day{sup −1}) were much lower than the JFCFA's safety limit of 50 μg As person{sup −1} day

Bioaccessibility and arsenic speciation in carrots, beets and quinoa from a contaminated area of Chile

International Nuclear Information System (INIS)

Pizarro, Isabel; Gómez-Gómez, Milagros; León, Jennifer; Román, Domingo; Palacios, M. Antonia

2016-01-01

Consumption of vegetables grown in arsenic (As)-contaminated soils is an important exposure route to the element for humans. The present study is focused on locally-grown, frequently-consumed vegetables, such as carrots (Daucus carota), beets (Beta vulgaris) and quinoa (Chenopodium) from the As-polluted Chiu Chiu area in Northern Chile. The latter region is affected both by As discharge from copper mining activity and natural As contamination, leading to a high As content in local food and water. For the selected vegetables, the following aspects were investigated: i) Their total As, Cu, Pb, Cr, Cd and Mn content; ii) Arsenic speciation in the edible part of the vegetables by liquid chromatography inductively-coupled plasma mass spectrometry (LC-ICPMS) analysis; iii) Arsenic bioaccessibility in the vegetables during in vitro gastrointestinal digestion; iv) Arsenic species present in the extracts obtained from in vitro gastrointestinal digestion; and v) Arsenic dietary exposure estimates for the assessment of the risk posed by the vegetables consumption. A significant degree of As contamination was found in the vegetables under study, their metal content having been compared with that of similar Spanish uncontaminated products. In vitro gastrointestinal digestion of the studied vegetables led to quantitative extraction of As from carrots and beets, whereas efficiency was about 40% for quinoa. For carrots, only As(III) and As(V) species were found, being their concentration levels similar. In the case of quinoa, around 85% of the element was present as As(V). For beets, inorganic As(V) and unknown overlapped As species (probably arsenosugars) were found. No significant transformation of the original As species was observed during in vitro gastrointestinal digestion. Arsenic dietary exposure values obtained for the three vegetables (0.017–0.021 μg As person"−"1 day"−"1) were much lower than the JFCFA's safety limit of 50 μg As person"−"1 day"−"1. Therefore

Cell culture density affects the proliferation activity of human adipose tissue stem cells.

Science.gov (United States)

Kim, Dae Seong; Lee, Myoung Woo; Ko, Young Jong; Chun, Yong Hoon; Kim, Hyung Joon; Sung, Ki Woong; Koo, Hong Hoe; Yoo, Keon Hee

2016-01-01

In this study, we investigated the effect of cell density on the proliferation activity of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT-MSCs) over time in culture. Passage #4 (P4) and #12 (P12) AT-MSCs from two donors were plated at a density of 200 (culture condition 1, CC1) or 5000 (culture condition 2, CC2) cells cm(-2) . After 7 days of incubation, P4 and P12 AT-MSCs cultured in CC1 were thin and spindle-shaped, whereas those cultured in CC2 had extensive cell-to-cell contacts and an expanded cell volume. In addition, P4 and P12 AT-MSCs in CC1 divided more than three times, while those in CC2 divided less than once on average. Flow cytometric analysis using 5(6)-carboxyfluorescein diacetate N-succinimidyl ester dye showed that the fluorescence intensity of AT-MSCs was lower in CC1 than in CC2. Furthermore, expression of proliferation-associated genes, such as CDC45L, CDC20A and KIF20A, in P4 AT-MSCs was higher in CC1 than in CC2, and this difference was also observed in P12 AT-MSCs. These data demonstrated that cell culture density affects the proliferation activity of MSCs, suggesting that it is feasible to design a strategy to prepare suitable MSCs using specific culture conditions. Copyright © 2016 John Wiley & Sons, Ltd.

Nanotechnology, Cell Culture and Tissue Engineering

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Kazutoshi Haraguchi

2011-01-01

Full Text Available We have fabricated new types of polymer hydrogels and polymer nanocomposites, i.e., nanocomposite gels (NC gels and soft, polymer nanocomposites (M-NCs: solid, with novel organic/inorganic network structures. Both NC gels and M-NCs were synthesized by in-situ free-radical polymerization in the presence of exfoliated clay platelets in aqueous systems and were obtained in various forms such as film, sheet, tube, coating, etc. and sizes with a wide range of clay contents. Here, disk-like inorganic clay nanoparticles act as multi-functional crosslinkers to form new types of network systems. Both NC gels and M-NCs have extraordinary optical and mechanical properties including ultra-high reversible extensibility, as well as a number of new characteristics relating to optical anisotropy, polymer/clay morphology, biocompatibility, stimuli-sensitive surfaces, micro-patterning, etc. For examples, the biological testing of medical devices, comprised of a sensitization test, an irritation test, an intracutaneous test and an in vitro cytotoxicity test,was carried out for NC gels and M-NCs. The safety of NC gels and M-NCs was confirmed in all tests. Also, the interaction of living tissue with NC gel was investigated in vivo by implantation in live goats; neither inflammation nor concrescence occurred around the NC gels. Furthermore, it was found that both N-NC gels consisting of poly(N-isopropylacrylamide(PNIPA/clay network and M-NCs consisting of poly(2-methoxyethyacrylate(PMEA/clay network show characteristic cell culture and subsequent cell detachment on their surfaces, although it was almost impossible to culture cells on conventional, chemically-crosslinked PNIPA hydrogels and chemically crossslinked PMEA, regardless of their crosslinker concentration. Various kinds of cells, such ashumanhepatoma cells (HepG2, normal human dermal fibroblast (NHDF, and human umbilical vein endothelial cells (HUVEC, could be cultured to be confluent on the surfaces of N

Qualidade de raízes de cenoura em sistemas consorciados com alface sob diferentes densidades populacionais Quality of carrot roots in intercropped systems with lettuce under different planting densities

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Aurélio P. Barros Júnior

2005-06-01

Full Text Available O experimento foi realizado de junho a setembro de 2003, em campo da ESAM, com o objetivo de avaliar a qualidade de raízes de cenoura (Brasília em sistemas consorciados com alface (Tainá sob diferentes densidades populacionais. O delineamento experimental utilizado foi de blocos casualizados, em esquema fatorial 4 x 4, com três repetições. Os tratamentos foram resultantes da combinação de quatro populações de cenoura [(40%, 60%, 80% e 100% da população recomendada no cultivo solteiro (PRCS] com quatro populações de alface (40%, 60%, 80% e 100% da PRCS. As características avaliadas nas raízes da cenoura foram acidez total titulável (ATT, sólidos solúveis totais (SST, açúcares totais (ACT pH, relação sólidos solúveis totais e acidez total titulável, além da produtividade comercial da cenoura e da alface. Ocorreu interação significativa entre as densidades populacionais das culturas componentes na ATT das raízes da cenoura. Houve aumento no conteúdo de SST e no pH à medida que se aumentou a densidade populacional de cenoura. Houve também um aumento na produtividade comercial de raízes de cenoura e na produtividade da alface com o aumento de suas densidades populacionais, respectivamente. A variação na população de cenoura não afetou a produtividade da alface, mas a variação na população da alface afetou negativamente a produtividade comercial da cenoura. Entre as características avaliadas, as que se correlacionaram de maneira significativa com a produtividade comercial de cenoura, foram as concentrações de sólidos solúveis totais e açucares totais, evidenciando assim, que estes atributos podem ser considerados representativos da qualidade de raízes da cenoura.The experiment was carried out from June to September 2003, in Mossoró, Rio Grande do Norte State, Brazil, to evaluate the quality of carrot roots (cv. Brasília in intercropped systems with lettuce (cv. Tainá under different planting

Recombinant Protein Production and Insect Cell Culture and Process

Science.gov (United States)

Spaulding, Glenn F. (Inventor); Goodwin, Thomas J. (Inventor); OConnor, Kim C. (Inventor); Francis, Karen M. (Inventor); Andrews, Angela D. (Inventor); Prewett, Tracey L. (Inventor)

1997-01-01

A process has been developed for recombinant production of selected polypeptides using transformed insect cells cultured in a horizontally rotating culture vessel modulated to create low shear conditions. A metabolically transformed insect cell line is produced using the culture procedure regardless of genetic transformation. The recombinant polypeptide can be produced by an alternative process using virtually infected or stably transformed insect cells containing a gene encoding the described polypeptide. The insect cells can also be a host for viral production.

Metabolite profiling of microfluidic cell culture conditions for droplet based screening

DEFF Research Database (Denmark)

Björk, Sara M.; Sjoström, Staffan L.; Svahn, Helene Andersson

2015-01-01

We investigate the impact of droplet culture conditions on cell metabolic state by determining key metabolite concentrations in S. cerevisiae cultures in different microfluidic droplet culture formats. Control of culture conditions is critical for single cell/clone screening in droplets......, such as directed evolution of yeast, as cell metabolic state directly affects production yields from cell factories. Here, we analyze glucose, pyruvate, ethanol, and glycerol, central metabolites in yeast glucose dissimilation to establish culture formats for screening of respiring as well as fermenting yeast...... limited cultures, whereas the metabolite profiles of cells cultured in the alternative wide tube droplet incubation format resemble those from aerobic culture. Furthermore, we demonstrate retained droplet stability and size in the new better oxygenated droplet incubation format....

A Cell Culture Approach to Optimized Human Corneal Endothelial Cell Function

Science.gov (United States)

Bartakova, Alena; Kuzmenko, Olga; Alvarez-Delfin, Karen; Kunzevitzky, Noelia J.; Goldberg, Jeffrey L.

2018-01-01

Purpose Cell-based therapies to replace corneal endothelium depend on culture methods to optimize human corneal endothelial cell (HCEC) function and minimize endothelial-mesenchymal transition (EnMT). Here we explore contribution of low-mitogenic media on stabilization of phenotypes in vitro that mimic those of HCECs in vivo. Methods HCECs were isolated from cadaveric donor corneas and expanded in vitro, comparing continuous presence of exogenous growth factors (“proliferative media”) to media without those factors (“stabilizing media”). Identity based on canonical morphology and expression of surface marker CD56, and function based on formation of tight junction barriers measured by trans-endothelial electrical resistance assays (TEER) were assessed. Results Primary HCECs cultured in proliferative media underwent EnMT after three to four passages, becoming increasingly fibroblastic. Stabilizing the cells before each passage by switching them to a media low in mitogenic growth factors and serum preserved canonical morphology and yielded a higher number of cells. HCECs cultured in stabilizing media increased both expression of the identity marker CD56 and also tight junction monolayer integrity compared to cells cultured without stabilization. Conclusions HCECs isolated from donor corneas and expanded in vitro with a low-mitogenic media stabilizing step before each passage demonstrate more canonical structural and functional features and defer EnMT, increasing the number of passages and total canonical cell yield. This approach may facilitate development of HCEC-based cell therapies. PMID:29625488

Biochemical studies on weaning foods based legumes and carrots

International Nuclear Information System (INIS)

Nabag, Fatima Omer

1997-01-01

Attempts were made to utilize available food sources in formulation of weaning foods. Common legumes (chick pea and pigeon pea ) were used as protein source. While dehydrated carrots powder were used as a vitamin A source. Addition of 25% chick pea increased the protein content of the weaning foods to 16.7% and 14.4%,respectively ,while Cerelac and Riri gave protein content of 15.3% and 7.3%,respectively. In corporation of carrots at 10% level gave a vitamin A content of 564 RE/100 g material. Weaning food containing chick pea recorded higher preference among panelists and significantly better (p≤0.05)than samples containing pigeon pea. The bulk density of newly developed based formulae CP 3 , PP 3 (0.7 g/ml, 0.8 g/ml, respectively ) was higher than the market weaning food Cerelac and Riri (0.6 g/ml and 0.5 g/ml,respectively )The formula CP 3 recorded lower hot paste viscosity (3500 cp.) than both values obtained for Cerelac (4500 cp.) and Riri (extremely viscous). The lysine content of CP 3 (3.9 g/100 g protein) was higher than respective values in market foods and for better when calculated per weaning material (0.65 g/100 g material) compared to the other products (0.57; 0.28 g/100 material of Cerelac and Riri, respectively). Chick pea-based formula (CP 3 ) was also found to possess higher in vitro protein didestibility (95.2%) compared to Cerelac (94.2%) and Riri (88.5%). The calculated protein efficiency ratio (C-PER) of CP 3 (1.7) was higher than that of Riri (1.6) and lower than that of cerelac (2.7). (Author)

Biochemical studies on weaning foods based legumes and carrots

Energy Technology Data Exchange (ETDEWEB)

Nabag, Fatima Omer [Department of Food Science and Technology, Faculty of Agriculture, University of Khartoum, Khartoum (Sudan)

1998-12-31

Attempts were made to utilize available food sources in formulation of weaning foods. Common legumes (chick pea and pigeon pea ) were used as protein source. While dehydrated carrots powder were used as a vitamin A source. Addition of 25% chick pea increased the protein content of the weaning foods to 16.7% and 14.4%,respectively ,while Cerelac and Riri gave protein content of 15.3% and 7.3%,respectively. In corporation of carrots at 10% level gave a vitamin A content of 564 RE/100 g material. Weaning food containing chick pea recorded higher preference among panelists and significantly better (p{<=}0.05)than samples containing pigeon pea. The bulk density of newly developed based formulae CP{sup 3}, PP{sup 3} (0.7 g/ml, 0.8 g/ml, respectively ) was higher than the market weaning food Cerelac and Riri (0.6 g/ml and 0.5 g/ml,respectively )The formula CP{sup 3} recorded lower hot paste viscosity (3500 cp.) than both values obtained for Cerelac (4500 cp.) and Riri (extremely viscous). The lysine content of CP{sup 3} (3.9 g/100 g protein) was higher than respective values in market foods and for better when calculated per weaning material (0.65 g/100 g material) compared to the other products (0.57; 0.28 g/100 material of Cerelac and Riri, respectively). Chick pea-based formula (CP{sup 3}) was also found to possess higher in vitro protein didestibility (95.2%) compared to Cerelac (94.2%) and Riri (88.5%). The calculated protein efficiency ratio (C-PER) of CP{sup 3} (1.7) was higher than that of Riri (1.6) and lower than that of cerelac (2.7). (Author) 76 refs. , 17 tabs. , 4 figs.

Sponge cell culture? A molecular identification method for sponge cells

NARCIS (Netherlands)

Sipkema, D.; Heilig, G.H.J.; Akkermans, A.D.L.; Osinga, R.; Tramper, J.; Wijffels, R.H.

2003-01-01

Dissociated sponge cells are easily confused with unicellular organisms. This has been an obstacle in the development of sponge-cell lines. We developed a molecular detection method to identify cells of the sponge Dysidea avara in dissociated cell cultures. The 18S ribosomal RNA gene from a Dysidea

Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure.

Science.gov (United States)

Akahane, M; Shimizu, T; Kira, T; Onishi, T; Uchihara, Y; Imamura, T; Tanaka, Y

2016-11-01

To assess the structure and extracellular matrix molecule expression of osteogenic cell sheets created via culture in medium with both dexamethasone (Dex) and ascorbic acid phosphate (AscP) compared either Dex or AscP alone. Osteogenic cell sheets were prepared by culturing rat bone marrow stromal cells in a minimal essential medium (MEM), MEM with AscP, MEM with Dex, and MEM with Dex and AscP (Dex/AscP). The cell number and messenger (m)RNA expression were assessed in vitro, and the appearance of the cell sheets was observed after mechanical retrieval using a scraper. β-tricalcium phosphate (β-TCP) was then wrapped with the cell sheets from the four different groups and subcutaneously implanted into rats. After mechanical retrieval, the osteogenic cell sheets from the MEM, MEM with AscP, and MEM with Dex groups appeared to be fragmented or incomplete structures. The cell sheets cultured with Dex/AscP remained intact after mechanical retrieval, without any identifiable tears. Culture with Dex/AscP increased the mRNA and protein expression of extracellular matrix proteins and cell number compared with those of the other three groups. More bridging bone formation was observed after transplantation of the β-TCP scaffold wrapped with cell sheets cultured with Dex/AscP, than in the other groups. These results suggest that culture with Dex/AscP improves the mechanical integrity of the osteogenic cell sheets, allowing retrieval of the confluent cells in a single cell sheet structure. This method may be beneficial when applied in cases of difficult tissue reconstruction, such as nonunion, bone defects, and osteonecrosis.Cite this article: M. Akahane, T. Shimizu, T. Kira, T. Onishi, Y. Uchihara, T. Imamura, Y. Tanaka. Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure. Bone Joint Res 2016;5:569-576. DOI: 10.1302/2046-3758.511.BJR-2016-0013.R1. © 2016 Akahane et al.

21st Century Cell Culture for 21st Century Toxicology.

Science.gov (United States)

Pamies, David; Hartung, Thomas

2017-01-17

There is no good science in bad models. Cell culture is especially prone to artifacts. A number of novel cell culture technologies have become more broadly available in the 21st century, which allow overcoming limitations of traditional culture and are more physiologically relevant. These include the use of stem-cell derived human cells, cocultures of different cell types, scaffolds and extracellular matrices, perfusion platforms (such as microfluidics), 3D culture, organ-on-chip technologies, tissue architecture, and organ functionality. The physiological relevance of such models is further enhanced by the measurement of biomarkers (e.g., key events of pathways), organ specific functionality, and more comprehensive assessment cell responses by high-content methods. These approaches are still rarely combined to create microphysiological systems. The complexity of the combination of these technologies can generate results closer to the in vivo situation but increases the number of parameters to control, bringing some new challenges. In fact, we do not argue that all cell culture needs to be that sophisticated. The efforts taken are determined by the purpose of our experiments and tests. If only a very specific molecular target to cell response is of interest, a very simple model, which reflects this, might be much more suited to allow standardization and high-throughput. However, the less defined the end point of interest and cellular response are, the better we should approximate organ- or tissue-like culture conditions to make physiological responses more probable. Besides these technologic advances, important progress in the quality assurance and reporting on cell cultures as well as the validation of cellular test systems brings the utility of cell cultures to a new level. The advancement and broader implementation of Good Cell Culture Practice (GCCP) is key here. In toxicology, this is a major prerequisite for meaningful and reliable results, ultimately

Microfluidic perfusion culture of human induced pluripotent stem cells under fully defined culture conditions.

Science.gov (United States)

Yoshimitsu, Ryosuke; Hattori, Koji; Sugiura, Shinji; Kondo, Yuki; Yamada, Rotaro; Tachikawa, Saoko; Satoh, Taku; Kurisaki, Akira; Ohnuma, Kiyoshi; Asashima, Makoto; Kanamori, Toshiyuki

2014-05-01

Human induced pluripotent stem cells (hiPSCs) are a promising cell source for drug screening. For this application, self-renewal or differentiation of the cells is required, and undefined factors in the culture conditions are not desirable. Microfluidic perfusion culture allows the production of small volume cultures with precisely controlled microenvironments, and is applicable to high-throughput cellular environment screening. Here, we developed a microfluidic perfusion culture system for hiPSCs that uses a microchamber array chip under defined extracellular matrix (ECM) and culture medium conditions. By screening various ECMs we determined that fibronectin and laminin are appropriate for microfluidic devices made out of the most popular material, polydimethylsiloxane (PDMS). We found that the growth rate of hiPSCs under pressure-driven perfusion culture conditions was higher than under static culture conditions in the microchamber array. We applied our new system to self-renewal and differentiation cultures of hiPSCs, and immunocytochemical analysis showed that the state of the hiPSCs was successfully controlled. The effects of three antitumor drugs on hiPSCs were comparable between microchamber array and 96-well plates. We believe that our system will be a platform technology for future large-scale screening of fully defined conditions for differentiation cultures on integrated microfluidic devices. © 2013 Wiley Periodicals, Inc.

Quantitative volumetric Raman imaging of three dimensional cell cultures

Science.gov (United States)

Kallepitis, Charalambos; Bergholt, Mads S.; Mazo, Manuel M.; Leonardo, Vincent; Skaalure, Stacey C.; Maynard, Stephanie A.; Stevens, Molly M.

2017-03-01

The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell-material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.

The Effect of Primary Cancer Cell Culture Models on the Results of Drug Chemosensitivity Assays: The Application of Perfusion Microbioreactor System as Cell Culture Vessel

Science.gov (United States)

Chen, Yi-Dao; Huang, Shiang-Fu; Wang, Hung-Ming

2015-01-01

To precisely and faithfully perform cell-based drug chemosensitivity assays, a well-defined and biologically relevant culture condition is required. For the former, a perfusion microbioreactor system capable of providing a stable culture condition was adopted. For the latter, however, little is known about the impact of culture models on the physiology and chemosensitivity assay results of primary oral cavity cancer cells. To address the issues, experiments were performed. Results showed that minor environmental pH change could significantly affect the metabolic activity of cells, demonstrating the importance of stable culture condition for such assays. Moreover, the culture models could also significantly influence the metabolic activity and proliferation of cells. Furthermore, the choice of culture models might lead to different outcomes of chemosensitivity assays. Compared with the similar test based on tumor-level assays, the spheroid model could overestimate the drug resistance of cells to cisplatin, whereas the 2D and 3D culture models might overestimate the chemosensitivity of cells to such anticancer drug. In this study, the 3D culture models with same cell density as that in tumor samples showed comparable chemosensitivity assay results as the tumor-level assays. Overall, this study has provided some fundamental information for establishing a precise and faithful drug chemosensitivity assay. PMID:25654105

Bags versus flasks: a comparison of cell culture systems for the production of dendritic cell-based immunotherapies.

Science.gov (United States)

Fekete, Natalie; Béland, Ariane V; Campbell, Katie; Clark, Sarah L; Hoesli, Corinne A

2018-04-19

In recent years, cell-based therapies targeting the immune system have emerged as promising strategies for cancer treatment. This review summarizes manufacturing challenges related to production of antigen presenting cells as a patient-tailored cancer therapy. Understanding cell-material interactions is essential because in vitro cell culture manipulations to obtain mature antigen-producing cells can significantly alter their in vivo performance. Traditional antigen-producing cell culture protocols often rely on cell adhesion to surface-treated hydrophilic polystyrene flasks. More recent commercial and investigational cancer immunotherapy products were manufactured using suspension cell culture in closed hydrophobic fluoropolymer bags. The shift to closed cell culture systems can decrease risks of contamination by individual operators, as well as facilitate scale-up and automation. Selecting closed cell culture bags over traditional open culture systems entails different handling procedures and processing controls, which can affect product quality. Changes in culture vessels also entail changes in vessel materials and geometry, which may alter the cell microenvironment and resulting cell fate decisions. Strategically designed culture systems will pave the way for the generation of more sophisticated and highly potent cell-based cancer vaccines. As an increasing number of cell-based therapies enter the clinic, the selection of appropriate cell culture vessels and materials becomes a critical consideration that can impact the therapeutic efficacy of the product, and hence clinical outcomes and patient quality of life. © 2018 The Authors Transfusion published by Wiley Periodicals, Inc. on behalf of AABB.

INFLUENCE OF CULTIVARS AND SEED THERMAL TREATMENT ON THE DEVELOPMENT OF FUNGAL PATHOGENS IN CARROT AND ONION PLANTS

Czech Academy of Sciences Publication Activity Database

Koudela, M.; Novotný, Čeněk

2016-01-01

Roč. 64, č. 4 (2016), s. 1181-1189 ISSN 1211-8516 R&D Projects: GA MZe QJ1210165 Institutional support: RVO:61388971 Keywords : carrot * onion * fungal pathogens * plants infection Subject RIV: EE - Microbiology, Virology

Three-dimensional hydrogel cell culture systems for modeling neural tissue

Science.gov (United States)

Frampton, John

Two-dimensional (2-D) neural cell culture systems have served as physiological models for understanding the cellular and molecular events that underlie responses to physical and chemical stimuli, control sensory and motor function, and lead to the development of neurological diseases. However, the development of three-dimensional (3-D) cell culture systems will be essential for the advancement of experimental research in a variety of fields including tissue engineering, chemical transport and delivery, cell growth, and cell-cell communication. In 3-D cell culture, cells are provided with an environment similar to tissue, in which they are surrounded on all sides by other cells, structural molecules and adhesion ligands. Cells grown in 3-D culture systems display morphologies and functions more similar to those observed in vivo, and can be cultured in such a way as to recapitulate the structural organization and biological properties of tissue. This thesis describes a hydrogel-based culture system, capable of supporting the growth and function of several neural cell types in 3-D. Alginate hydrogels were characterized in terms of their biomechanical and biochemical properties and were functionalized by covalent attachment of whole proteins and peptide epitopes. Methods were developed for rapid cross-linking of alginate hydrogels, thus permitting the incorporation of cells into 3-D scaffolds without adversely affecting cell viability or function. A variety of neural cell types were tested including astrocytes, microglia, and neurons. Cells remained viable and functional for longer than two weeks in culture and displayed process outgrowth in 3-D. Cell constructs were created that varied in cell density, type and organization, providing experimental flexibility for studying cell interactions and behavior. In one set of experiments, 3-D glial-endothelial cell co-cultures were used to model blood-brain barrier (BBB) structure and function. This co-culture system was

Biological Control of Meloidogyne hapla Using an Antagonistic Bacterium

Directory of Open Access Journals (Sweden)

Jiyeong Park

2014-09-01

Full Text Available We examined the efficacy of a bacterium for biocontrol of the root-knot nematode (RKN Meloidogyne hapla in carrot (Daucus carota subsp. sativus and tomato (Solanum lycopersicum. Among 542 bacterial isolates from various soils and plants, the highest nematode mortality was observed for treatments with isolate C1-7, which was identified as Bacillus cereus based on cultural and morphological characteristics, the Biolog program, and 16S rRNA sequencing analyses. The population density and the nematicidal activity of B. cereus C1-7 remained high until the end of culture in brain heart infusion broth, suggesting that it may have sustainable biocontrol potential. In pot experiments, the biocontrol efficacy of B. cereus C1-7 was high, showing complete inhibition of root gall or egg mass formation by RKN in carrot and tomato plants, and subsequently reducing RKN damage and suppressing nematode population growth, respectively. Light microscopy of RKN-infected carrot root tissues treated with C1-7 showed reduced formation of gall cells and fully developed giant cells, while extensive gall cells and fully mature giant cells with prominent cell wall ingrowths formed in the untreated control plants infected with RKNs. These histopathological characteristics may be the result of residual or systemic biocontrol activity of the bacterium, which may coincide with the biocontrol efficacies of nematodes in pots. These results suggest that B. cereus C1-7 can be used as a biocontrol agent for M. hapla.

Cell-cycle distributions and radiation responses of Chinese hamster cells cultured continuously under hypoxic conditions

International Nuclear Information System (INIS)

Tokita, N.; Carpenter, S.G.; Raju, M.R.

1984-01-01

Cell-cycle distributions were measured by flow cytometry for Chinese hamster (CHO) cells cultured continuously under hypoxic conditions. DNA histograms showed an accumulation of cells in the early S phase followed by a traverse delay through the S phase, and a G 2 block. During hypoxic culturing, cell viability decreased rapidly to less than 0.1% at 120 h. Radiation responses for cells cultured under these conditions showed an extreme radioresistance at 72 h. Results suggest that hypoxia induces a condition similar to cell synchrony which itself changes the radioresistance of hypoxic cells. (author)

Engineering systems for the generation of patterned co-cultures for controlling cell-cell interactions.

Science.gov (United States)

Kaji, Hirokazu; Camci-Unal, Gulden; Langer, Robert; Khademhosseini, Ali

2011-03-01

Inside the body, cells lie in direct contact or in close proximity to other cell types in a tightly controlled architecture that often regulates the resulting tissue function. Therefore, tissue engineering constructs that aim to reproduce the architecture and the geometry of tissues will benefit from methods of controlling cell-cell interactions with microscale resolution. We discuss the use of microfabrication technologies for generating patterned co-cultures. In addition, we categorize patterned co-culture systems by cell type and discuss the implications of regulating cell-cell interactions in the resulting biological function of the tissues. Patterned co-cultures are a useful tool for fabricating tissue engineered constructs and for studying cell-cell interactions in vitro, because they can be used to control the degree of homotypic and heterotypic cell-cell contact. In addition, this approach can be manipulated to elucidate important factors involved in cell-matrix interactions. Patterned co-culture strategies hold significant potential to develop biomimetic structures for tissue engineering. It is expected that they would create opportunities to develop artificial tissues in the future. This article is part of a Special Issue entitled Nanotechnologies - Emerging Applications in Biomedicine. 2010 Elsevier B.V. All rights reserved.

The influence of the fungal pathogen Mycocentrospora acerina on the proteome and polyacetylenes and 6-methoxymellein in organic and conventionally cultivated carrots (Daucus carota) during post harvest storage

DEFF Research Database (Denmark)

Louarn, Sébastien; Nawrocki, Arkadiusz; Edelenbos, Merete

2012-01-01

Many carrots are discarded during post harvest cold storage due to development of fungal infections, caused by, e.g., Mycocentrospora acerina (liquorice rot). We compared the susceptibility of carrots grown under conventional and organic agricultural practices. In one year, organically cultivated...

Morphological and Immunohistochemical Characterization of Canine Osteosarcoma Spheroid Cell Cultures.

Science.gov (United States)

Gebhard, C; Gabriel, C; Walter, I

2016-06-01

Spheroid cell culture emerges as powerful in vitro tool for experimental tumour research. In this study, we established a scaffold-free three-dimensional spheroid system built from canine osteosarcoma (OS) cells (D17). Spheroids (7, 14 and 19 days of cultivation) and monolayer cultures (2 and 7 days of cultivation) were evaluated and compared on light and electron microscopy. Monolayer and spheroid cultures were tested for vimentin, cytokeratin, alkaline phosphatase, osteocalcin and collagen I by means of immunohistochemistry. The spheroid cell culture exhibited a distinct network of collagen I in particular after 19-day cultivation, whereas in monolayer cultures, collagen I was arranged as a lamellar basal structure. Necrotic centres of large spheroids, as observed in 14- and 19-day cultures, were characterized by significant amounts of osteocalcin. Proliferative activity as determined by Ki-67 immunoreactivity showed an even distribution in two-dimensional cultures. In spheroids, proliferation was predominating in the peripheral areas. Metastasis-associated markers ezrin and S100A4 were shown to be continuously expressed in monolayer and spheroid cultures. We conclude that the scaffold-free spheroid system from canine OS cells has the ability to mimic the architecture of the in vivo tumour, in particular cell-cell and cell-matrix interactions. © 2015 The Authors. Anatomia, Histologia, Embryologia Published by Blackwell Verlag GmbH.

A microwell cell culture platform for the aggregation of pancreatic β-cells.

Science.gov (United States)

Bernard, Abigail B; Lin, Chien-Chi; Anseth, Kristi S

2012-08-01

Cell-cell contact between pancreatic β-cells is important for maintaining survival and normal insulin secretion. Various techniques have been developed to promote cell-cell contact between β-cells, but a simple yet robust method that affords precise control over three-dimensional (3D) β-cell cluster size has not been demonstrated. To address this need, we developed a poly(ethylene glycol) (PEG) hydrogel microwell platform using photolithography. This microwell cell-culture platform promotes the formation of 3D β-cell aggregates of defined sizes from 25 to 210 μm in diameter. Using this platform, mouse insulinoma 6 (MIN6) β-cells formed aggregates with cell-cell adherin junctions. These naturally formed cell aggregates with controllable sizes can be removed from the microwells for macroencapsulation, implantation, or other biological assays. When removed and subsequently encapsulated in PEG hydrogels, the aggregated cell clusters demonstrated improved cellular viability (>90%) over 7 days in culture, while the β-cells encapsulated as single cells maintained only 20% viability. Aggregated MIN6 cells also exhibited more than fourfold higher insulin secretion in response to a glucose challenge compared with encapsulated single β-cells. Further, the cell aggregates stained positively for E-cadherin, indicative of the formation of cell junctions. Using this hydrogel microwell cell-culture method, viable and functional β-cell aggregates of specific sizes were created, providing a platform from which other biologically relevant questions may be answered.

Quantitative volumetric Raman imaging of three dimensional cell cultures

KAUST Repository

Kallepitis, Charalambos

2017-03-22

The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell–material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.

Cell division requirement for activation of murine leukemia virus in cell culture by irradiation

International Nuclear Information System (INIS)

Otten, J.A.; Quarles, J.M.; Tennant, R.W.

1976-01-01

Actively dividing cultures of AKR mouse cells were exposed to relatively low dose-rates of γ radiation and tested for activation of endogenous leukemia viruses. Efficient and reproducible induction of virus was obtained with actively dividing cells, but cultures deprived of serum to inhibit cell division before and during γ irradiation were not activated, even when medium with serum was added immediately after irradiation. These results show that cell division was required for virus induction but that a stable intermediate similar to the state induced by halogenated pyrimidines was not formed. In actively dividing AKR cell cultures, virus activation appeared to be proportional to the dose of γ radiation; the estimated frequency of activation was 1-8 x 10 - 5 per exposed cell and the efficiency of activation was approximately 0.012 inductions per cell per rad. Other normal primary and established mouse cell cultures tested were not activated by γ radiation. The requirement of cell division for radiation and chemical activation may reflect some common mechanism for initiation of virus expression

T cell resistance to activation by dendritic cells requires long-term culture in simulated microgravity

Science.gov (United States)

Bradley, Jillian H.; Stein, Rachel; Randolph, Brad; Molina, Emily; Arnold, Jennifer P.; Gregg, Randal K.

2017-11-01

Immune impairment mediated by microgravity threatens the success of space exploration requiring long-duration spaceflight. The cells of most concern, T lymphocytes, coordinate the host response against microbial and cancerous challenges leading to elimination and long-term protection. T cells are activated upon recognition of specific microbial peptides bound on the surface of antigen presenting cells, such as dendritic cells (DC). Subsequently, this engagement results in T cell proliferation and differentiation into effector T cells driven by autocrine interleukin-2 (IL-2) and other cytokines. Finally, the effector T cells acquire the weaponry needed to destroy microbial invaders and tumors. Studies conducted on T cells during spaceflight, or using Earth-based culture systems, have shown reduced production of cytokines, proliferation and effector functions as compared to controls. This may account for the cases of viral reactivation events and opportunistic infections associated with astronauts of numerous missions. This work has largely been based upon the outcome of T cell activation by stimulatory factors that target select T cell signaling pathways rather than the complex, signaling events related to the natural process of antigen presentation by DC. This study tested the response of an ovalbumin peptide-specific T cell line, OT-II TCH, to activation by DC when the T cells were cultured 24-120 h in a simulated microgravity (SMG) environment generated by a rotary cell culture system. Following 72 h culture of T cells in SMG (SMG-T) or control static (Static-T) conditions, IL-2 production by the T cells was reduced in SMG-T cells compared to Static-T cells upon stimulation by phorbol 12-myristate 13-acetate (PMA) and ionomycin. However, when the SMG-T cells were stimulated with DC and peptide, IL-2 was significantly increased compared to Static-T cells. Such enhanced IL-2 production by SMG-T cells peaked at 72 h SMG culture time and decreased thereafter. When

Tumor necrosis factor (cachetin) decreases adipose cell differentiation in primary cell culture

International Nuclear Information System (INIS)

Martin, R.J.; Jones, D.D.; Jewell, D.E.; Hausman, G.J.

1986-01-01

Cachetin has been shown to effect gene product expression in the established adipose cell line 3T3-L1. Expression of messenger RNA for lipoprotein lipase is suppressed in cultured adipocytes. The purpose of this study was to determine the effect of Cachetin on adipose cell differentiation in primary cell culture. Stromalvascular cells obtained from the inguinal fat pad of 4-5 week old Sprague-Dawley rats were grown in culture for two weeks. During the proliferative growth phase all cells were grown on the same medium and labelled with 3 H-thymidine. Cachetin treatment (10 -6 to 10 -10 M) was initiated on day 5, the initial phase of preadipocyte differentiation. Adipocytes and stromal cells were separated using density gradient, and 3 H-thymidine was determined for both cell types. Thymidine incorporation into adipose cells was decreased maximally (∼ 50%) at 10 -10 M. Stromalvascular cells were not influenced at any of the doses tested. Adipose cell lipid content as indicated by oil red-O staining was decreased by Cachetin. Esterase staining by adipose cells treated with Cachetin was increased indicating an increase in intracellular lipase. These studies show that Cachetin has specific effects on primary adipose cell differentiation

Feeding Frequency Affects Cultured Rat Pituitary Cells in Low Gravity

Science.gov (United States)

Hymer, W. C.; Grindeland, R. E.; Salada, T.; Cenci, R.; Krishnan, K.; Mukai, C.; Nagaoka, S.

1996-01-01

In this report, we describe the results of a rat pituitary cell culture experiment done on STS-65 in which the effect of cell feeding on the release of the six anterior pituitary hormones was studied. We found complex microgravity related interactions between the frequency of cell feeding and the quantity and quality (i.e. biological activity) of some of the six hormones released in flight. Analyses of growth hormone (GH) released from cells into culture media on different mission days using gel filtration and ion exchange chromatography yielded qualitatively similar results between ground and flight samples. Lack of cell feeding resulted in extensive cell clumping in flight (but not ground) cultures. Vigorous fibroblast growth occurred in both ground and flight cultures fed 4 times. These results are interpreted within the context of autocrine and or paracrine feedback interactions. Finally the payload specialist successfully prepared a fresh trypsin solution in microgravity, detached the cells from their surface and reinserted them back into the culture chamber. These cells reattached and continued to release hormone in microgravity. In summary, this experiment shows that pituitary cells are microgravity sensitive and that coupled operations routinely associated with laboratory cel1 culture can also be accomplished in low gravity.

Utilization of Landsat-8 data for the estimation of carrot and maize crop water footprint under the arid climate of Saudi Arabia.

Science.gov (United States)

Madugundu, Rangaswamy; Al-Gaadi, Khalid A; Tola, ElKamil; Hassaballa, Abdalhaleem A; Kayad, Ahmed G

2018-01-01

The crop Water Footprint (WF) can provide a comprehensive knowledge of the use of water through the demarcation of the amount of the water consumed by different crops. The WF has three components: green (WFg), blue (WFb) and grey (WFgr) water footprints. The WFg refers to the rainwater stored in the root zone soil layer and is mainly utilized for agricultural, horticultural and forestry production. The WFb, however, is the consumptive use of water from surface or groundwater resources and mainly deals with irrigated agriculture, industry, domestic water use, etc. While the WFgr is the amount of fresh water required to assimilate pollutants resulting from the use of fertilizers/agrochemicals. This study was conducted on six agricultural fields in the Eastern region of Saudi Arabia, during the period from December 2015 to December 2016, to investigate the spatiotemporal variation of the WF of silage maize and carrot crops. The WF of each crop was estimated in two ways, namely agro-meteorological (WFAgro) and remote sensing (WFRS) methods. The blue, green and grey components of WFAgro were computed with the use of weather station/Eddy covariance measurements and field recorded crop yield datasets. The WFRS estimated by applying surface energy balance principles on Landsat-8 imageries. However, due to non-availability of Landsat-8 data on the event of rainy days, this study was limited to blue component (WFRS-b). The WFAgro of silage maize was found to range from 3545 m3 t-1 to 4960 m3 t-1; on an average, the WFAgro-g, WFAgro-b, and WFAgro-gr are composed of < 1%, 77%, and 22%, respectively. In the case of carrot, the WFAgro ranged between 297 m3 t-1 and 502 m3 t-1. The WFAgro-g of carrot crop was estimated at <1%, while WFAgro-b and WFAgro-gr was 67% and 32%, respectively. The WFAgro-b is occupied as a major portion in WF of silage maize (77%) and carrot (68%) crops. This is due to the high crop water demand combined with a very erratic rainfall, the irrigation is

Utilization of Landsat-8 data for the estimation of carrot and maize crop water footprint under the arid climate of Saudi Arabia.

Directory of Open Access Journals (Sweden)

Rangaswamy Madugundu

Full Text Available The crop Water Footprint (WF can provide a comprehensive knowledge of the use of water through the demarcation of the amount of the water consumed by different crops. The WF has three components: green (WFg, blue (WFb and grey (WFgr water footprints. The WFg refers to the rainwater stored in the root zone soil layer and is mainly utilized for agricultural, horticultural and forestry production. The WFb, however, is the consumptive use of water from surface or groundwater resources and mainly deals with irrigated agriculture, industry, domestic water use, etc. While the WFgr is the amount of fresh water required to assimilate pollutants resulting from the use of fertilizers/agrochemicals. This study was conducted on six agricultural fields in the Eastern region of Saudi Arabia, during the period from December 2015 to December 2016, to investigate the spatiotemporal variation of the WF of silage maize and carrot crops. The WF of each crop was estimated in two ways, namely agro-meteorological (WFAgro and remote sensing (WFRS methods. The blue, green and grey components of WFAgro were computed with the use of weather station/Eddy covariance measurements and field recorded crop yield datasets. The WFRS estimated by applying surface energy balance principles on Landsat-8 imageries. However, due to non-availability of Landsat-8 data on the event of rainy days, this study was limited to blue component (WFRS-b. The WFAgro of silage maize was found to range from 3545 m3 t-1 to 4960 m3 t-1; on an average, the WFAgro-g, WFAgro-b, and WFAgro-gr are composed of < 1%, 77%, and 22%, respectively. In the case of carrot, the WFAgro ranged between 297 m3 t-1 and 502 m3 t-1. The WFAgro-g of carrot crop was estimated at <1%, while WFAgro-b and WFAgro-gr was 67% and 32%, respectively. The WFAgro-b is occupied as a major portion in WF of silage maize (77% and carrot (68% crops. This is due to the high crop water demand combined with a very erratic rainfall, the

Carrot Juice Administration Decreases Liver Stearoyl-CoA Desaturase 1 and Improves Docosahexaenoic Acid Levels, but Not Steatosis in High Fructose Diet-Fed Weanling Wistar Rats.

Science.gov (United States)

Mahesh, Malleswarapu; Bharathi, Munugala; Reddy, Mooli Raja Gopal; Kumar, Manchiryala Sravan; Putcha, Uday Kumar; Vajreswari, Ayyalasomayajula; Jeyakumar, Shanmugam M

2016-09-01

Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent liver diseases associated with an altered lifestyle, besides genetic factors. The control and management of NAFLD mostly depend on lifestyle modifications, due to the lack of a specific therapeutic approach. In this context, we assessed the effect of carrot juice on the development of high fructose-induced hepatic steatosis. For this purpose, male weanling Wistar rats were divided into 4 groups, fed either a control (Con) or high fructose (HFr) diet of AIN93G composition, with or without carrot juice (CJ) for 8 weeks. At the end of the experimental period, plasma biochemical markers, such as triglycerides, alanine aminotransferase, and β-hydroxy butyrate levels were comparable among the 4 groups. Although, the liver injury marker, aspartate aminotransferase, levels in plasma showed a reduction, hepatic triglycerides levels were not significantly reduced by carrot juice ingestion in the HFr diet-fed rats (HFr-CJ). On the other hand, the key triglyceride synthesis pathway enzyme, hepatic stearoyl-CoA desaturase 1 (SCD1), expression at mRNA level was augmented by carrot juice ingestion, while their protein levels showed a significant reduction, which corroborated with decreased monounsaturated fatty acids (MUFA), particularly palmitoleic (C16:1) and oleic (C18:1) acids. Notably, it also improved the long chain n-3 polyunsaturated fatty acid, docosahexaenoic acid (DHA; C22:6) content of the liver in HFr-CJ. In conclusion, carrot juice ingestion decreased the SCD1-mediated production of MUFA and improved DHA levels in liver, under high fructose diet-fed conditions. However, these changes did not significantly lower the hepatic triglyceride levels.

Shelf-life extension and improving micro-biological quality of mixed peas with diced carrot by gamma irradiation

International Nuclear Information System (INIS)

Taha, S. M.; Hammad, A. A.; Amal, S. M.; Gebreel, H. M.

2010-01-01

Mixed peas with diced carrot were collected and examined for their microbiological quality. All the examined samples had high level of microbial load. All examined samples contained Escherichia coli (E. coli) and Enterococcus faecalis (Ent. faecalis). Staphylococcus aureus (S. aureus) was detected in only 4 of samples (26.3%). The tested samples were free from Aeromonas hydrophila (A. hydrophila), Listeria monocytogenes (L monocytogenes) and Salmonella species. Gamma irradiation caused a great reduction in all microbial loads. During refrigerated storage, the counts of all microorganisms increased, but the rate of increase was slower as the irradiation dose increased. Irradiation dose of 3 kGy was the optimum dose for preservation of mixed peas with diced carrot which extended the refrigeration shelf-life up to 21 days and it was sufficient in eliminating pathogenic bacteria without affecting their sensory quality and with negligible effect on chemical quality.

Characterization of primary human mammary epithelial cells isolated and propagated by conditional reprogrammed cell culture.

Science.gov (United States)

Jin, Liting; Qu, Ying; Gomez, Liliana J; Chung, Stacey; Han, Bingchen; Gao, Bowen; Yue, Yong; Gong, Yiping; Liu, Xuefeng; Amersi, Farin; Dang, Catherine; Giuliano, Armando E; Cui, Xiaojiang

2018-02-20

Conditional reprogramming methods allow for the inexhaustible in vitro proliferation of primary epithelial cells from human tissue specimens. This methodology has the potential to enhance the utility of primary cell culture as a model for mammary gland research. However, few studies have systematically characterized this method in generating in vitro normal human mammary epithelial cell models. We show that cells derived from fresh normal breast tissues can be propagated and exhibit heterogeneous morphologic features. The cultures are composed of CK18, desmoglein 3, and CK19-positive luminal cells and vimentin, p63, and CK14-positive myoepithelial cells, suggesting the maintenance of in vivo heterogeneity. In addition, the cultures contain subpopulations with different CD49f and EpCAM expression profiles. When grown in 3D conditions, cells self-organize into distinct structures that express either luminal or basal cell markers. Among these structures, CK8-positive cells enclosing a lumen are capable of differentiation into milk-producing cells in the presence of lactogenic stimulus. Furthermore, our short-term cultures retain the expression of ERα, as well as its ability to respond to estrogen stimulation. We have investigated conditionally reprogrammed normal epithelial cells in terms of cell type heterogeneity, cellular marker expression, and structural arrangement in two-dimensional (2D) and three-dimensional (3D) systems. The conditional reprogramming methodology allows generation of a heterogeneous culture from normal human mammary tissue in vitro . We believe that this cell culture model will provide a valuable tool to study mammary cell function and malignant transformation.

Antitumor Activity of Rat Mesenchymal Stem Cells during Direct or Indirect Co-Culturing with C6 Glioma Cells.

Science.gov (United States)

Gabashvili, A N; Baklaushev, V P; Grinenko, N F; Mel'nikov, P A; Cherepanov, S A; Levinsky, A B; Chehonin, V P

2016-02-01

The tumor-suppressive effect of rat mesenchymal stem cells against low-differentiated rat C6 glioma cells during their direct and indirect co-culturing and during culturing of C6 glioma cells in the medium conditioned by mesenchymal stem cells was studied in an in vitro experiment. The most pronounced antitumor activity of mesenchymal stem cells was observed during direct co-culturing with C6 glioma cells. The number of live C6 glioma cells during indirect co-culturing and during culturing in conditioned medium was slightly higher than during direct co-culturing, but significantly differed from the control (C6 glioma cells cultured in medium conditioned by C6 glioma cells). The cytotoxic effect of medium conditioned by mesenchymal stem cells was not related to medium depletion by glioma cells during their growth. The medium conditioned by other "non-stem" cells (rat astrocytes and fibroblasts) produced no tumor-suppressive effect. Rat mesenchymal stem cells, similar to rat C6 glioma cells express connexin 43, the main astroglial gap junction protein. During co-culturing, mesenchymal stem cells and glioma C6 cells formed functionally active gap junctions. Gap junction blockade with connexon inhibitor carbenoxolone attenuated the antitumor effect observed during direct co-culturing of C6 glioma cells and mesenchymal stem cells to the level produced by conditioned medium. Cell-cell signaling mediated by gap junctions can be a mechanism of the tumor-suppressive effect of mesenchymal stem cells against C6 glioma cells. This phenomenon can be used for the development of new methods of cell therapy for high-grade malignant gliomas.

Advances in tissue engineering through stem cell-based co-culture.

Science.gov (United States)

Paschos, Nikolaos K; Brown, Wendy E; Eswaramoorthy, Rajalakshmanan; Hu, Jerry C; Athanasiou, Kyriacos A

2015-05-01

Stem cells are the future in tissue engineering and regeneration. In a co-culture, stem cells not only provide a target cell source with multipotent differentiation capacity, but can also act as assisting cells that promote tissue homeostasis, metabolism, growth and repair. Their incorporation into co-culture systems seems to be important in the creation of complex tissues or organs. In this review, critical aspects of stem cell use in co-culture systems are discussed. Direct and indirect co-culture methodologies used in tissue engineering are described, along with various characteristics of cellular interactions in these systems. Direct cell-cell contact, cell-extracellular matrix interaction and signalling via soluble factors are presented. The advantages of stem cell co-culture strategies and their applications in tissue engineering and regenerative medicine are portrayed through specific examples for several tissues, including orthopaedic soft tissues, bone, heart, vasculature, lung, kidney, liver and nerve. A concise review of the progress and the lessons learned are provided, with a focus on recent developments and their implications. It is hoped that knowledge developed from one tissue can be translated to other tissues. Finally, we address challenges in tissue engineering and regenerative medicine that can potentially be overcome via employing strategies for stem cell co-culture use. Copyright © 2014 John Wiley & Sons, Ltd.

Stimulation of DNA synthesis in cultured rat alveolar type II cells

International Nuclear Information System (INIS)

Leslie, C.C.; McCormick-Shannon, K.; Robinson, P.C.; Mason, R.J.

1985-01-01

Restoration of the alveolar epithelium after injury is thought to be dependent on the proliferation of alveolar type II cells. To understand the factors that may be involved in promoting type II cell proliferation in vivo, we determined the effect of potential mitogens and culture substrata on DNA synthesis in rat alveolar type II cells in primary culture. Type II cells cultured in basal medium containing 10% fetal bovine serum (FBS) exhibited essentially no DNA synthesis. Factors that stimulated 3 H-thymidine incorporation included cholera toxin, epidermal growth factor, and rat serum. The greatest degree of stimulation was achieved by plating type II cells on an extracellular matrix prepared from bovine corneal endothelial cells and then by culturing the pneumocytes in medium containing rat serum, cholera toxin, insulin, and epidermal growth factor. Under conditions of stimulation of 3 H-thymidine incorporation there was an increased DNA content per culture dish but no increase in cell number. The ability of various culture conditions to promote DNA synthesis in type II cells was verified by autoradiography. Type II cells were identified by the presence of cytoplasmic inclusions, which were visualized by tannic acid staining before autoradiography. These results demonstrate the importance of soluble factors and culture substratum in stimulating DNA synthesis in rat alveolar type II cells in primary culture

EXPLANTATION OF MESANGIAL CELL HILLOCKS - A METHOD FOR OBTAINING HUMAN MESANGIAL CELLS IN CULTURE

NARCIS (Netherlands)

MULLER, EW; KIM, Y; MICHAEL, AF; VERNIER, RL; VANDERHEM, GK; VANDERWOUDE, FJ

A simple method is presented for selective cell culture of human mesangial cells using explanatation of mesangial cell hillocks. Glomeruli which had been incubated with collagenase were explanted on plastic tissue culture flasks. Three to 6 weeks after explantation, a rapidly growing multilayer of

Effects of cell concentrations on the survival and repopulation of haemopoietic stem cells in irradiated bone marrow cell culture in vitro

International Nuclear Information System (INIS)

Fujitake, Hideki; Okamoto, Yuruko; Okubo, Hiroshi; Miyanomae, Takeshi; Kumagai, Keiko; Mori, K.J.

1981-01-01

Effects of cell concentrations on the survival and repopulation of haemopoietic stem cells after irradiation were studied in the long-term culture of mouse bone marrow cells in vitro. No difference was observed in the survival of the stem cells among cultures in which 0 - 10 7 cells were re-inoculated on the adherent cell colonies in the culture flask. Stem cells showed a significant proliferation within 1 week and the number of the stem cells exceeded the control in 3 weeks after irradiation in the cultures with less than 10 6 re-inoculated cells per flask. In contrast, there was a considerable delay in the onset of stem cell proliferation after irradiation in the culture with 10 7 cells per flask. Based on these results, a possibility that a stimulator of stem cell proliferation, released from irradiated stromal cells, is cancelled by an inhibitory factor produced by irradiated or unirradiated haemopoietic cells is postulated. (author)

21 CFR 876.5885 - Tissue culture media for human ex vivo tissue and cell culture processing applications.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Tissue culture media for human ex vivo tissue and cell culture processing applications. 876.5885 Section 876.5885 Food and Drugs FOOD AND DRUG... DEVICES Therapeutic Devices § 876.5885 Tissue culture media for human ex vivo tissue and cell culture...

Retinal pigment epithelium culture;a potential source of retinal stem cells.

Science.gov (United States)

Akrami, Hassan; Soheili, Zahra-Soheila; Khalooghi, Keynoush; Ahmadieh, Hamid; Rezaie-Kanavi, Mojgan; Samiei, Shahram; Davari, Malihe; Ghaderi, Shima; Sanie-Jahromi, Fatemeh

2009-07-01

To establish human retinal pigment epithelial (RPE) cell culture as a source for cell replacement therapy in ocular diseases. Human cadaver globes were used to isolate RPE cells. Each globe was cut into several pieces of a few millimeters in size. After removing the sclera and choroid, remaining tissues were washed in phosphate buffer saline and RPE cells were isolated using dispase enzyme solution and cultured in Dulbecco's Modified Eagle's Medium: Nutrient Mixture F-12 supplemented with 10% fetal calf serum. Primary cultures of RPE cells were established and spheroid colonies related to progenitor/stem cells developed in a number of cultures. The colonies included purely pigmented or mixed pigmented and non-pigmented cells. After multiple cellular passages, several types of photoreceptors and neural-like cells were detected morphologically. Cellular plasticity in RPE cell cultures revealed promising results in terms of generation of stem/progenitor cells from human RPE cells. Whether the spheroids and neural-like retinal cells were directly derived from retinal stem cells or offspring of trans-differentiating or de-differentiating RPE cells remains to be answered.

Enhanced infectivity of bluetongue virus in cell culture by centrifugation.

OpenAIRE

Sundin, D R; Mecham, J O

1989-01-01

The effects of centrifugation of the infection of cell culture with bluetongue virus (BTV) were investigated. Baby hamster kidney cells were infected with BTV with or without centrifugation. Viral antigen was detected by immunofluorescence at 24 h in both centrifuged and noncentrifuged cultures. However, after 24 h of infection, the production of PFU in centrifuged cell cultures was 10- to 20-fold greater than that seen in cultures not centrifuged. In addition, centrifugation enhanced the dir...

Selenium biofortification of broccoli and carrots grown in soil amended with Se-enriched hyperaccumulator Stanleya pinnata

Science.gov (United States)

Amending soils with Se-hyperaccumulator plant derived sources of selenium (Se) may be useful for increasing Se content in food crops in Se-deficient regions of the world. In this study, we evaluated total Se and the different chemical species of Se in broccoli and carrots grown in soils amended with...

Enrichment of skin-derived neural precursor cells from dermal cell populations by altering culture conditions.

Science.gov (United States)

Bayati, Vahid; Gazor, Rohoullah; Nejatbakhsh, Reza; Negad Dehbashi, Fereshteh

2016-01-01

As stem cells play a critical role in tissue repair, their manipulation for being applied in regenerative medicine is of great importance. Skin-derived precursors (SKPs) may be good candidates for use in cell-based therapy as the only neural stem cells which can be isolated from an accessible tissue, skin. Herein, we presented a simple protocol to enrich neural SKPs by monolayer adherent cultivation to prove the efficacy of this method. To enrich neural SKPs from dermal cell populations, we have found that a monolayer adherent cultivation helps to increase the numbers of neural precursor cells. Indeed, we have cultured dermal cells as monolayer under serum-supplemented (control) and serum-supplemented culture, followed by serum free cultivation (test) and compared. Finally, protein markers of SKPs were assessed and compared in both experimental groups and differentiation potential was evaluated in enriched culture. The cells of enriched culture concurrently expressed fibronectin, vimentin and nestin, an intermediate filament protein expressed in neural and skeletal muscle precursors as compared to control culture. In addition, they possessed a multipotential capacity to differentiate into neurogenic, glial, adipogenic, osteogenic and skeletal myogenic cell lineages. It was concluded that serum-free adherent culture reinforced by growth factors have been shown to be effective on proliferation of skin-derived neural precursor cells (skin-NPCs) and drive their selective and rapid expansion.

T cell resistance to activation by dendritic cells requires long-term culture in simulated microgravity.

Science.gov (United States)

Bradley, Jillian H; Stein, Rachel; Randolph, Brad; Molina, Emily; Arnold, Jennifer P; Gregg, Randal K

2017-11-01

Immune impairment mediated by microgravity threatens the success of space exploration requiring long-duration spaceflight. The cells of most concern, T lymphocytes, coordinate the host response against microbial and cancerous challenges leading to elimination and long-term protection. T cells are activated upon recognition of specific microbial peptides bound on the surface of antigen presenting cells, such as dendritic cells (DC). Subsequently, this engagement results in T cell proliferation and differentiation into effector T cells driven by autocrine interleukin-2 (IL-2) and other cytokines. Finally, the effector T cells acquire the weaponry needed to destroy microbial invaders and tumors. Studies conducted on T cells during spaceflight, or using Earth-based culture systems, have shown reduced production of cytokines, proliferation and effector functions as compared to controls. This may account for the cases of viral reactivation events and opportunistic infections associated with astronauts of numerous missions. This work has largely been based upon the outcome of T cell activation by stimulatory factors that target select T cell signaling pathways rather than the complex, signaling events related to the natural process of antigen presentation by DC. This study tested the response of an ovalbumin peptide-specific T cell line, OT-II TCH, to activation by DC when the T cells were cultured 24-120 h in a simulated microgravity (SMG) environment generated by a rotary cell culture system. Following 72 h culture of T cells in SMG (SMG-T) or control static (Static-T) conditions, IL-2 production by the T cells was reduced in SMG-T cells compared to Static-T cells upon stimulation by phorbol 12-myristate 13-acetate (PMA) and ionomycin. However, when the SMG-T cells were stimulated with DC and peptide, IL-2 was significantly increased compared to Static-T cells. Such enhanced IL-2 production by SMG-T cells peaked at 72 h SMG culture time and decreased thereafter

Carryover of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) from soil to plant and distribution to the different plant compartments studied in cultures of carrots (Daucus carota ssp. Sativus), potatoes (Solanum tuberosum), and cucumbers (Cucumis Sativus).

Science.gov (United States)

Lechner, Mareike; Knapp, Holger

2011-10-26

A vegetation study was carried out to investigate the carryover of Perfluorooctanoic Acid (PFOA) and Perfluorooctane Sulfonate (PFOS) from soil mixed with contaminated sewage sludge to potato, carrot, and cucumber plants. Analysis was done by liquid-extraction using acetonitrile with dispersive SPE cleanup and subsequent HPLC-MS/MS. In order to assess the transfer potential from soil, transfer factors (TF) were calculated for the different plant compartments: TF = [PFC](plant (wet substance))/[PFC](soil (dry weight)). The highest TF were found for the vegetative plant compartments with average values for PFOS below those for PFOA: cucumber, 0.17 (PFOS), 0.88 (PFOA); potato, 0.36 (PFOS), 0.40 (PFOA); carrot, 0.38 (PFOS), 0.53 (PFOA). Transfer of PFOA and PFOS into potato peelings (average values of TF: PFOA 0.03, PFOS 0.04) exceeded the carryover to the peeled tubers (PFOA 0.01, PFOS potatoes (TF < 0.01). For PFOA, it was higher (TF: 0.03).

Identification of a population of cells with hematopoietic stem cell properties in mouse aorta-gonad-mesonephros cultures

International Nuclear Information System (INIS)

Nobuhisa, Ikuo; Ohtsu, Naoki; Okada, Seiji; Nakagata, Naomi; Taga, Tetsuya

2007-01-01

The aorta-gonad-mesonephros (AGM) region is a primary source of definitive hematopoietic cells in the midgestation mouse embryo. In cultures of dispersed AGM regions, adherent cells containing endothelial cells are observed first, and then non-adherent hematopoietic cells are produced. Here we report on the characterization of hematopoietic cells that emerge in the AGM culture. Based on the expression profiles of CD45 and c-Kit, we defined three cell populations: CD45 low c-Kit + cells that had the ability to form hematopoietic cell colonies in methylcellulose media and in co-cultures with stromal cells; CD45 low c-Kit - cells that showed a granulocyte morphology; CD45 high c-Kit low/- that exhibited a macrophage morphology. In co-cultures of OP9 stromal cells and freshly prepared AGM cultures, CD45 low c-Kit + cells from the AGM culture had the abilities to reproduce CD45 low c-Kit + cells and differentiate into CD45 low c-Kit - and CD45 high c-Kit low/- cells, whereas CD45 low c-Kit - and CD45 high c-Kit low/- did not produce CD45 low c-Kit + cells. Furthermore, CD45 low c-Kit + cells displayed a long-term repopulating activity in adult hematopoietic tissue when transplanted into the liver of irradiated newborn mice. These results indicate that CD45 low c-Kit + cells from the AGM culture have the potential to reconstitute multi-lineage hematopoietic cells

Cell fiber-based three-dimensional culture system for highly efficient expansion of human induced pluripotent stem cells.

Science.gov (United States)

Ikeda, Kazuhiro; Nagata, Shogo; Okitsu, Teru; Takeuchi, Shoji

2017-06-06

Human pluripotent stem cells are a potentially powerful cellular resource for application in regenerative medicine. Because such applications require large numbers of human pluripotent stem cell-derived cells, a scalable culture system of human pluripotent stem cell needs to be developed. Several suspension culture systems for human pluripotent stem cell expansion exist; however, it is difficult to control the thickness of cell aggregations in these systems, leading to increased cell death likely caused by limited diffusion of gases and nutrients into the aggregations. Here, we describe a scalable culture system using the cell fiber technology for the expansion of human induced pluripotent stem (iPS) cells. The cells were encapsulated and cultured within the core region of core-shell hydrogel microfibers, resulting in the formation of rod-shaped or fiber-shaped cell aggregations with sustained thickness and high viability. By encapsulating the cells with type I collagen, we demonstrated a long-term culture of the cells by serial passaging at a high expansion rate (14-fold in four days) while retaining its pluripotency. Therefore, our culture system could be used for large-scale expansion of human pluripotent stem cells for use in regenerative medicine.

Cystine uptake by cultured cells originating from dog proximal tubule segments

International Nuclear Information System (INIS)

States, B.; Reynolds, R.; Lee, J.; Segal, S.

1990-01-01

Large numbers of kidney epithelial cells were cultured successfully from isolated dog proximal tubule segments. Cells in primary culture and in first passage retained the cystine-dibasic amino acid co-transporter system which is found in vivo and in freshly isolated proximal tubule segments. In contrast to other cultured cells, the cystine-glutamate anti-porter was absent in primary cultures. However, this anti-porter system seemed to be developing in cells in first passage. The intracellular ratio of cysteine:reduced glutathione (CSH:GSH) was maintained at 1:36 in both primary cultures and in low passage cells. Incubation of cells in primary culture for 5 min at 37 degrees C with 0.025 mM [ 35 S]L-cystine resulted in incorporation of approximately 36 and 8.5% of the label into intracellular CSH and GSH, respectively. These cultured cells, therefore, seem to be an excellent model system for the eventual elucidation of (a) the inticacies of cystine metabolism and (b) regulation of (1) the cystine-dibasic amino acid co-transporter system and (2) the development of the cysteine-glutamate anti-porter system

Inhibition of apoptosis using exosomes in Chinese hamster ovary cell culture.

Science.gov (United States)

Han, Seora; Rhee, Won Jong

2018-05-01

Animal cell culture technology for therapeutic protein production has shown significant improvement over the last few decades. Chinese hamster ovary (CHO) cells have been widely adapted for the production of biopharmaceutical drugs. In the biopharmaceutical industry, it is crucial to develop cell culture media and culturing conditions to achieve the highest productivity and quality. However, CHO cells are significantly affected by apoptosis in the bioreactors, resulting in a substantial decrease in product quantity and quality. Thus, to overcome the obstacle of apoptosis in CHO cell culture, it is critical to develop a novel method that does not have minimal concern of safety or cost. Herein, we showed for the first time that exosomes, which are nano-sized extracellular vesicles, derived from CHO cells inhibited apoptosis in CHO cell culture when supplemented to the culture medium. Flow cytometric and microscopic analyses revealed that substantial amounts of exosomes were delivered to CHO cells. Higher cell viability after staurosporine treatment was observed by exosome supplementation (67.3%) as compared to control (41.1%). Furthermore, exosomes prevented the mitochondrial membrane potential loss and caspase-3 activation, meaning that the exosomes enhanced cellular activities under pro-apoptotic condition. As the exosomes supplements are derived from CHO cells themselves, it is not only beneficial for the biopharmaceutical productivity of CHO cell culture to inhibit apoptosis, but also from a regulatory standpoint to diminish any safety concerns. Thus, we conclude that the method developed in this research may contribute to the biopharmaceutical industry where minimizing apoptosis in CHO cell culture is beneficial. © 2018 Wiley Periodicals, Inc.

Isolation and Characterization of Poliovirus in Cell Culture Systems.

Science.gov (United States)

Thorley, Bruce R; Roberts, Jason A

2016-01-01

The isolation and characterization of enteroviruses by cell culture was accepted as the "gold standard" by clinical virology laboratories. Methods for the direct detection of all enteroviruses by reverse transcription polymerase chain reaction, targeting a conserved region of the genome, have largely supplanted cell culture as the principal diagnostic procedure. However, the World Health Organization's Global Polio Eradication Initiative continues to rely upon cell culture to isolate poliovirus due to the lack of a reliable sensitive genetic test for direct typing of enteroviruses from clinical specimens. Poliovirus is able to infect a wide range of mammalian cell lines, with CD155 identified as the primary human receptor for all three seroytpes, and virus replication leads to an observable cytopathic effect. Inoculation of cell lines with extracts of clinical specimens and subsequent passaging of the cells leads to an increased virus titre. Cultured isolates of poliovirus are suitable for testing by a variety of methods and remain viable for years when stored at low temperature.This chapter describes general procedures for establishing a cell bank and routine passaging of cell lines. While the sections on specimen preparation and virus isolation focus on poliovirus, the protocols are suitable for other enteroviruses.

SAHA-induced TRAIL-sensitisation of Multiple Myeloma cells is enhanced in 3D cell culture.

Science.gov (United States)

Arhoma, A; Chantry, A D; Haywood-Small, S L; Cross, N A

2017-11-15

Multiple Myeloma (MM) is currently incurable despite many novel therapies. Tumour Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) is a potential anti-tumour agent although effects as a single agent are limited. In this study, we investigated whether the Histone Deacetylase (HDAC) inhibitor SAHA can enhance TRAIL-induced apoptosis and target TRAIL resistance in both suspension culture, and 3D cell culture as a model of disseminated MM lesions that form in bone. The effects of SAHA and/or TRAIL in 6 Multiple Myeloma cell lines were assessed in both suspension cultures and in an Alginate-based 3D cell culture model. The effect of SAHA and/or TRAIL was assessed on apoptosis by assessment of nuclear morphology using Hoechst 33342/Propidium Iodide staining. Viable cell number was assessed by CellTiter-Glo luminescence assay, Caspase-8 and -9 activities were measured by Caspase-Glo™ assay kit. TRAIL-resistant cells were generated by culture of RPMI 8226 and NCI-H929 by acute exposure to TRAIL followed by selection of TRAIL-resistant cells. TRAIL significantly induced apoptosis in a dose-dependent manner in OPM-2, RPMI 8226, NCI-H929, U266, JJN-3 MM cell lines and ADC-1 plasma cell leukaemia cells. SAHA amplified TRAIL responses in all lines except OPM-2, and enhanced TRAIL responses were both via Caspase-8 and -9. SAHA treatment induced growth inhibition that further increased in the combination treatment with TRAIL in MM cells. The co-treatment of TRAIL and SAHA reduced viable cell numbers all cell lines. TRAIL responses were further potentiated by SAHA in 3D cell culture in NCI-H929, RPMI 8226 and U266 at lower TRAIL + SAHA doses than in suspension culture. However TRAIL responses in cells that had been selected for TRAIL resistance were not further enhanced by SAHA treatment. SAHA is a potent sensitizer of TRAIL responses in both TRAIL sensitive and resistant cell lines, in both suspension and 3D culture, however SAHA did not sensitise TRAIL-sensitive cell

Flux analysis of mammalian cell culture

NARCIS (Netherlands)

Martens, D.E.; Tramper, J.

2010-01-01

Animal cells are used for the production of vaccines and pharmaceutical proteins. The increase in demand for these products requires an increase in volumetric productivity of animal cell culture processes, which can be attained through an increase in biomass concentration and/or specific

Plant cell culture initiation

NARCIS (Netherlands)

Hall, R.D.

2000-01-01

The use of cultured plant cells in either organized or unorganized form has increased vey considerably in the last 10-15 yr. Many new technologies have been developed and applications in both fundamental and applied research have led to the development of some powerful tools for improving our

Differentiation of Human Pluripotent Stem Cells into Functional Endothelial Cells in Scalable Suspension Culture

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Ruth Olmer

2018-05-01

Full Text Available Summary: Endothelial cells (ECs are involved in a variety of cellular responses. As multifunctional components of vascular structures, endothelial (progenitor cells have been utilized in cellular therapies and are required as an important cellular component of engineered tissue constructs and in vitro disease models. Although primary ECs from different sources are readily isolated and expanded, cell quantity and quality in terms of functionality and karyotype stability is limited. ECs derived from human induced pluripotent stem cells (hiPSCs represent an alternative and potentially superior cell source, but traditional culture approaches and 2D differentiation protocols hardly allow for production of large cell numbers. Aiming at the production of ECs, we have developed a robust approach for efficient endothelial differentiation of hiPSCs in scalable suspension culture. The established protocol results in relevant numbers of ECs for regenerative approaches and industrial applications that show in vitro proliferation capacity and a high degree of chromosomal stability. : In this article, U. Martin and colleagues show the generation of hiPSC endothelial cells in scalable cultures in up to 100 mL culture volume. The generated ECs show in vitro proliferation capacity and a high degree of chromosomal stability after in vitro expansion. The established protocol allows to generate hiPSC-derived ECs in relevant numbers for regenerative approaches. Keywords: hiPSC differentiation, endothelial cells, scalable culture

Electrospinning PCL Scaffolds Manufacture for Three-Dimensional Breast Cancer Cell Culture

Directory of Open Access Journals (Sweden)

Marc Rabionet

2017-08-01

Full Text Available In vitro cell culture is traditionally performed within two-dimensional (2D environments, providing a quick and cheap way to study cell properties in a laboratory. However, 2D systems differ from the in vivo environment and may not mimic the physiological cell behavior realistically. For instance, 2D culture models are thought to induce cancer stem cells (CSCs differentiation, a rare cancer cell subpopulation responsible for tumor initiation and relapse. This fact hinders the development of therapeutic strategies for tumors with a high relapse percentage, such as triple negative breast cancer (TNBC. Thus, three-dimensional (3D scaffolds have emerged as an attractive alternative to monolayer culture, simulating the extracellular matrix structure and maintaining the differentiation state of cells. In this work, scaffolds were fabricated through electrospinning different poly(ε-caprolactone-acetone solutions. Poly(ε-caprolactone (PCL meshes were seeded with triple negative breast cancer (TNBC cells and 15% PCL scaffolds displayed significantly (p < 0.05 higher cell proliferation and elongation than the other culture systems. Moreover, cells cultured on PCL scaffolds exhibited higher mammosphere forming capacity and aldehyde dehydrogenase activity than 2D-cultured cells, indicating a breast CSCs enrichment. These results prove the powerful capability of electrospinning technology in terms of poly(ε-caprolactone nanofibers fabrication. In addition, this study has demonstrated that electrospun 15% PCL scaffolds are suitable tools to culture breast cancer cells in a more physiological way and to expand the niche of breast CSCs. In conclusion, three-dimensional cell culture using PCL scaffolds could be useful to study cancer stem cell behavior and may also trigger the development of new specific targets against such malignant subpopulation.

Effect of the gamma radiation of cobalt 60 on the beta carotids present in the carrot

International Nuclear Information System (INIS)

Perez Lopez, Sergio Victor Hugo

1997-01-01

In the present work it was investigated the effect of the gamma radiation of cobalt 60 on the beta carotid's in the carrot (daucus carota), using for it three different radiation dose (100, 150 and 200 kilo-rad) and analyzing them by means of the liquid chromatography technique of high resolution (HPLC)

Development of an automated chip culture system with integrated on-line monitoring for maturation culture of retinal pigment epithelial cells

Directory of Open Access Journals (Sweden)

Mee-Hae Kim

2017-10-01

Full Text Available In cell manufacturing, the establishment of a fully automated, microfluidic, cell culture system that can be used for long-term cell cultures, as well as for process optimization is highly desirable. This study reports the development of a novel chip bioreactor system that can be used for automated long-term maturation cultures of retinal pigment epithelial (RPE cells. The system consists of an incubation unit, a medium supply unit, a culture observation unit, and a control unit. In the incubation unit, the chip contains a closed culture vessel (2.5 mm diameter, working volume 9.1 μL, which can be set to 37 °C and 5% CO2, and uses a gas-permeable resin (poly- dimethylsiloxane as the vessel wall. RPE cells were seeded at 5.0 × 104 cells/cm2 and the medium was changed every day by introducing fresh medium using the medium supply unit. Culture solutions were stored either in the refrigerator or the freezer, and fresh medium was prepared before any medium change by warming to 37 °C and mixing. Automated culture was allowed to continue for 30 days to allow maturation of the RPE cells. This chip culture system allows for the long-term, bubble-free, culture of RPE cells, while also being able to observe cells in order to elucidate their cell morphology or show the presence of tight junctions. This culture system, along with an integrated on-line monitoring system, can therefore be applied to long-term cultures of RPE cells, and should contribute to process control in RPE cell manufacturing.

[Biological characteristics of mesenchymal stem cell and hematopoietic stem cell in the co-culture system].

Science.gov (United States)

Wei, Wei; Xu, Chao; Ye, Zhi-Yong; Huang, Xiao-Jun; Yuan, Jia-En; Ma, Tian-Bao; Lin, Han-Biao; Chen, Xiu-Qiong

2016-10-25

The aim of the present study was to obtain the qualified hematopoietic stem/progenitor cells (HSC/HPC) and human umbilical cord-mesenchymal stem cells (MSC) in vitro in the co-culture system. Cord blood mononuclear cells were separated from umbilical cord blood by Ficoll lymphocyte separation medium, and then CD34 + HSC was collected by MACS immunomagnetic beads. The selected CD34 + HSC/HPC and MSC were transferred into culture flask. IMDM culture medium with 15% AB-type cord plasma supplemented with interleukin-3 (IL-3), IL-6, thrombopoietin (TPO), stem cell factor (SCF) and FMS-like tyrosine kinase 3 ligand (Flt-3L) factors were used as the co-culture system for the amplification of HSC/HPC and MSC. The cellular growth status and proliferation on day 6 and 10 after co-culture were observed by using inverted microscope. The percentage of positive expression of CD34 in HSC/HPC, as well as the percentages of positive expressions of CD105, CD90, CD73, CD45, CD34 and HLA-DR in the 4 th generation MSC, was tested by flow cytometry. Semisolid colony culture was used to test the HSC/HPC colony forming ability. The osteogenic, chondrogenesis and adipogenic ability of the 4 th generation MSC were assessed. The karyotype analysis of MSC was conducted by colchicines. The results demonstrated that the HSC/HPC of co-culture group showed higher ability of amplification, CFU-GM and higher CD34 + percentage compared with the control group. The co-cultured MSC maintained the ability to differentiate into bone cells, fat cells and chondrocytes. And the karyotype stability of MSC remained normal. These results reveal that the appropriate co-culture system for MSC and HSC is developed, and via this co-culture system we could gain both two kinds of these cells. The MSCs under the co-culture system maintain the biological characteristics. The CFU-GM ability, cell counting and the flow cytometry results of HSC/HPC under the co-culture system are conform to the criterion, showing that

Production of betalaines by Myrtillocactus cell cultures. Passage from heterotrophic state to autotrophic state with Asparagus cell cultures

Energy Technology Data Exchange (ETDEWEB)

Bulard, C; Mary, J; Chaumont, D; Gudin, C

1982-11-01

Myrtillocactus tissue cultures are grown from the epicotyl of young plantlets. With an appropriate growing medium it is possible, after transfer of fragments of these cultures to a liquid environment, to obtain dissociation and proliferation of cells. The production of betalaic pigments is induced in solid surroundings by adjustement of the growing medium composition and can be maintained in a liquid environment. The multiplication of pigmented cells in suspension may thus be obtained. The conversion of Asparagus cell suspensions from the heterotrophic state (use of lactose as source of carbon) to the autotrophic state (carbon supplied by CO/sub 2/) is obtained by a gradual reduction in the sugar concentration of the medium combined with a rise in the CO/sub 2/ content of the gas mixture atmosphere injected into the cultivator. The passage to the autotrophic state of a Myrtillocactus suspension would enable the production conditions of a metabolite (Betalaine) to be studied by micro-algae culture techniques.

Hypoxic contraction of cultured pulmonary vascular smooth muscle cells

International Nuclear Information System (INIS)

Murray, T.R.; Chen, L.; Marshall, B.E.; Macarak, E.J.

1990-01-01

The cellular events involved in generating the hypoxic pulmonary vasoconstriction response are not clearly understood, in part because of the multitude of factors that alter pulmonary vascular tone. The goal of the present studies was to determine if a cell culture preparation containing vascular smooth muscle (VSM) cells could be made to contract when exposed to a hypoxic atmosphere. Cultures containing only fetal bovine pulmonary artery VSM cells were assessed for contractile responses to hypoxic stimuli by two methods. In the first, tension forces generated by cells grown on a flexible growth surface (polymerized polydimethyl siloxane) were manifested as wrinkles and distortions of the surface under the cells. Wrinkling of the surface was noted to progressively increase with time as the culture medium bathing the cells was made hypoxic (PO2 approximately 25 mmHg). The changes were sometimes reversible upon return to normoxic conditions and appeared to be enhanced in cells already exhibiting evidence of some baseline tone. Repeated passage in culture did not diminish the hypoxic response. Evidence for contractile responses to hypoxia was also obtained from measurements of myosin light chain (MLC) phosphorylation. Conversion of MLC to the phosphorylated species is an early step in the activation of smooth muscle contraction. Lowering the PO2 in the culture medium to 59 mmHg caused a 45% increase in the proportion of MLC in the phosphorylated form as determined by two-dimensional gel electrophoresis. Similarly, cultures preincubated for 4 h with 32P and then exposed to normoxia or hypoxia for a 5-min experimental period showed more than twice as much of the label in MLCs of the hypoxic cells

Contributions of 3D Cell Cultures for Cancer Research.

Science.gov (United States)

Ravi, Maddaly; Ramesh, Aarthi; Pattabhi, Aishwarya

2017-10-01

Cancer cell lines have contributed immensely in understanding the complex physiology of cancers. They are excellent material for studies as they offer homogenous samples without individual variations and can be utilised with ease and flexibility. Also, the number of assays and end-points one can study is almost limitless; with the advantage of improvising, modifying or altering several variables and methods. Literally, a new dimension to cancer research has been achieved by the advent of 3Dimensional (3D) cell culture techniques. This approach increased many folds the ways in which cancer cell lines can be utilised for understanding complex cancer biology. 3D cell culture techniques are now the preferred way of using cancer cell lines to bridge the gap between the 'absolute in vitro' and 'true in vivo'. The aspects of cancer biology that 3D cell culture systems have contributed include morphology, microenvironment, gene and protein expression, invasion/migration/metastasis, angiogenesis, tumour metabolism and drug discovery, testing chemotherapeutic agents, adaptive responses and cancer stem cells. We present here, a comprehensive review on the applications of 3D cell culture systems for these aspects of cancers. J. Cell. Physiol. 232: 2679-2697, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Evaluation of a dental pulp-derived cell sheet cultured on amniotic membrane substrate.

Science.gov (United States)

Honjo, Ken-ichi; Yamamoto, Toshiro; Adachi, Tetsuya; Amemiya, Takeshi; Mazda, Osam; Kanamura, Narisato; Kita, Masakazu

2015-01-01

Mesenchymal stem cells (MSC) are transplanted for periodontal tissue regeneration, and the periodontal ligament (PDL) is regenerated using a cultured cell sheet. This cultured cell sheet is prepared using PDL-derived cells, growth factors, and amniotic membrane (AM). Dental pulp (DP)-derived cells can be easily obtained from extracted wisdom teeth, proliferate rapidly, and are less susceptible to bacterial infection than PDL-derived cells. Thus, to prepare a novel cell sheet, DP-derived cells were cultured on AM as a culture substrate for immunohistochemical examination. Wisdom teeth extracted from three adults were cut along the cement-enamel border. DP tissue was collected, minced, and primarily cultured. After three or four passage cultures, DP-derived cells were cultured on AM, followed by hematoxylin-eosin (H-E) and immunofluorescence staining. DP-derived cells cultured on AM formed a layered structure. Cells positive for vimentin, Ki-67, ZO-1, desmoplakin, CD29, 44, 105 or 146, STRO-1, collagen IV or VII or laminin 5 or α5 chain were localized. DP-derived cells proliferated on AM, while retaining the properties of DP, which allowed the cultured cell sheet to be prepared. In addition, the cultured cell sheet contained MSC, which suggests its potential application in periodontal tissue regeneration.

Cytotoxicity of TSP in 3D Agarose Gel Cultured Cell.

Directory of Open Access Journals (Sweden)

Song-I Chun

Full Text Available A reference reagent, 3-(trimethylsilyl propionic-2, 2, 3, 3-d4 acid sodium (TSP, has been used frequently in nuclear magnetic resonance (NMR and magnetic resonance spectroscopy (MRS as an internal reference to identify cell and tissue metabolites, and determine chemical and protein structures. This reference material has been exploited for the quantitative and dynamic analyses of metabolite spectra acquired from cells. The aim of this study was to evaluate the cytotoxicity of TSP on three-dimensionally, agarose gel, cultured cells.A human osteosarcoma cell line (MG-63 was selected, and cells were three dimensionally cultured for two weeks in an agarose gel. The culture system contained a mixture of conventional culture medium and various concentrations (0, 1, 3, 5, 7, 10, 20 30 mM of TSP. A DNA quantification assay was conducted to assess cell proliferation using Quant-iT PicoGreen dsDNA reagent and kit, and cell viability was determined using a LIVE/DEAD Viability/Cytotoxicity kit. Both examinations were performed simultaneously at 1, 3, 7 and 14 days from cell seeding.In this study, the cytotoxicity of TSP in the 3D culture of MG-63 cells was evaluated by quantifying DNA (cell proliferation and cell viability. High concentrations of TSP (from 10 to 30 mM reduced both cell proliferation and viability (to 30% of the control after one week of exposure, but no such effects were found using low concentrations of TSP (0-10 mM.This study shows that low concentrations of TSP in 3D cell culture medium can be used for quantitative NMR or MRS examinations for up to two weeks post exposure.

Establishment and characterization of American elm cell suspension cultures

Science.gov (United States)

Steven M. Eshita; Joseph C. Kamalay; Vicki M. Gingas; Daniel A. Yaussy

2000-01-01

Cell suspension cultures of Dutch elm disease (DED)-tolerant and DED-susceptible American elms clones have been established and characterized as prerequisites for contrasts of cellular responses to pathogen-derived elicitors. Characteristics of cultured elm cell growth were monitored by A700 and media conductivity. Combined cell growth data for all experiments within a...

Epithelial Cell Cultures

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Imran S. Chaudhry

2011-01-01

Full Text Available The biological effects of only a finite number of tobacco toxins have been studied. Here, we describe exposure of cultures of human bronchial epithelial cells to low concentrations of tobacco carcinogens: nickel sulphate, benzo(bfluoranthene, N-nitrosodiethylamine, and 4-(methylnitrosamino-1-(3-pyridyl-1-butanone (NNK. After a 24-hour exposure, EGFR was expressed in cell membrane and cytoplasm, BCL-2 was expressed only in the irregular nuclei of large atypical cells, MKI67 was expressed in nuclei with no staining in larger cells, cytoplasmic BIRC5 with stronger nuclear staining was seen in large atypical cells, and nuclear TP53 was strongly expressed in all cells. After only a 24-hour exposure, cells exhibited atypical nuclear and cytoplasmic features. After a 48-hour exposure, EGFR staining was localized to the nucleus, BCL-2 was slightly decreased in intensity, BIRC5 was localized to the cytoplasm, and TP53 staining was increased in small and large cells. BCL2L1 was expressed in both the cytoplasm and nuclei of cells at 24- and 48-hour exposures. We illustrate that short-termexposure of a bronchial epithelial cell line to smoking-equivalent concentrations of tobacco carcinogens alters the expression of key proliferation regulatory genes, EGFR, BCL-2, BCL2L1, BIRC5, TP53, and MKI67, similar to that reported in biopsy specimens of pulmonary epithelium described to be preneoplastic lesions.

Stimulation and support of haemopoietic stem cell proliferation by irradiated stroma cell colonies in bone marrow cell culture in vitro

International Nuclear Information System (INIS)

Mori, K.J.; Izumi, Hiroko; Seto, Akira

1981-01-01

A culture system was established in which haemopoietic stem cells can undergo a recovery proliferation after a depletion of the stem cells, completely in vitro. To elucidate the source of the stimulatory factors, normal bone marrow cells were overlayed on top of the irradiated adherent 'stromal' cell colonies in the bone marrow cell culture. This stimulated the proliferation of haemopoietic stem cells in the cultured cells in suspension. The present results indicate that the stromal cells produce factors which stimulate stem cell proliferation. Whether the stimulation is evoked by direct cell-cell interactions or by humoral factors is as yet to be studied. (author)

Immunocytochemical characterization of primary cell culture in canine transmissible venereal tumor

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Luis M.M. Flórez

Full Text Available Abstract: Immunochemistry with anti-vimentin, anti-lysozyme, anti-alpha 1 antitrypsin, anti-CD3 and anti-CD79α antibodies has been used for characterization of primary cell culture in the transmissible venereal tumor (TVT. Samples for primary cell culture and immunohistochemistry assays were taken from eight dogs with cytological and clinical diagnosis of TVT. To validate the immunochemical results in the primary cell culture of TVT, a chromosome count was performed. For the statistical analysis, the Mann-Whitney test with p<0.05 was used. TVT tissues and culture cells showed intense anti-vimentin immunoreactivity, lightly to moderate immunoreactivity for anti-lysozyme, and mild for anti-alpha-antitrypsin. No marking was achieved for CD3 and CD79α. All culture cells showed chromosomes variable number of 56 to 68. This is the first report on the use of immunocytochemical characterization in cell culture of TVT. Significant statistic difference between immunochemistry in tissue and culture cell was not established, what suggests that the use of this technique may provide greater certainty for the confirmation of tumors in the primary culture. This fact is particularly important because in vitro culture of tumor tissues has been increasingly used to provide quick access to drug efficacy and presents relevant information to identify potential response to anticancer medicine; so it is possible to understand the behavior of the tumor.

The Effect of Isabgol (Plantago psyllium Mucilage and Shiraz Thyme Essential Oils on Microbial Load and Improving Shelf Life of Fresh-Cut Carrot

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M. Azizi

2016-02-01

Full Text Available Introduction: Fresh-cut produce graduated to retail during the1990s, especially for lettuce, cabbage, carrots and other similar vegetables. The high microbial loads of these products after harvest can be substantially reduced through a cleaning in flowing chlorinated water and adistribution under ensured controlledrefrigeration. Therefore, a good number of convenient ready-to-use greens were launched to the market in the past decade. Nowadays, theuse of this technology to achieve similar results in fruit products is one of the most challengingtargets for processors. However, there is anumber of issues that still need to beovercomebeforefresh-cut fruit commodities can be sparked off to anoutstanding position in the segment of lightly-treatedrefrigerated foods. The importance of freshly cut products increases day by day. Tissue and cell rupture leads to a decrease in the shelf life of these products. On the other hand, these products due to increased enzyme activity, respiration rate and microbiological considerations that affect the health of these productsrequires highly attention.To increase the shelf life of the products and prevent undesirable changes in cut slices of fruit or vegetables a coating on the surface of these products has been suggested. Mucilages and essential oils of herbs are natural compounds that can be used to create such covers. The advantages of these coatings are their bactericidal effect, maintenanceof pleasant taste and other physical and chemical characteristics of the product and even decrease of environmental pollution. In this research, the effect of natural compounds such as Zataria multiflora essential oil (EO and Plantagopsyllium mucilage on storage life and microbial load of fresh cut carrot was studied. Materials and Methods: The research was conducted in two separate experiments on fresh-cut carrot: In the first experiment, the effect of different concentrations of Plantago psyllium mucilage (0,100, 200, and

Irradiated murine fibroblasts as feeder layer used in human cell culture

International Nuclear Information System (INIS)

Almeida, Tiago L.; Klingbeil, Fatima G.; Yoshito, Daniele; Caproni, Priscila; Mathor, Monica B.; Herson, Marisa R.

2007-01-01

In 1975, Rheinwald and Green published an in vitro model for keratinocyte cell cultures in which the use of murine fibroblasts, as a feeder layer was introduced. These cells are modified fibroblasts, which presence render keratinocyte cells to remain proliferative for longer periods of time. This optimization of culture outputs has allowed for several clinical applications of confluent keratinocyte cultures as skin substitutes or wound dressings in situations such as post burn extensive skin loss, loss of oral mucosa, and other skin disorders. Nevertheless, proliferation of fibroblast in co-culture with keratinocytes must be controlled by anti-proliferative measures such as irradiation; at the same time, keratinocytes require specific nutrients in the culture medium, which may interfere with the fibroblast feeder layer viability. Therefore, the thorough understanding of the impact of different issues such as culture media composition, irradiation dose and pre-plating storage conditions of irradiated fibroblast to be used as feeder layer in these co-culture systems is important. In this work, changes as far as viability and proliferative rates of irradiated fibroblasts in culture were evaluated in relation to the type of culture medium used, dose of gamma radiation exposure, storage and timing of cell plating post irradiation. Results indicate that the type of culture medium used and time-lag between irradiation, refrigeration and plating of irradiated cells do not have significant impact in culture outcomes. However, the dose of gamma radiation administered to the cells may influence the final quality of these cells if to be used as a feeder layer. (author)

Animal-cell culture media: History, characteristics, and current issues.

Science.gov (United States)

Yao, Tatsuma; Asayama, Yuta

2017-04-01

Cell culture technology has spread prolifically within a century, a variety of culture media has been designed. This review goes through the history, characteristics and current issues of animal-cell culture media. A literature search was performed on PubMed and Google Scholar between 1880 and May 2016 using appropriate keywords. At the dawn of cell culture technology, the major components of media were naturally derived products such as serum. The field then gradually shifted to the use of chemical-based synthetic media because naturally derived ingredients have their disadvantages such as large batch-to-batch variation. Today, industrially important cells can be cultured in synthetic media. Nevertheless, the combinations and concentrations of the components in these media remain to be optimized. In addition, serum-containing media are still in general use in the field of basic research. In the fields of assisted reproductive technologies and regenerative medicine, some of the medium components are naturally derived in nearly all instances. Further improvements of culture media are desirable, which will certainly contribute to a reduction in the experimental variation, enhance productivity among biopharmaceuticals, improve treatment outcomes of assisted reproductive technologies, and facilitate implementation and popularization of regenerative medicine.

The study of the deuterium isotopic fractionation through the cell membrane of the plant

International Nuclear Information System (INIS)

Berdea, P.; Cuna, Stela; Deliu, C.

2002-01-01

The purpose of this study is to prove that there is a water deuterium isotope fractionation when the water passes through the cell membrane. The carrots (Daucus carota) were grown in vitro in a Murashige and Skoog mineral-salt medium and have been exposed to a water solution with a uniform isotopic content. After seven days the cell culture was filtered and the cell water was vacuum extracted. The water from aqueous solution and the cell water were analyzed for hydrogen by isotope ratio mass spectrometry. The procedure was repeated for 14 and 21 day old cell cultures. The measurements have revealed a water deuterium isotopic fractionation between extra-cellular water and cellular water. The deuterium content was found to be higher within the cells by 10 o / oo for non-embryonic cells and 13 o / oo for the embryonic cells. This fractionation is a non-evaporative fractionation between intracellular and extra-cellular water and it represents a new step in the overall fractionation of deuterium water in the plants. The existence of such isotopic fractionation through the cell membrane implies that the relationship between the deuterium content of cellulose nitrate in plant and meteoric water should be revised. Also, this finding is of interest for understanding the balance and dynamics of the hydrogen isotopes in the environment. (authors)

Cell Culture Assay for Human Noroviruses [response

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Straub, Tim M.; Honer Zu Bentrup, Kerstin; Orosz Coghlan, Patricia; Dohnalkova, Alice; Mayer, Brooke K.; Bartholomew, Rachel A.; Valdez, Catherine O.; Bruckner-Lea, Cindy J.; Gerba, Charles P.; Abbaszadegan, Morteza A.; Nickerson, Cheryl A.

2007-07-01

We appreciate the comments provided by Leung et al., in response to our recently published article “In Vitro Cell Culture Infectivity Assay for Human Noroviruses” by Straub et al. (1). The specific aim of our project was to develop an in vitro cell culture infectivity assay for human noroviruses (hNoV) to enhance risk assessments when they are detected in water supplies. Reverse transcription (RT) qualitative or quantitative PCR are the primary assays for waterborne NoV monitoring. However, these assays cannot distinguish between infectious vs. non-infectious virions. When hNoV is detected in water supplies, information provided by our infectivity assay will significantly improve risk assessment models and protect human health, regardless of whether we are propagating NoV. Indeed, in vitro cell culture infectivity assays for the waterborne pathogen Cryptosporidium parvum that supplement approved fluorescent microscopy assays, do not result in amplification of the environmentally resistant hard-walled oocysts (2). However, identification of life cycle stages in cell culture provides evidence of infectious oocysts in a water supply. Nonetheless, Leung et al.’s assertion regarding the suitability of our method for the in vitro propagation of high titers of NoV is valid for the medical research community. In this case, well-characterized challenge pools of virus would be useful for developing and testing diagnostics, therapeutics, and vaccines. As further validation of our published findings, we have now optimized RT quantitative PCR to assess the level of viral production in cell culture, where we are indeed finding significant increases in viral titer. The magnitude and time course of these increases is dependent on both virus strain and multiplicity of infection. We are currently preparing a manuscript that will discuss these findings in greater detail, and the implications this may have for creating viral challenge pools

Rheological characteristics of cell suspension and cell culture of Perilla frutescens.

Science.gov (United States)

Zhong, J J; Seki, T; Kinoshita, S; Yoshida, T

1992-12-05

Physical properties such as viscosity, fluid dynamic behavior of cell suspension, and size distribution of cell aggregates of a plant, Perilla frustescens, cultured in a liquid medium were studied. As a result of investigations using cells harvester after 12 days of cultivation in a flask, it was found that the apparent viscosity of the cell suspension did not change with any variation of cell concentration below 5 g dry cell/L but markedly increased when the cell concentration increased over 12.8 g dry cell/L. The cell suspension exhibited the characteristics of a Bingham plastic fluid with a small yield stress. The size of cell aggregates in the range 74 to 500 mum did not influence the rheological characteristics of the cell suspension. The rheological characteristics of cultivation mixtures of P. frutescens cultivated in a flask and in a bioreactor were also investigated. The results showed that the flow characteristics of the cell culture could be described by a Bingham plastic model. At the later stage of cultivation, the apparent viscosity increased steadily, even though the biomass concentration (by dry weight) decreased, due to the increase of individual cell size. (c) 1992 John Wiley & Sons, Inc.

Influence of thermal processing on hydrolysis and stability of folate poly-gamma-glutamates in broccoli (Brassica oleracea var. italica), carrot (Daucus carota) and tomato (Lycopersicon esculentum).

Science.gov (United States)

Munyaka, Ann Wambui; Verlinde, Philippe; Mukisa, Ivan Muzira; Oey, Indrawati; Van Loey, Ann; Hendrickx, Marc

2010-04-14

The folate poly-gamma-glutamate profile, their concentrations, and hydrolysis by endogenous gamma-glutamyl hydrolase (GGH) were evaluated in broccoli, carrot and tomato. Further studies on the effect of time and temperature on folate poly-gamma-glutamate hydrolysis and stability were carried out in broccoli since this vegetable showed the highest long-chain and total folate poly-gamma-glutamate concentration. The evolution of l-ascorbic acid, total phenols and Trolox equivalent antioxidant capacity (TEAC) values was evaluated in parallel. Upon thermal inactivation of GGH prior to crushing, it was observed that broccoli, carrot and tomato contained poly-gamma-glutamates with one to seven glutamate residues but differed in the predominant poly-gamma-glutamates. Crushing of raw broccoli, carrot and tomato resulted in significant poly-gamma-glutamate profile changes in broccoli and carrot (indicating GGH-catalyzed hydrolysis) but not in tomato. In this study, the actual crushing of raw broccoli matrix had a greater effect on folate poly-gamma-glutamate hydrolysis than incubation conditions (0-30 min at 25-55 degrees C). During treatments at 25-140 degrees C, folate retention was higher at 80 and 100 degrees C than at the other temperatures. A similar trend in thermal stability was observed for folates, vitamin C, total phenols and TEAC value, an indication that conditions that result in endogenous antioxidants degradation might also result in folate degradation.

Co-culture with Sertoli cells promotes proliferation and migration of umbilical cord mesenchymal stem cells

International Nuclear Information System (INIS)

Zhang, Fenxi; Hong, Yan; Liang, Wenmei; Ren, Tongming; Jing, Suhua; Lin, Juntang

2012-01-01

Highlights: ► Co-culture of Sertoli cells (SCs) with human umbilical cord mesenchymal stem cells (UCMSCs). ► Presence of SCs dramatically increased proliferation and migration of UCMSCs. ► Presence of SCs stimulated expression of Mdm2, Akt, CDC2, Cyclin D, CXCR4, MAPKs. -- Abstract: Human umbilical cord mesenchymal stem cells (hUCMSCs) have been recently used in transplant therapy. The proliferation and migration of MSCs are the determinants of the efficiency of MSC transplant therapy. Sertoli cells are a kind of “nurse” cells that support the development of sperm cells. Recent studies show that Sertoli cells promote proliferation of endothelial cells and neural stem cells in co-culture. We hypothesized that co-culture of UCMSCs with Sertoli cells may also promote proliferation and migration of UCMSCs. To examine this hypothesis, we isolated UCMSCs from human cords and Sertoli cells from mouse testes, and co-cultured them using a Transwell system. We found that UCMSCs exhibited strong proliferation ability and potential to differentiate to other cell lineages such as osteocytes and adipocytes. The presence of Sertoli cells in co-culture significantly enhanced the proliferation and migration potential of UCMSCs (P < 0.01). Moreover, these phenotypic changes were accompanied with upregulation of multiple genes involved in cell proliferation and migration including phospho-Akt, Mdm2, phospho-CDC2, Cyclin D1, Cyclin D3 as well as CXCR4, phospho-p44 MAPK and phospho-p38 MAPK. These findings indicate that Sertoli cells boost UCMSC proliferation and migration potential.

Endothelial cells stimulate growth of normal and cancerous breast epithelial cells in 3D culture

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Magnusson Magnus K

2010-07-01

Full Text Available Abstract Background Epithelial-stromal interaction provides regulatory signals that maintain correct histoarchitecture and homeostasis in the normal breast and facilitates tumor progression in breast cancer. However, research on the regulatory role of the endothelial component in the normal and malignant breast gland has largely been neglected. The aim of the study was to investigate the effects of endothelial cells on growth and differentiation of human breast epithelial cells in a three-dimensional (3D co-culture assay. Methods Breast luminal and myoepithelial cells and endothelial cells were isolated from reduction mammoplasties. Primary cells and established normal and malignant breast cell lines were embedded in reconstituted basement membrane in direct co-culture with endothelial cells and by separation of Transwell filters. Morphogenic and phenotypic profiles of co-cultures was evaluated by phase contrast microscopy, immunostaining and confocal microscopy. Results In co-culture, endothelial cells stimulate proliferation of both luminal- and myoepithelial cells. Furthermore, endothelial cells induce a subpopulation of luminal epithelial cells to form large acini/ducts with a large and clear lumen. Endothelial cells also stimulate growth and cloning efficiency of normal and malignant breast epithelial cell lines. Transwell and gradient co-culture studies show that endothelial derived effects are mediated - at least partially - by soluble factors. Conclusion Breast endothelial cells - beside their role in transporting nutrients and oxygen to tissues - are vital component of the epithelial microenvironment in the breast and provide proliferative signals to the normal and malignant breast epithelium. These growth promoting effects of endothelial cells should be taken into consideration in breast cancer biology.

Co-culture with Sertoli cells promotes proliferation and migration of umbilical cord mesenchymal stem cells

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Zhang, Fenxi, E-mail: fxzhang0824@gmail.com [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Hong, Yan; Liang, Wenmei [Department of Histology and Embryology, Guiyang Medical University, Guizhou 550004, People' s Republic of China (China); Ren, Tongming [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Jing, Suhua [ICU Center, The Third Hospital of Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Lin, Juntang [Stem Cell Center, Xinxiang Medical University, Henan 453003, People' s Republic of China (China)

2012-10-12

Highlights: Black-Right-Pointing-Pointer Co-culture of Sertoli cells (SCs) with human umbilical cord mesenchymal stem cells (UCMSCs). Black-Right-Pointing-Pointer Presence of SCs dramatically increased proliferation and migration of UCMSCs. Black-Right-Pointing-Pointer Presence of SCs stimulated expression of Mdm2, Akt, CDC2, Cyclin D, CXCR4, MAPKs. -- Abstract: Human umbilical cord mesenchymal stem cells (hUCMSCs) have been recently used in transplant therapy. The proliferation and migration of MSCs are the determinants of the efficiency of MSC transplant therapy. Sertoli cells are a kind of 'nurse' cells that support the development of sperm cells. Recent studies show that Sertoli cells promote proliferation of endothelial cells and neural stem cells in co-culture. We hypothesized that co-culture of UCMSCs with Sertoli cells may also promote proliferation and migration of UCMSCs. To examine this hypothesis, we isolated UCMSCs from human cords and Sertoli cells from mouse testes, and co-cultured them using a Transwell system. We found that UCMSCs exhibited strong proliferation ability and potential to differentiate to other cell lineages such as osteocytes and adipocytes. The presence of Sertoli cells in co-culture significantly enhanced the proliferation and migration potential of UCMSCs (P < 0.01). Moreover, these phenotypic changes were accompanied with upregulation of multiple genes involved in cell proliferation and migration including phospho-Akt, Mdm2, phospho-CDC2, Cyclin D1, Cyclin D3 as well as CXCR4, phospho-p44 MAPK and phospho-p38 MAPK. These findings indicate that Sertoli cells boost UCMSC proliferation and migration potential.

Development of Cell Culture Microdevice Actuated by Piezoelectric Thin Films for Delivering Mechanical Vibratory Stimuli to Cells

International Nuclear Information System (INIS)

Yamada, Y; Umegaki, G; Kawashima, T; Nagai, M; Shibata, T; Masuzawa, T; Kimura, T; Kishida, A

2012-01-01

In order to realize a cell culture microdevice actuated by piezoelectric thin films for on-chip regulation of cell functions, this paper reported on a feasibility study by using the microdevice with KOH-etched cavities surrounded by four (111) sidewalls as microchambers in order to introduce cells to be cultured. As a result, the vibration characteristic of the PZT actuator was improved by using an electric field -150 kV/cm at 70 C for 30 min in poling process. A feasibility study on cell culture for delivering mechanical vibratory stimuli to cells revealed the microdevice could be applicable to the culture with actual biological cells. In addition, it was found that O 2 -plasma treated parylene-C process could be applicable for obtaining homogeneous surface of cell culture microdevice.