Nest-mediated seed dispersal

Science.gov (United States)

Robert J. Warren; Jason P. Love; Mark A. Bradford

2017-01-01

Many plant seeds travel on the wind and through animal ingestion or adhesion; however, an overlooked dispersal mode may lurk within those dispersal modes. Viable seeds may remain attached or embedded within materials birds gather for nest building. Our objective was to determine if birds inadvertently transport seeds when they forage for plant materials to...

7 CFR 201.57a - Dormant seeds.

Science.gov (United States)

2010-01-01

... REGULATIONS Germination Tests in the Administration of the Act § 201.57a Dormant seeds. Dormant seeds are viable seeds, other than hard seeds, which fail to germinate when provided the specified germination..., or application of germination promoting chemicals. (b) The percentage of dormant seed, if present...

Enhanced seed viability and lipid compositional changes during natural ageing by suppressing phospholipase Dα in soybean seed.

Science.gov (United States)

Lee, Junghoon; Welti, Ruth; Roth, Mary; Schapaugh, William T; Li, Jiarui; Trick, Harold N

2012-02-01

Changes in phospholipid composition and consequent loss of membrane integrity are correlated with loss of seed viability. Furthermore, phospholipid compositional changes affect the composition of the triacylglycerols (TAG), i.e. the storage lipids. Phospholipase D (PLD) catalyses the hydrolysis of phospholipids to phosphatidic acid, and PLDα is an abundant PLD isoform. Although wild-type (WT) seeds stored for 33 months were non-viable, 30%-50% of PLDα-knockdown (PLD-KD) soybean seeds stored for 33 months germinated. WT and PLD-KD seeds increased in lysophospholipid levels and in TAG fatty acid unsaturation during ageing, but the levels of lysophospholipids increased more in WT than in PLD-KD seeds. The loss of viability of WT seeds was correlated with alterations in ultrastructure, including detachment of the plasma membrane from the cell wall complex and disorganization of oil bodies. The data demonstrate that, during natural ageing, PLDα affects the soybean phospholipid profile and the TAG profile. Suppression of PLD activity in soybean seed has potential for improving seed quality during long-term storage. © 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

Mobilization of Viable Tumor Cells Into the Circulation During Radiation Therapy

International Nuclear Information System (INIS)

Martin, Olga A.; Anderson, Robin L.; Russell, Prudence A.; Ashley Cox, R.; Ivashkevich, Alesia; Swierczak, Agnieszka; Doherty, Judy P.; Jacobs, Daphne H.M.; Smith, Jai; Siva, Shankar; Daly, Patricia E.; Ball, David L.

2014-01-01

Purpose: To determine whether radiation therapy (RT) could mobilize viable tumor cells into the circulation of non-small cell lung cancer (NSCLC) patients. Methods and Materials: We enumerated circulating tumor cells (CTCs) by fluorescence microscopy of blood samples immunostained with conventional CTC markers. We measured their DNA damage levels using γ-H2AX, a biomarker for radiation-induced DNA double-strand breaks, either by fluorescence-activated cell sorting or by immunofluorescence microscopy. Results: Twenty-seven RT-treated NSCLC patients had blood samples analyzed by 1 or more methods. We identified increased CTC numbers after commencement of RT in 7 of 9 patients treated with palliative RT, and in 4 of 8 patients treated with curative-intent RT. Circulating tumor cells were also identified, singly and in clumps in large numbers, during RT by cytopathologic examination (in all 5 cases studied). Elevated γ-H2AX signal in post-RT blood samples signified the presence of CTCs derived from irradiated tumors. Blood taken after the commencement of RT contained tumor cells that proliferated extensively in vitro (in all 6 cases studied). Circulating tumor cells formed γ-H2AX foci in response to ex vivo irradiation, providing further evidence of their viability. Conclusions: Our findings provide a rationale for the development of strategies to reduce the concentration of viable CTCs by modulating RT fractionation or by coadministering systemic therapies

Mobilization of Viable Tumor Cells Into the Circulation During Radiation Therapy

Energy Technology Data Exchange (ETDEWEB)

Martin, Olga A. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Molecular Radiation Biology Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); The Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC (Australia); Anderson, Robin L. [The Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC (Australia); Metastasis Research Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Russell, Prudence A. [Department of Anatomical Pathology, St. Vincent Hospital, Fitzroy, VIC (Australia); Ashley Cox, R. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Ivashkevich, Alesia [Molecular Radiation Biology Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Laboratory of DNA Repair and Genomics, Centre for Innate Immunity and Infectious Disease, Monash Institute for Medical Research, Monash University, Clayton, VIC (Australia); Swierczak, Agnieszka; Doherty, Judy P. [Metastasis Research Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Jacobs, Daphne H.M. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Smith, Jai [Molecular Radiation Biology Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Siva, Shankar; Daly, Patricia E. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Ball, David L. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); The Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC (Australia); and others

2014-02-01

Purpose: To determine whether radiation therapy (RT) could mobilize viable tumor cells into the circulation of non-small cell lung cancer (NSCLC) patients. Methods and Materials: We enumerated circulating tumor cells (CTCs) by fluorescence microscopy of blood samples immunostained with conventional CTC markers. We measured their DNA damage levels using γ-H2AX, a biomarker for radiation-induced DNA double-strand breaks, either by fluorescence-activated cell sorting or by immunofluorescence microscopy. Results: Twenty-seven RT-treated NSCLC patients had blood samples analyzed by 1 or more methods. We identified increased CTC numbers after commencement of RT in 7 of 9 patients treated with palliative RT, and in 4 of 8 patients treated with curative-intent RT. Circulating tumor cells were also identified, singly and in clumps in large numbers, during RT by cytopathologic examination (in all 5 cases studied). Elevated γ-H2AX signal in post-RT blood samples signified the presence of CTCs derived from irradiated tumors. Blood taken after the commencement of RT contained tumor cells that proliferated extensively in vitro (in all 6 cases studied). Circulating tumor cells formed γ-H2AX foci in response to ex vivo irradiation, providing further evidence of their viability. Conclusions: Our findings provide a rationale for the development of strategies to reduce the concentration of viable CTCs by modulating RT fractionation or by coadministering systemic therapies.

Improving tomato seed quality- challenges and possibilities

DEFF Research Database (Denmark)

Shrestha, Santosh

The thesis investigates the possibility of using single seed near-infrared (NIR) spectroscopy, multispectral imaging (MSI) and NIR hyperspectral imaging (NIR-HSI) in combination with chemometrics for rapid determination of the tomato seed quality. The results of the PhD study are compiled in four...... manuscripts (MS). These non-destructive methods show the potential of sorting tomato seeds as per their viability and varietal identity. The results are discussed in the context of possible contribution from these methods in the improvement of the seed quality in Nepal. In MS I, potential application of NIR...... spectroscopy in combination with chemometrics for prediction of tomato seed viability is demonstrated. The work in MS I also emphasises on identifying the important NIR spectral regions for the chemometric model that are relevant to the separation of viable and non-viable seeds. The NIR-HIS method was also...

UVA-induced ROS generation inhibition by Oenothera paradoxa defatted seeds extract and subsequent cell death in human dermal fibroblasts.

Science.gov (United States)

Jaszewska, Edyta; Soin, Magdalena; Filipek, Agnieszka; Naruszewicz, Marek

2013-09-05

UVA radiation stimulates the production of reactive oxygen species (ROS), which react with lipids, proteins and other intracellular molecules leading to oxidative stress, cellular damage and ultimately cell death. There is, therefore, a growing need for substances exhibiting antioxidant activity, which may support repair mechanisms of the skin. This study evaluates the protective effect of the aqueous Oenothera paradoxa Hudziok defatted seeds extract, rich in polyphenolic compounds, against UVA (25 and 50J/cm(2))-induced changes in normal human dermal fibroblasts (NHDFs). The tested extract (0.1-10μg/ml) has decreased, in a concentration-dependent fashion, the UVA-induced release of lactate dehydrogenase (LDH) into the culture medium, the ROS production (with the use of 2',7'-dichlorodihydrofluorescein diacetate) and lipid peroxidation (utilizing redox reactions with ferrous ions) as compared to the control cells (incubated without the extract). Moreover, the extract increased the number of viable (calcein positive) cells decreasing the number of cells in late apoptosis (annexin V-FITC and propidium iodide positive). Thus our results show that O. paradoxa defatted seeds extract may be beneficial for the prevention of UVA skin damage. Copyright © 2013 Elsevier B.V. All rights reserved.

Weed seed spread and its prevention: The role of roadside wash down.

Science.gov (United States)

Bajwa, Ali Ahsan; Nguyen, Thi; Navie, Sheldon; O'Donnell, Chris; Adkins, Steve

2018-02-15

Vehicles are one of the major vectors of long-distance weed seed spread. Viable seed removed from vehicles at roadside wash down facilities was studied at five locations in central Queensland, Australia over a 3-year period. Seed from 145 plant species, belonging to 34 different families, were identified in the sludge samples obtained from the wet particulate matter collection pit of the wash down facilities. Most of the species were annual forbs (50%) with small or very small seed size (weed was observed in these samples. More parthenium weed seed were found in the Rolleston facility and in the spring, but its seed was present in all facilities and in all seasons. The average number of viable seed found within every ton of dry particulate matter removed from vehicles was ca. 68,000. Thus, a typical wash down facility was removing up to ca. 335,000 viable seed from vehicles per week, of which ca. 6700 were parthenium weed seed. Furthermore, 61% of these seed (ca. 200,000) were from introduced species, and about half of these (35% of total) were from species considered to be weeds. Therefore, the roadside wash down facilities found throughout Queensland can remove a substantial amount of viable weed seed from vehicles, including the invasive parthenium weed, and the use of such facilities should be strongly encouraged. Copyright © 2017 Elsevier Ltd. All rights reserved.

Use of multispectral images and chemometrics in tomato seed studies

DEFF Research Database (Denmark)

Shrestha, Santosh; Deleuran, Lise Christina; Gislum, René

During the production of tomato seeds, green tomatoes are normally discarded before seed extraction irrespective of their maturity stage. Studies indicate that seeds from green tomatoes may reach be able to reach full germination capacity. Thus the potential of multispectral imaging for non......-destructive discrimination of seeds based on their germination capacity was investigated. A total of 840 seeds extracted from green and red tomatoes were divided into two sets; a training set and a test set consisting of 648 and 192 seeds respectively. Each set consisted of 96 seeds from green tomatoes. The multispectral...... images of the seeds were captured and normalized canonical discriminant analysis was used to analyse the images. Germination tests were performed and seeds that subsequently germinated were recorded as viable. The viable seeds were classified with 99% and 98% accuracy for the training and test set...

Inheritance of fresh seed dormancy in groundnut

African Journals Online (AJOL)

SERVER

2008-02-19

Feb 19, 2008 ... Seed dormancy has been defined as the failure of an intact, viable seed to ... expected ratio of 3 dormant : 1 non-dormant seeds in F2 generations of crosses of ICGV 87378 and ICGV ... ICGV 87378 x ICGV 86158. 163. 0. 163.

Characterisation of Castor (Ricinus communis L. Seed Quality Using Fourier Transform Near-Infrared Spectroscopy in Combination with Multivariate Data Analysis

Directory of Open Access Journals (Sweden)

René Gislum

2018-04-01

Full Text Available The potential of single-seed near-infrared (NIR spectroscopy was investigated to characterise castor seeds based on their seed viability and seed oil content. Distinct differences between viable and non-viable seeds were observed in the principal component analysis (PCA analysis. Furthermore, the PCA compared heavy and medium seeds with light seeds, which were comparable to the clusters of viable and non-viable seeds, respectively. Prediction accuracies of 98.7% and 99.6% were obtained with the partial least squares discriminant analysis (PLS-DA model with a classification error rate of 0.8% and 1.1% for the training set and test set, respectively. The NIR spectral regions having chemical information from the oil in castor seeds were found to be vital for determination of seed viability.

Modelling the number of viable vegetative cells of Bacillus cereus passing through the stomach

NARCIS (Netherlands)

Wijnands, L.M.; Pielaat, A.; Dufrenne, J.B.; Zwietering, M.H.; Leusden, van F.M.

2009-01-01

Aims: Model the number of viable vegetative cells of B. cereus surviving the gastric passage after experiments in simulated gastric conditions. Materials and Methods: The inactivation of stationary and exponential phase vegetative cells of twelve different strains of Bacillus cereus, both mesophilic

Selection of viable cell subpopulations from murine tumours using FACS

International Nuclear Information System (INIS)

Chaplin, D.J.; Durand, R.E.; Olive, P.L.

1985-01-01

The authors developed a technique which enables isolation of viable tumour cells subpopulation as a function of their distance from the blood supply. The basis for this separation procedure is that the fluorochrome, Hoechst 33342, as a result of its high avidity for cellular DNA, exhibits a marked diffusion/consumption gradient when it has to pass through several cell layers. As a result intravenous injection of Hoechst 33342 into tumour bearing animals, results in a heterogeneous straining pattern within the tumour with cells close to blood vessels being brightly fluorescent while those more distant are less intensely stained. Since these differences in staining intensity persist after tumour disaggregation, cells can be sorted into subpopulations on the basis of their fluorescence intensity using a fluorescence activated cell sorter. This technique offers the unique possibility of identifying the location of those cell subpopulations resistant to treatment with either radiation or chemotherapeutic drugs

Study on Development of Non-Destructive Measurement Technique for Viability of Lettuce Seed (Lactuca sativa L) Using Hyperspectral Reflectance Imaging

Energy Technology Data Exchange (ETDEWEB)

Ahn, Chi Kook; Cho, Byoung Kwan [College of Agriculture and Life Science, Chungnam National University, Daejeon (Korea, Republic of); Mo, Chang Yeon [National Acadamy of Agricultural Science, Daejeon (Korea, Republic of); Kim, Moon S. [Environmental Microbial and Food Safety Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, United States Department of Agriculture, Washington (United States)

2012-10-15

In this study, the feasibility of hyperspectral reflectance imaging technique was investigated for the discrimination of viable and non-viable lettuce seeds. The spectral data of hyperspectral reflectance images with the spectral range between 750 nm and 1000 nm were used to develop PLS-DA model for the classification of viable and non-viable lettuce seeds. The discrimination accuracy of the calibration set was 81.6% and that of the test set was 81.2%. The image analysis method was developed to construct the discriminant images of non-viable seeds with the developed PLS-DA model. The discrimination accuracy obtained from the resultant image were 91%, which showed the feasibility of hyperspectral reflectance imaging technique for the mass discrimination of non-viable lettuce seeds from viable ones.

Study on Development of Non-Destructive Measurement Technique for Viability of Lettuce Seed (Lactuca sativa L) Using Hyperspectral Reflectance Imaging

International Nuclear Information System (INIS)

Ahn, Chi Kook; Cho, Byoung Kwan; Mo, Chang Yeon; Kim, Moon S.

2012-01-01

In this study, the feasibility of hyperspectral reflectance imaging technique was investigated for the discrimination of viable and non-viable lettuce seeds. The spectral data of hyperspectral reflectance images with the spectral range between 750 nm and 1000 nm were used to develop PLS-DA model for the classification of viable and non-viable lettuce seeds. The discrimination accuracy of the calibration set was 81.6% and that of the test set was 81.2%. The image analysis method was developed to construct the discriminant images of non-viable seeds with the developed PLS-DA model. The discrimination accuracy obtained from the resultant image were 91%, which showed the feasibility of hyperspectral reflectance imaging technique for the mass discrimination of non-viable lettuce seeds from viable ones.

Robotic seeding

DEFF Research Database (Denmark)

Pedersen, Søren Marcus; Fountas, Spyros; Sørensen, Claus Aage Grøn

2017-01-01

Agricultural robotics has received attention for approximately 20 years, but today there are only a few examples of the application of robots in agricultural practice. The lack of uptake may be (at least partly) because in many cases there is either no compelling economic benefit......, or there is a benefit but it is not recognized. The aim of this chapter is to quantify the economic benefits from the application of agricultural robots under a specific condition where such a benefit is assumed to exist, namely the case of early seeding and re-seeding in sugar beet. With some predefined assumptions...... with regard to speed, capacity and seed mapping, we found that among these two technical systems both early seeding with a small robot and re-seeding using a robot for a smaller part of the field appear to be financially viable solutions in sugar beet production....

Seed propagation of Allanblackia floribunda by mechanical pre ...

African Journals Online (AJOL)

Mechanical pre-germination treatments were given to the viable seeds: total removal of seed coat, sand-paper scarification and laceration. These were sown in forest topsoil and washed river sand media respectively in completely Randomized Design in the Nursery. Seeds with testa totally removed and sown in forest ...

Epitaxially grown polycrystalline silicon thin-film solar cells on solid-phase crystallised seed layers

Energy Technology Data Exchange (ETDEWEB)

Li, Wei, E-mail: weili.unsw@gmail.com; Varlamov, Sergey; Xue, Chaowei

2014-09-30

Highlights: • Crystallisation kinetic is used to analyse seed layer surface cleanliness. • Simplified RCA cleaning for the seed layer can shorten the epitaxy annealing duration. • RTA for the seed layer can improve the quality for both seed layer and epi-layer. • Epitaxial poly-Si solar cell performance is improved by RTA treated seed layer. - Abstract: This paper presents the fabrication of poly-Si thin film solar cells on glass substrates using seed layer approach. The solid-phase crystallised P-doped seed layer is not only used as the crystalline template for the epitaxial growth but also as the emitter for the solar cell structure. This paper investigates two important factors, surface cleaning and intragrain defects elimination for the seed layer, which can greatly influence the epitaxial grown solar cell performance. Shorter incubation and crystallisation time is observed using a simplified RCA cleaning than the other two wet chemical cleaning methods, indicating a cleaner seed layer surface is achieved. Cross sectional transmission microscope images confirm a crystallographic transferal of information from the simplified RCA cleaned seed layer into the epi-layer. RTA for the SPC seed layer can effectively eliminate the intragrain defects in the seed layer and improve structural quality of both of the seed layer and the epi-layer. Consequently, epitaxial grown poly-Si solar cell on the RTA treated seed layer shows better solar cell efficiency, V{sub oc} and J{sub sc} than the one on the seed layer without RTA treatment.

Mathematical modelling of the viable epidermis: impact of cell shape and vertical arrangement

KAUST Repository

Wittum, Rebecca; Naegel, Arne; Heisig, Michael; Wittum, Gabriel

2017-01-01

In-silico methods are valuable tools for understanding the barrier function of the skin. The key benefit is that mathematical modelling allows the interplay between cell shape and function to be elucidated. This study focuses on the viable (living

Repair of Avascular Meniscus Tears with Electrospun Collagen Scaffolds Seeded with Human Cells.

Science.gov (United States)

Baek, Jihye; Sovani, Sujata; Glembotski, Nicholas E; Du, Jiang; Jin, Sungho; Grogan, Shawn P; D'Lima, Darryl D

2016-03-01

The self-healing capacity of an injured meniscus is limited to the vascularized regions and is especially challenging in the inner avascular regions. As such, we investigated the use of human meniscus cell-seeded electrospun (ES) collagen type I scaffolds to produce meniscal tissue and explored whether these cell-seeded scaffolds can be implanted to repair defects created in meniscal avascular tissue explants. Human meniscal cells (derived from vascular and avascular meniscal tissue) were seeded on ES scaffolds and cultured. Constructs were evaluated for cell viability, gene expression, and mechanical properties. To determine potential for repair of meniscal defects, human meniscus avascular cells were seeded and cultured on aligned ES collagen scaffolds for 4 weeks before implantation. Surgical defects resembling "longitudinal tears" were created in the avascular zone of bovine meniscus and implanted with cell-seeded collagen scaffolds and cultured for 3 weeks. Tissue regeneration and integration were evaluated by histology, immunohistochemistry, mechanical testing, and magentic resonance imaging. Ex vivo implantation with cell-seeded collagen scaffolds resulted in neotissue that was significantly better integrated with the native tissue than acellular collagen scaffolds or untreated defects. Human meniscal cell-seeded ES collagen scaffolds may therefore be useful in facilitating meniscal repair of avascular meniscus tears.

A combination of direct viable count and fluorescence in situ hybridization for specific enumeration of viable Lactobacillus delbrueckii subsp.bulgaricus and Streptococcus thermophilus.

Science.gov (United States)

García-Hernández, J; Moreno, Y; Amorocho, C M; Hernández, M

2012-03-01

We have developed a direct viable count (DVC)-FISH procedure for quickly and easily discriminating between viable and nonviable cells of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains, the traditional yogurt bacteria. direct viable count method has been modified and adapted for Lact. delbrueckii subsp. bulgaricus and Strep. thermophilus analysis by testing different times of incubation and concentrations of DNA-gyrase inhibitors. DVC procedure has been combined with fluorescent in situ hybridization (FISH) for the specific detection of viable cells of both bacteria with specific rRNA oligonucleotide probes (DVC-FISH). Of the four antibiotics tested (novobiocin, nalidixic acid, pipemidic acid and ciprofloxacin), novobiocin was the most effective for DVC method and the optimum incubation time was 7 h for both bacteria. The number of viable cells was obtained by the enumeration of specific hybridized cells that were elongated at least twice their original length for Lactobacillus and twice their original size for Streptococcus. This technique was successfully applied to detect viable cells in inoculated faeces. Results showed that this DVC-FISH procedure is a quick and culture-independent useful method to specifically detect viable Lact. delbrueckii subsp. bulgaricus and Strep. thermophilus in different samples, being applied for the first time to lactic acid bacteria. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

Drug Plant Seed Viability Preservation by Cryoconservation

Directory of Open Access Journals (Sweden)

Alexandra Sh. Dodonova

2013-01-01

Full Text Available The article considers the possibility of cryopreservation of seeds of several species of drug plants in Central Kazakhstan – Tanacetum ulutavicum, Niedzwedzkia semiretschenskia, Rhaponticum carthamoides. To increase the amount of viable seeds after liquid nitrogen freezing, we used different defrosting temperatures, deposited seeds with different moisture contents and used different containers for cryopreservation. Recommendations, concerning conditions of cryopreservation of seeds of these drug plant species were developed, basing on the obtained results.

High speed measurement of corn seed viability using hyperspectral imaging

Science.gov (United States)

Ambrose, Ashabahebwa; Kandpal, Lalit Mohan; Kim, Moon S.; Lee, Wang-Hee; Cho, Byoung-Kwan

2016-03-01

Corn is one of the most cultivated crops all over world as food for humans as well as animals. Optimized agronomic practices and improved technological interventions during planting, harvesting and post-harvest handling are critical to improving the quantity and quality of corn production. Seed germination and vigor are the primary determinants of high yield notwithstanding any other factors that may play during the growth period. Seed viability may be lost during storage due to unfavorable conditions e.g. moisture content and temperatures, or physical damage during mechanical processing e.g. shelling, or over heating during drying. It is therefore vital for seed companies and farmers to test and ascertain seed viability to avoid losses of any kind. This study aimed at investigating the possibility of using hyperspectral imaging (HSI) technique to discriminate viable and nonviable corn seeds. A group of corn samples were heat treated by using microwave process while a group of seeds were kept as control group (untreated). The hyperspectral images of corn seeds of both groups were captured between 400 and 2500 nm wave range. Partial least squares discriminant analysis (PLS-DA) was built for the classification of aged (heat treated) and normal (untreated) corn seeds. The model showed highest classification accuracy of 97.6% (calibration) and 95.6% (prediction) in the SWIR region of the HSI. Furthermore, the PLS-DA and binary images were capable to provide the visual information of treated and untreated corn seeds. The overall results suggest that HSI technique is accurate for classification of viable and non-viable seeds with non-destructive manner.

Pumpkin seed extract: Cell growth inhibition of hyperplastic and cancer cells, independent of steroid hormone receptors.

Science.gov (United States)

Medjakovic, Svjetlana; Hobiger, Stefanie; Ardjomand-Woelkart, Karin; Bucar, Franz; Jungbauer, Alois

2016-04-01

Pumpkin seeds have been known in folk medicine as remedy for kidney, bladder and prostate disorders since centuries. Nevertheless, pumpkin research provides insufficient data to back up traditional beliefs of ethnomedical practice. The bioactivity of a hydro-ethanolic extract of pumpkin seeds from the Styrian pumpkin, Cucurbita pepo L. subsp. pepo var. styriaca, was investigated. As pumpkin seed extracts are standardized to cucurbitin, this compound was also tested. Transactivational activity was evaluated for human androgen receptor, estrogen receptor and progesterone receptor with in vitro yeast assays. Cell viability tests with prostate cancer cells, breast cancer cells, colorectal adenocarcinoma cells and a hyperplastic cell line from benign prostate hyperplasia tissue were performed. As model for non-hyperplastic cells, effects on cell viability were tested with a human dermal fibroblast cell line (HDF-5). No transactivational activity was found for human androgen receptor, estrogen receptor and progesterone receptor, for both, extract and cucurbitin. A cell growth inhibition of ~40-50% was observed for all cell lines, with the exception of HDF-5, which showed with ~20% much lower cell growth inhibition. Given the receptor status of some cell lines, a steroid-hormone receptor independent growth inhibiting effect can be assumed. The cell growth inhibition for fast growing cells together with the cell growth inhibition of prostate-, breast- and colon cancer cells corroborates the ethnomedical use of pumpkin seeds for a treatment of benign prostate hyperplasia. Moreover, due to the lack of androgenic activity, pumpkin seed applications can be regarded as safe for the prostate. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Non-Destructive Quality Evaluation of Pepper (Capsicum annuum L. Seeds Using LED-Induced Hyperspectral Reflectance Imaging

Directory of Open Access Journals (Sweden)

Changyeun Mo

2014-04-01

Full Text Available In this study, we developed a viability evaluation method for pepper (Capsicum annuum L. seeds based on hyperspectral reflectance imaging. The reflectance spectra of pepper seeds in the 400–700 nm range are collected from hyperspectral reflectance images obtained using blue, green, and red LED illumination. A partial least squares–discriminant analysis (PLS-DA model is developed to classify viable and non-viable seeds. Four spectral ranges generated with four types of LEDs (blue, green, red, and RGB, which were pretreated using various methods, are investigated to develop the classification models. The optimal PLS-DA model based on the standard normal variate for RGB LED illumination (400–700 nm yields discrimination accuracies of 96.7% and 99.4% for viable seeds and nonviable seeds, respectively. The use of images based on the PLS-DA model with the first-order derivative of a 31.5-nm gap for red LED illumination (600–700 nm yields 100% discrimination accuracy for both viable and nonviable seeds. The results indicate that a hyperspectral imaging technique based on LED light can be potentially applied to high-quality pepper seed sorting.

Low molecular weight procyanidins from grape seeds enhance the impact of 5-Fluorouracil chemotherapy on Caco-2 human colon cancer cells.

Directory of Open Access Journals (Sweden)

Ker Y Cheah

Full Text Available OBJECTIVE: Grape seed procyanidins (PC are flavan-3-ol oligomers and polymers known for their biological activity in the gut. Grape seed extract (GSE have been reported to reduce intestinal injury in a rat model of mucositis. We sought to investigate effects of purified PC fractions differing in mean degree of polymerization (mDP combined with 5-Fluorouracil (5-FU chemotherapy on the viability of colon cancer cells (Caco-2. DESIGN: SixPC fractions (F1-F6 were isolated from Cabernet Sauvignon seeds at two ripeness stages: pre-veraison unripe (immature and ripe (mature, utilizing step gradient, low-pressure chromatography on a Sephadex LH-20 resin. Fractions were tested on Caco-2 cells, alone and in combination with 5-FU. Eluted fractions were characterized by phloroglucinolysis and gel permeation chromatography. Cell viability was determined by the 3-(4,5-Dimethylthiazol-2yl-2,5-diphenyl-tetrazolium bromide (MTT assay. RESULTS: All isolated fractions significantly reduced Caco-2 cell viability compared to the control (P<0.05, but F2 and F3 (mDP 2-6 were the most active fractions (immature F2 = 32% mDP 2.4, F3 = 35% mDP 5.8 and mature F2 = 13% mDP 3.6 and F3 = 17% mDP 5.9; percentage of viable cells remaining on Caco-2 cells. When combined with 5-FU, immature fractions F1-F3 enhanced the cell toxicity effects of 5-FU by 27-73% (P<0.05. Mature seed PC fractions (F1-F4 significantly enhanced the toxicity of 5-FU by 60-83% against Caco-2 cells (P<0.05. Moreover, some fractions alone were more potent at decreasing viability in Caco-2 cells (P<0.05; immature fractions = 65-68% and mature fractions = 83-87% compared to 5-FU alone (37%. CONCLUSIONS: PCs of mDP 2-6 (immature F1-F3 and mature F1 and F4not only enhanced the impact of 5-FU in killing Caco-2 cells, but also surpassed standard 5-FU chemotherapy as an anti-cancer agent.The bioactivity of PC is therefore attributed primarily to lower molecular weight PCs.

Viability of Macrotyloma axillare cv. Guatá seeds under chemical scarification

Directory of Open Access Journals (Sweden)

Suleize Rocha Terra

2012-12-01

Full Text Available The use of the associacion of grasses and legumes results in a good forage yield as well as a good animal performance due to maintenance of adequate levels of crude protein (CP in the animal diet, both the direct effect of the ingestion of legumes, or even through indirect effect of the introduction of nitrogen by legumes in the pasture, thereby increasing the production of forage mass. However, the seeds of legumines have the so-called hardness seeds or dormant according to their coats impermeable to water. The hardness of the seed prevents the germination of the same since these seeds have a hard shell impervious to water and gas exchange, not allowing the imbibition of water by the seed, or embryo oxygenation. This dormancy may be broken by various known methods such as scarification, promoting cracks in the seed coat allowing the entry of water therein. In the case of Macrotyloma axillare exposure of seeds to scarification method has proven to be an efficient way to break dormancy and promoting seed germination. The aim of the present work was to evaluate the viability of seeds after suffering dormancy by immersion seeds in sulfuric acid. The experiment to break dormancy Macrotyloma axillares cv. Guatá was conducted in the laboratory of Plant Physiology at the Intituto de Zootecnia in Nova Odessa, São Paulo State, using the seeds that were stored in a cold room with temperatures between 10-12 ºC, relative humidity (RH 30-33% and no light. The treatments studied were: control, chemical scarification with sulfuric acid for 5, 10, 15, 20, 30 and 40 minutes. We used a completely randomized design with two replications, being 100 seeds each repetition. After treatment the seeds were under to tetrazolium test resulting in viable seeds and seed viability. So the result of the tetrazolium test was obtained by average percentage of viable seeds, found in the repeats tested, respecting the maximum tolerances set out in Brazilian Rules for Seed

Contribution to the improvement of seed extraction in larch

Energy Technology Data Exchange (ETDEWEB)

Philippe, G. [Cemagref - Forest nurseries and Plant genetics division, Nogent-sur-Vernisson (France)

1995-12-31

Industrial techniques in a seed plant resulted in the extraction of a small percentage of seed potential (18% in hybrid larch and 40% in European larch). It was determined that the seeds remaining within the cones were as viable as the seeds extracted. Thus, the quantity of marketable seeds was directly proportional to the quantity of seeds extracted and an important gain in seed yield could be achieved by improving extraction techniques. Among the five techniques tested in a second experiment, the two cone grinding treatments, done in a hammer mill, were characterized by the best extraction rate (up to 80% of seed potential) but they resulted in a decrease in seed viability. However, cone grinding applied to large volume of cones produced highly viable seeds. The difference was attributed to the number of cones treated. With regard to other techniques, cone drying followed by cone threshing was more efficient when it was applied to cones previously cut in two or let outside for a couple of months. All these techniques gave better results when cones were collected in spring rather than in autumn whereas cone grinding efficiency did not depend on cone collection date. 8 refs, 1 fig, 2 tabs

Contribution to the improvement of seed extraction in larch

Energy Technology Data Exchange (ETDEWEB)

Philippe, G [Cemagref - Forest nurseries and Plant genetics division, Nogent-sur-Vernisson (France)

1996-12-31

Industrial techniques in a seed plant resulted in the extraction of a small percentage of seed potential (18% in hybrid larch and 40% in European larch). It was determined that the seeds remaining within the cones were as viable as the seeds extracted. Thus, the quantity of marketable seeds was directly proportional to the quantity of seeds extracted and an important gain in seed yield could be achieved by improving extraction techniques. Among the five techniques tested in a second experiment, the two cone grinding treatments, done in a hammer mill, were characterized by the best extraction rate (up to 80% of seed potential) but they resulted in a decrease in seed viability. However, cone grinding applied to large volume of cones produced highly viable seeds. The difference was attributed to the number of cones treated. With regard to other techniques, cone drying followed by cone threshing was more efficient when it was applied to cones previously cut in two or let outside for a couple of months. All these techniques gave better results when cones were collected in spring rather than in autumn whereas cone grinding efficiency did not depend on cone collection date. 8 refs, 1 fig, 2 tabs

Cell Seeding Densities in Autologous Chondrocyte Implantation Techniques for Cartilage Repair.

Science.gov (United States)

Foldager, Casper Bindzus; Gomoll, Andreas H; Lind, Martin; Spector, Myron

2012-04-01

Cartilage repair techniques have been among the most intensively investigated treatments in orthopedics for the past decade, and several different treatment modalities are currently available. Despite the extensive research effort within this field, the generation of hyaline cartilage remains a considerable challenge. There are many parameters attendant to each of the cartilage repair techniques that can affect the amount and types of reparative tissue generated in the cartilage defect, and some of the most fundamental of these parameters have yet to be fully investigated. For procedures in which in vitro-cultured autologous chondrocytes are implanted under a periosteal or synthetic membrane cover, or seeded onto a porous membrane or scaffold, little is known about how the number of cells affects the clinical outcome. Few published clinical studies address the cell seeding density that was employed. The principal objective of this review is to provide an overview of the cell seeding densities used in cell-based treatments currently available in the clinic for cartilage repair. Select preclinical studies that have informed the use of specific cell seeding densities in the clinic are also discussed.

Tolerance of Ruppia sinensis Seeds to Desiccation, Low Temperature, and High Salinity With Special Reference to Long-Term Seed Storage

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Ruiting Gu

2018-03-01

Full Text Available Seeds are important materials for the restoration of globally-threatened marine angiosperm (seagrass populations. In this study, we investigated the differences between different Ruppia sinensis seed types and developed two feasible long-term R. sinensis seed storage methods. The ability of R. sinensis seeds to tolerate the short-term desiccation and extreme cold had been investigated. The tolerance of R. sinensis seeds to long-term exposure of high salinity, cold temperature, and desiccation had been considered as potential methods for long-term seed storage. Also, three morphological and nine physiological indices were measured and compared between two types of seeds: Shape L and Shape S. We found that: (1 wet storage at a salinity of 30–40 psu and 0°C were the optimal long-term storage conditions, and the proportion of viable seeds reached over 90% after a storage period of 11 months since the seeds were collected from the reproductive shoots; (2 dry condition was not the optimal choice for long-term storage of R. sinensis seeds; however, storing seeds in a dry condition at 5°C and 33 ± 10% relative humidity for 9 months had a relatively high percentage (74.44 ± 2.22% of viable seeds, consequently desiccation exposure could also be an acceptable seed storage method; (3 R. sinensis seeds would lose vigor in the interaction of extreme cold (-27°C and desiccation; (4 there were significant differences in seed weight, seed curvature, and endocarp thickness between the two types of seeds. These findings provided fundamental physiological information for R. sinensis seeds and supported the long-term storage of its seeds. Our results may also serve as useful reference for seed storage of other threatened seagrass species and facilitate their ex situ conservation and habitat restoration.

Tolerance of Ruppia sinensis Seeds to Desiccation, Low Temperature, and High Salinity With Special Reference to Long-Term Seed Storage.

Science.gov (United States)

Gu, Ruiting; Zhou, Yi; Song, Xiaoyue; Xu, Shaochun; Zhang, Xiaomei; Lin, Haiying; Xu, Shuai; Yue, Shidong; Zhu, Shuyu

2018-01-01

Seeds are important materials for the restoration of globally-threatened marine angiosperm (seagrass) populations. In this study, we investigated the differences between different Ruppia sinensis seed types and developed two feasible long-term R. sinensis seed storage methods. The ability of R. sinensis seeds to tolerate the short-term desiccation and extreme cold had been investigated. The tolerance of R. sinensis seeds to long-term exposure of high salinity, cold temperature, and desiccation had been considered as potential methods for long-term seed storage. Also, three morphological and nine physiological indices were measured and compared between two types of seeds: Shape L and Shape S. We found that: (1) wet storage at a salinity of 30-40 psu and 0°C were the optimal long-term storage conditions, and the proportion of viable seeds reached over 90% after a storage period of 11 months since the seeds were collected from the reproductive shoots; (2) dry condition was not the optimal choice for long-term storage of R. sinensis seeds; however, storing seeds in a dry condition at 5°C and 33 ± 10% relative humidity for 9 months had a relatively high percentage (74.44 ± 2.22%) of viable seeds, consequently desiccation exposure could also be an acceptable seed storage method; (3) R. sinensis seeds would lose vigor in the interaction of extreme cold (-27°C) and desiccation; (4) there were significant differences in seed weight, seed curvature, and endocarp thickness between the two types of seeds. These findings provided fundamental physiological information for R. sinensis seeds and supported the long-term storage of its seeds. Our results may also serve as useful reference for seed storage of other threatened seagrass species and facilitate their ex situ conservation and habitat restoration.

Programmed cell death during development of cowpea (Vigna unguiculata (L.) Walp.) seed coat.

Science.gov (United States)

Lima, Nathália Bastos; Trindade, Fernanda Gomes; da Cunha, Maura; Oliveira, Antônia Elenir Amâncio; Topping, Jennifer; Lindsey, Keith; Fernandes, Kátia Valevski Sales

2015-04-01

The seed coat develops primarily from maternal tissues and comprises multiple cell layers at maturity, providing a metabolically dynamic interface between the developing embryo and the environment during embryogenesis, dormancy and germination of seeds. Seed coat development involves dramatic cellular changes, and the aim of this research was to investigate the role of programmed cell death (PCD) events during the development of seed coats of cowpea [Vigna unguiculata (L.) Walp.]. We demonstrate that cells of the developing cowpea seed coats undergo a programme of autolytic cell death, detected as cellular morphological changes in nuclei, mitochondria, chloroplasts and vacuoles, DNA fragmentation and oligonucleosome accumulation in the cytoplasm, and loss of membrane viability. We show for the first time that classes 6 and 8 caspase-like enzymes are active during seed coat development, and that these activities may be compartmentalized by translocation between vacuoles and cytoplasm during PCD events. © 2014 John Wiley & Sons Ltd.

Mag-seeding of rat bone marrow stromal cells into porous hydroxyapatite scaffolds for bone tissue engineering.

Science.gov (United States)

Shimizu, Kazunori; Ito, Akira; Honda, Hiroyuki

2007-09-01

Bone tissue engineering has been investigated as an alternative strategy for autograft transplantation. In the process of tissue engineering, cell seeding into three-dimensional (3-D) scaffolds is the first step for constructing 3-D tissues. We have proposed a methodology of cell seeding into 3-D porous scaffolds using magnetic force and magnetite nanoparticles, which we term Mag-seeding. In this study, we applied this Mag-seeding technique to bone tissue engineering using bone marrow stromal cells (BMSCs) and 3-D hydroxyapatite (HA) scaffolds. BMSCs were magnetically labeled with our original magnetite cationic liposomes (MCLs) having a positive surface charge to improve adsorption to cell surface. Magnetically labeled BMSCs were seeded onto a scaffold, and a 1-T magnet was placed under the scaffold. By using Mag-seeding, the cells were successfully seeded into the internal space of scaffolds with a high cell density. The cell seeding efficiency into HA scaffolds by Mag-seeding was approximately threefold larger than that by static-seeding (conventional method, without a magnet). After a 14-d cultivation period using the osteogenic induction medium by Mag-seeding, the level of two representative osteogenic markers (alkaline phosphatase and osteocalcin) were significantly higher than those by static-seeding. These results indicated that Mag-seeding of BMSCs into HA scaffolds is an effective approach to bone tissue engineering.

Non-destructive technique for determining the viability of soybean (Glycine max) seeds using FT-NIR spectroscopy.

Science.gov (United States)

Kusumaningrum, Dewi; Lee, Hoonsoo; Lohumi, Santosh; Mo, Changyeun; Kim, Moon S; Cho, Byoung-Kwan

2018-03-01

The viability of seeds is important for determining their quality. A high-quality seed is one that has a high capability of germination that is necessary to ensure high productivity. Hence, developing technology for the detection of seed viability is a high priority in agriculture. Fourier transform near-infrared (FT-NIR) spectroscopy is one of the most popular devices among other vibrational spectroscopies. This study aims to use FT-NIR spectroscopy to determine the viability of soybean seeds. Viable and artificial ageing seeds as non-viable soybeans were used in this research. The FT-NIR spectra of soybean seeds were collected and analysed using a partial least-squares discriminant analysis (PLS-DA) to classify viable and non-viable soybean seeds. Moreover, the variable importance in projection (VIP) method for variable selection combined with the PLS-DA was employed. The most effective wavelengths were selected by the VIP method, which selected 146 optimal variables from the full set of 1557 variables. The results demonstrated that the FT-NIR spectral analysis with the PLS-DA method that uses all variables or the selected variables showed good performance based on the high value of prediction accuracy for soybean viability with an accuracy close to 100%. Hence, FT-NIR techniques with a chemometric analysis have the potential for rapidly measuring soybean seed viability. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

PMA-Linked Fluorescence for Rapid Detection of Viable Bacterial Endospores

Science.gov (United States)

LaDuc, Myron T.; Venkateswaran, Kasthuri; Mohapatra, Bidyut

2012-01-01

The most common approach for assessing the abundance of viable bacterial endospores is the culture-based plating method. However, culture-based approaches are heavily biased and oftentimes incompatible with upstream sample processing strategies, which make viable cells/spores uncultivable. This shortcoming highlights the need for rapid molecular diagnostic tools to assess more accurately the abundance of viable spacecraft-associated microbiota, perhaps most importantly bacterial endospores. Propidium monoazide (PMA) has received a great deal of attention due to its ability to differentiate live, viable bacterial cells from dead ones. PMA gains access to the DNA of dead cells through compromised membranes. Once inside the cell, it intercalates and eventually covalently bonds with the double-helix structures upon photoactivation with visible light. The covalently bound DNA is significantly altered, and unavailable to downstream molecular-based manipulations and analyses. Microbiological samples can be treated with appropriate concentrations of PMA and exposed to visible light prior to undergoing total genomic DNA extraction, resulting in an extract comprised solely of DNA arising from viable cells. This ability to extract DNA selectively from living cells is extremely powerful, and bears great relevance to many microbiological arenas.

Evaluating the potential for weed seed dispersal based on waterfowl consumption and seed viability.

Science.gov (United States)

Farmer, Jaime A; Webb, Elisabeth B; Pierce, Robert A; Bradley, Kevin W

2017-12-01

Migratory waterfowl have often been implicated in the movement of troublesome agronomic and wetland weed species. However, minimal research has been conducted to investigate the dispersal of agronomically important weed species by waterfowl. The two objectives for this project were to determine what weed species are being consumed by ducks and snow geese, and to determine the recovery rate and viability of 13 agronomic weed species after passage through a duck's digestive system. Seed recovered from digestive tracts of 526 ducks and geese harvested during a 2-year field study had 35 020 plants emerge. A greater variety of plant species emerged from ducks each year (47 and 31 species) compared to geese (11 and 3 species). Viable seed from 11 of 13 weed species fed to ducks in a controlled feeding study were recovered. Viability rate and gut retention times indicated potential dispersal up to 2900 km from the source depending on seed characteristics and variability in waterfowl dispersal distances. Study results confirm that waterfowl are consuming seeds from a variety of agronomically important weed species, including Palmer amaranth, which can remain viable after passage through digestive tracts and have potential to be dispersed over long distances by waterfowl. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Endothelial differentiation of human stem cells seeded onto electrospun polyhydroxybutyrate/polyhydroxybutyrate-co-hydroxyvalerate fiber mesh.

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Alessandra Zonari

Full Text Available Tissue engineering is based on the association of cultured cells with structural matrices and the incorporation of signaling molecules for inducing tissue regeneration. Despite its enormous potential, tissue engineering faces a major challenge concerning the maintenance of cell viability after the implantation of the constructs. The lack of a functional vasculature within the implant compromises the delivery of nutrients to and removal of metabolites from the cells, which can lead to implant failure. In this sense, our investigation aims to develop a new strategy for enhancing vascularization in tissue engineering constructs. This study's aim was to establish a culture of human adipose tissue-derived stem cells (hASCs to evaluate the biocompatibility of electrospun fiber mesh made of polyhydroxybutyrate (PHB and its copolymer poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHB-HV and to promote the differentiation of hASCs into the endothelial lineage. Fiber mesh was produced by blending 30% PHB with 70% PHB-HV and its physical characterization was conducted using scanning electron microscopy analysis (SEM. Using electrospinning, fiber mesh was obtained with diameters ranging 300 nm to 1.3 µm. To assess the biological performance, hASCs were extracted, cultured, characterized by flow cytometry, expanded and seeded onto electrospun PHB/PHB-HV fiber mesh. Various aspects of the cells were analyzed in vitro using SEM, MTT assay and Calcein-AM staining. The in vitro evaluation demonstrated good adhesion and a normal morphology of the hASCs. After 7, 14 and 21 days of seeding hASCs onto electrospun PHB/PHB-HV fiber mesh, the cells remained viable and proliferative. Moreover, when cultured with endothelial differentiation medium (i.e., medium containing VEGF and bFGF, the hASCs expressed endothelial markers such as VE-Cadherin and the vWF factor. Therefore, the electrospun PHB/PHB-HV fiber mesh appears to be a suitable material that can be used in

Endothelial differentiation of human stem cells seeded onto electrospun polyhydroxybutyrate/polyhydroxybutyrate-co-hydroxyvalerate fiber mesh.

Science.gov (United States)

Zonari, Alessandra; Novikoff, Silviene; Electo, Naira R P; Breyner, Natália M; Gomes, Dawidson A; Martins, Albino; Neves, Nuno M; Reis, Rui L; Goes, Alfredo M

2012-01-01

Tissue engineering is based on the association of cultured cells with structural matrices and the incorporation of signaling molecules for inducing tissue regeneration. Despite its enormous potential, tissue engineering faces a major challenge concerning the maintenance of cell viability after the implantation of the constructs. The lack of a functional vasculature within the implant compromises the delivery of nutrients to and removal of metabolites from the cells, which can lead to implant failure. In this sense, our investigation aims to develop a new strategy for enhancing vascularization in tissue engineering constructs. This study's aim was to establish a culture of human adipose tissue-derived stem cells (hASCs) to evaluate the biocompatibility of electrospun fiber mesh made of polyhydroxybutyrate (PHB) and its copolymer poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHB-HV) and to promote the differentiation of hASCs into the endothelial lineage. Fiber mesh was produced by blending 30% PHB with 70% PHB-HV and its physical characterization was conducted using scanning electron microscopy analysis (SEM). Using electrospinning, fiber mesh was obtained with diameters ranging 300 nm to 1.3 µm. To assess the biological performance, hASCs were extracted, cultured, characterized by flow cytometry, expanded and seeded onto electrospun PHB/PHB-HV fiber mesh. Various aspects of the cells were analyzed in vitro using SEM, MTT assay and Calcein-AM staining. The in vitro evaluation demonstrated good adhesion and a normal morphology of the hASCs. After 7, 14 and 21 days of seeding hASCs onto electrospun PHB/PHB-HV fiber mesh, the cells remained viable and proliferative. Moreover, when cultured with endothelial differentiation medium (i.e., medium containing VEGF and bFGF), the hASCs expressed endothelial markers such as VE-Cadherin and the vWF factor. Therefore, the electrospun PHB/PHB-HV fiber mesh appears to be a suitable material that can be used in combination with

Size, physiological quality, and green seed occurrence influenced by seeding rate in soybeans

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André Sampaio Ferreira

2017-05-01

Full Text Available The seeding rate influences the intraspecific competition, which might affect the development and quality of seeds in soybean. However, the impact of seeding rate on the physical and physiological qualities of soybean seeds needs to be better elucidated. This study aimed to evaluate the effects of soybean plant density on the seed size as well as the effects of the interaction between the plant density and seed size on the seed mass, green seed occurence, and physiological seed quality. The experiments were carried out in the growing seasons of the years 2013/14 and 2014/15 in a Latossolo Vermelho distroférrico, under a randomized complete block design, using the NK 7059 RR cultivar with six replications. Four plant densities (150, 300, 440, and 560 thousand viable seeds ha–1 were evaluated. After the classification of seeds into four sizes, using a set of sieves, a 4 ×4 factorial scheme was used for the statistical analysis of the four plant densities and four seed sizes. The seed samples were evaluated for the seed mass, green seed percentage, germination, and vigor. Under thermal and water stress during seed development, an increase in the seeding rate led to a reduction in the green seed occurrence and an increase in the seed size and mass. However, in the absence of thermal and water stress, the seed size and mass were not altered by the seeding rate and, there was no occurrence of green seeds.

Development of a cell-seeded modified small intestinal submucosa for urethroplasty

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Long Zhang

2016-03-01

Conclusions: A modified 3D porous SIS scaffold seeded with UC and treated with PAA produces better urethroplasty results than cell-seeded untreated SIS scaffolds, or unseeded PAA treated SIS scaffolds.

Cytotoxic activity of kenaf (Hibiscus cannabinus L.) seed extract and oil against human cancer cell lines

Science.gov (United States)

Wong, Yu Hua; Tan, Wai Yan; Tan, Chin Ping; Long, Kamariah; Nyam, Kar Lin

2014-01-01

Objective To examine the cytotoxic properties of both the kenaf (Hibiscus cannabinus L.) seed extract and kenaf seed oil on human cervical cancer, human breast cancer, human colon cancer and human lung cancer cell lines. Methods The in vitro cytotoxic activity of the kenaf (Hibiscus cannabinus L.) seed extract and kenaf seed oil on human cancer cell lines was evaluated by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and sulforhodamine B assays. Cell morphological changes were observed by using an inverted light microscope. Results The kenaf seed extract (KSE) exhibited a lower IC50 than kenaf seed oil (KSO) in all of the cancer cell lines. Morphological alterations in the cell lines after KSE and KSO treatment were observed. KSE and KSO possessed effective cytotoxic activities against all the cell lines been selected. Conclusions KSE and KSO could be potential sources of natural anti-cancer agents. Further investigations on using kenaf seeds for anti-proliferative properties are warranted. PMID:25183141

Cranberry and Grape Seed Extracts Inhibit the Proliferative Phenotype of Oral Squamous Cell Carcinomas

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Kourt Chatelain

2011-01-01

Full Text Available Proanthocyanidins, compounds highly concentrated in dietary fruits, such as cranberries and grapes, demonstrate significant cancer prevention potential against many types of cancer. The objective of this study was to evaluate cranberry and grape seed extracts to quantitate and compare their anti-proliferative effects on the most common type of oral cancer, oral squamous cell carcinoma. Using two well-characterized oral squamous cell carcinoma cell lines, CAL27 and SCC25, assays were performed to evaluate the effects of cranberry and grape seed extract on phenotypic behaviors of these oral cancers. The proliferation of both oral cancer cell lines was significantly inhibited by the administration of cranberry and grape seed extracts, in a dose-dependent manner. In addition, key regulators of apoptosis, caspase-2 and caspase-8, were concomitantly up-regulated by these treatments. However, cranberry and grape seed extracts elicited differential effects on cell adhesion, cell morphology, and cell cycle regulatory pathways. This study represents one of the first comparative investigations of cranberry and grape seed extracts and their anti-proliferative effects on oral cancers. Previous findings using purified proanthocyanidin from grape seed extract demonstrated more prominent growth inhibition, as well as apoptosis-inducing, properties on CAL27 cells. These observations provide evidence that cranberry and grape seed extracts not only inhibit oral cancer proliferation but also that the mechanism of this inhibition may function by triggering key apoptotic regulators in these cell lines. This information will be of benefit to researchers interested in elucidating which dietary components are central to mechanisms involved in the mediation of oral carcinogenesis and progression.

STORAGE OF Handroanthus umbellatus SEEDS

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Cibele Chalita Martins

2014-09-01

Full Text Available http://dx.doi.org/10.5902/1980509815725Seed storage under controlled environmental conditions represents one of the most important lines of research to be applied on short-lived forest species as Handroanthus. The present research aimed to identify the most suitable seed storage conditions and longevity behavior of Handroanthus umbellatus seeds subject to the following storage treatments: packaging permeable paper bags under a no-controlled laboratory temperature and humidity (control and multiwall semipermeable bag at temperatures of -18 ºC, 1 ºC and 25 ºC. Seeds were dried to 6.3% of water content. Stored seeds were evaluated every three months until 24 months for water content, germination percentage and vigor utilizing first counting test. Seeds of T. umbellata are orthodox, with low longevity under natural conditions, once they remain viable for less than 5 months. The best conditions of seed preservation of these species were obtained by storage at -18° C in multiwall bags. Under these conditions physiological seed quality remains unchanged for a 24-month period.

In vitro model for human endothelial cell seeding of a small diameter vascular graft

International Nuclear Information System (INIS)

Kent, K.C.; Oshima, A.; Ikemoto, T.; Whittemore, A.D.

1988-01-01

A precise system was devised to measure the kinetics of attachment of human venous endothelium to a variety of materials and substrates. Cells were labelled in a postconfluent state with tritiated thymidine, harvested, and a cell suspension seeded into a 4 mm PTFE graft. After a 90 minute incubation period, one half of the graft segment was sacrificed and the remaining portion placed in a perfusion system (225 cc/min) for 1 hour. Graft segments, effluents, and seeding suspension were assayed in a beta scintillation counter. The percentage of cells that attached pre- and postperfusion were determined, as well as the retrieval of tritium from the system. Initially, 71% of seeded cells attached to grafts coated with fibronectin, with significantly less (60%) remaining attached after perfusion. Only 10% of cells initially attached to uncoated grafts, with 4% retained postperfusion. Retrieval of tritium averaged 102 +/- 10% for all experiments. This system determines both pre- and postperfusion attachment of human endothelial cells to vascular grafts following manipulation of numerous variables, including graft material, substrate, incubation time, and seeding density. An optimal seeding protocol for human trials can thus be determined

Plated copper front side metallization on printed seed-layers for silicon solar cells

OpenAIRE

Kraft, Achim

2015-01-01

A novel copper front side metallization architecture for silicon solar cells based on a fine printed silver seed-layer, plated with nickel, copper and silver, is investigated. The work focuses on the printing of fine seed-layers with low silver consumption, the corrosion of the printed seed-layers by the interaction with electrolyte solutions and the encapsulation material on module level and on the long term stability of the cells due to copper migration. The investigation of the correlation...

Mathematical modelling of the viable epidermis: impact of cell shape and vertical arrangement

KAUST Repository

Wittum, Rebecca

2017-12-07

In-silico methods are valuable tools for understanding the barrier function of the skin. The key benefit is that mathematical modelling allows the interplay between cell shape and function to be elucidated. This study focuses on the viable (living) epidermis. For this region, previous works suggested a diffusion model and an approximation of the cells by hexagonal prisms. The work at hand extends this in three ways. First, the extracellular space is treated with full spatial resolution. This induces a decrease of permeability by about 10%. Second, cells of tetrakaidecahedral shape are considered, in addition to the original hexagonal prisms. For both cell types, the resulting membrane permeabilities are compared. Third, for the first time, the influence of cell stacking in the vertical direction is considered. This is particularly important for the stratum granulosum, where tight junctions are present.

The healing of bony defects by cell-free collagen-based scaffolds compared to stem cell-seeded tissue engineered constructs.

LENUS (Irish Health Repository)

Lyons, Frank G

2010-12-01

One of the key challenges in tissue engineering is to understand the host response to scaffolds and engineered constructs. We present a study in which two collagen-based scaffolds developed for bone repair: a collagen-glycosaminoglycan (CG) and biomimetic collagen-calcium phosphate (CCP) scaffold, are evaluated in rat cranial defects, both cell-free and when cultured with MSCs prior to implantation. The results demonstrate that both cell-free scaffolds showed excellent healing relative to the empty defect controls and somewhat surprisingly, to the tissue engineered (MSC-seeded) constructs. Immunological analysis of the healing response showed higher M1 macrophage activity in the cell-seeded scaffolds. However, when the M2 macrophage response was analysed, both groups (MSC-seeded and non-seeded scaffolds) showed significant activity of these cells which are associated with an immunomodulatory and tissue remodelling response. Interestingly, the location of this response was confined to the construct periphery, where a capsule had formed, in the MSC-seeded groups as opposed to areas of new bone formation in the non-seeded groups. This suggests that matrix deposited by MSCs during in vitro culture may adversely affect healing by acting as a barrier to macrophage-led remodelling when implanted in vivo. This study thus improves our understanding of host response in bone tissue engineering.

Cytotoxic Effects of Alcoholic Extract of Dorema Glabrum Seed on Cancerous Cells Viability

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Maryam Bannazadeh Amirkhiz

2013-08-01

Full Text Available Purpose: In the present study cytotoxic effects of the alcoholic extract of Dorema Glabrum seed on viability of WEHI-164 cells, mouse Fibrosarcoma cell line and L929 normal cells were compared with the cytotoxic effects of Taxol (anticancer and apoptosis inducer drug. Methods: To find out the plant extract cytotoxic effects, MTT test and DNA fragmentation assay, the biochemical hallmark of apoptosis were performed on cultured and treated cells. Results: According to the findings the alcoholic extract of Dorema Glabrum seed can alter cells morphology and because of chromatin condensation and other changes they shrink and take a spherical shape, and lose their attachment too. So the plant extract inhibits cell growth albeit in a time and dose dependent manner and results in degradation of chromosomal DNA. Conclusion: Our data well established the anti-proliferative effect of methanolic extract of Dorema Glabrum seed and clearly showed that the plant extract can induce apoptosis and not necrosis in vitro, but the mechanism of its activities remained unknown. These results demonstrated that Dorema Glabrum seed might be a novel and attractive therapeutic candidate for tumor treatment in clinical practices.

Black seed oil ameliorates allergic airway inflammation by inhibiting T-cell proliferation in rats.

Science.gov (United States)

Shahzad, Muhammad; Yang, Xudong; Raza Asim, M B; Sun, Qingzhu; Han, Yan; Zhang, Fujun; Cao, Yongxiao; Lu, Shemin

2009-02-01

The black seeds, from the Ranunculaceae family, have been traditionally used by various cultures as a natural remedy for several ailments. In this study, we examined the effect of black seed oil as an immunomodulator in a rat model of allergic airway inflammation. Rats sensitized to ovalbumin and challenged intranasally with ovalbumin to induce an allergic inflammatory response were compared to ovalbumin-sensitized, intranasally ovalbumin-exposed rats pretreated with intraperitoneally administered black seed oil and to control rats. The levels of IgE, IgG1 and ova-specific T-cell proliferation in spleen were measured by ELISA. The pro-inflammatory cytokine IL-4, IL-5, IL-6 and TGF-beta1 mRNA expression levels were measured by reverse transcription polymerase chain reaction. The intraperitoneal administration of black seed oil inhibited the Th2 type immune response in rats by preventing inflammatory cell infiltration and pathological lesions in the lungs. It significantly decreased the nitric oxide production in BALF, total serum IgE, IgG1 and OVA-specific IgG1 along with IL-4, IL-5, IL-6 and TGF-beta1 mRNA expression. Black seed oil treatment resulted in decreased T-cell response evident by lesser delayed type hypersensitivity and lower T-cell proliferation in spleen. In conclusion, black seed oil exhibited a significant reduction in all the markers of allergic inflammation mainly by inhibiting the delayed type hypersensitivity and T-cell proliferation. The data suggests that inhibition of T-cell response may be responsible for immunomodulatory effect of black seed oil in the rat model of allergic airway inflammation.

9 CFR 113.27 - Detection of extraneous viable bacteria and fungi in live vaccines.

Science.gov (United States)

2010-01-01

... bacteria and fungi in live vaccines. 113.27 Section 113.27 Animals and Animal Products ANIMAL AND PLANT... bacteria and fungi in live vaccines. Unless otherwise specified by the Administrator or elsewhere exempted... Seed Bacteria shall be tested for extraneous viable bacteria and fungi as prescribed in this section. A...

Characterization of a multi-seeded (msd) mutant of sorghum that displays significant enhancement in seed number

Science.gov (United States)

Sorghum (Sorghum bicolor L. Moench) cultivars and germplasm display branched inflorescence or panicle, characterized by spikelets composed of a single sessile, fertile floret that develop into viable seed and one or two adjacent sterile pedicellate florets (Monoseeded [MSD] trait). Based on total nu...

Grazing of particle-associated bacteria-an elimination of the non-viable fraction.

Science.gov (United States)

Gonsalves, Maria-Judith; Fernandes, Sheryl Oliveira; Priya, Madasamy Lakshmi; LokaBharathi, Ponnapakkam Adikesavan

Quantification of bacteria being grazed by microzooplankton is gaining importance since they serve as energy subsidies for higher trophic levels which consequently influence fish production. Hence, grazing pressure on viable and non-viable fraction of free and particle-associated bacteria in a tropical estuary controlled mainly by protist grazers was estimated using the seawater dilution technique. In vitro incubations over a period of 42h showed that at the end of 24h, growth coefficient (k) of particle-associated bacteria was 9 times higher at 0.546 than that of free forms. Further, 'k' value of viable cells on particles was double that of free forms at 0.016 and 0.007, respectively. While bacteria associated with particles were grazed (coefficient of removal (g)=0.564), the free forms were relatively less grazed indicating that particle-associated bacteria were exposed to grazers in these waters. Among the viable and non-viable forms, 'g' of non-viable fraction (particle-associated bacteria=0.615, Free=0.0086) was much greater than the viable fraction (particle-associated bacteria=0.056, Free=0.068). Thus, grazing on viable cells was relatively low in both the free and attached states. These observations suggest that non-viable forms of particle-associated bacteria were more prone to grazing and were weeded out leaving the viable cells to replenish the bacterial standing stock. Particle colonization could thus be a temporary refuge for the "persistent variants" where the viable fraction multiply and release their progeny. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Grazing of particle-associated bacteria-an elimination of the non-viable fraction

Directory of Open Access Journals (Sweden)

Maria-Judith Gonsalves

Full Text Available Abstract Quantification of bacteria being grazed by microzooplankton is gaining importance since they serve as energy subsidies for higher trophic levels which consequently influence fish production. Hence, grazing pressure on viable and non-viable fraction of free and particle-associated bacteria in a tropical estuary controlled mainly by protist grazers was estimated using the seawater dilution technique. In vitro incubations over a period of 42 h showed that at the end of 24 h, growth coefficient (k of particle-associated bacteria was 9 times higher at 0.546 than that of free forms. Further, ‘k’ value of viable cells on particles was double that of free forms at 0.016 and 0.007, respectively. While bacteria associated with particles were grazed (coefficient of removal (g = 0.564, the free forms were relatively less grazed indicating that particle-associated bacteria were exposed to grazers in these waters. Among the viable and non-viable forms, ‘g’ of non-viable fraction (particle-associated bacteria = 0.615, Free = 0.0086 was much greater than the viable fraction (particle-associated bacteria = 0.056, Free = 0.068. Thus, grazing on viable cells was relatively low in both the free and attached states. These observations suggest that non-viable forms of particle-associated bacteria were more prone to grazing and were weeded out leaving the viable cells to replenish the bacterial standing stock. Particle colonization could thus be a temporary refuge for the “persistent variants” where the viable fraction multiply and release their progeny.

Sensitive and Specific Biomimetic Lipid Coated Microfluidics to Isolate Viable Circulating Tumor Cells and Microemboli for Cancer Detection.

Directory of Open Access Journals (Sweden)

Jia-Yang Chen

Full Text Available Here we presented a simple and effective membrane mimetic microfluidic device with antibody conjugated supported lipid bilayer (SLB "smart coating" to capture viable circulating tumor cells (CTCs and circulating tumor microemboli (CTM directly from whole blood of all stage clinical cancer patients. The non-covalently bound SLB was able to promote dynamic clustering of lipid-tethered antibodies to CTC antigens and minimized non-specific blood cells retention through its non-fouling nature. A gentle flow further flushed away loosely-bound blood cells to achieve high purity of CTCs, and a stream of air foam injected disintegrate the SLB assemblies to release intact and viable CTCs from the chip. Human blood spiked cancer cell line test showed the ~95% overall efficiency to recover both CTCs and CTMs. Live/dead assay showed that at least 86% of recovered cells maintain viability. By using 2 mL of peripheral blood, the CTCs and CTMs counts of 63 healthy and colorectal cancer donors were positively correlated with the cancer progression. In summary, a simple and effective strategy utilizing biomimetic principle was developed to retrieve viable CTCs for enumeration, molecular analysis, as well as ex vivo culture over weeks. Due to the high sensitivity and specificity, it is the first time to show the high detection rates and quantity of CTCs in non-metastatic cancer patients. This work offers the values in both early cancer detection and prognosis of CTC and provides an accurate non-invasive strategy for routine clinical investigation on CTCs.

Synthetic virus seeds for improved vaccine safety: Genetic reconstruction of poliovirus seeds for a PER.C6 cell based inactivated poliovirus vaccine.

Science.gov (United States)

Sanders, Barbara P; Edo-Matas, Diana; Papic, Natasa; Schuitemaker, Hanneke; Custers, Jerome H H V

2015-10-13

Safety of vaccines can be compromised by contamination with adventitious agents. One potential source of adventitious agents is a vaccine seed, typically derived from historic clinical isolates with poorly defined origins. Here we generated synthetic poliovirus seeds derived from chemically synthesized DNA plasmids encoding the sequence of wild-type poliovirus strains used in marketed inactivated poliovirus vaccines. The synthetic strains were phenotypically identical to wild-type polioviruses as shown by equivalent infectious titers in culture supernatant and antigenic content, even when infection cultures are scaled up to 10-25L bioreactors. Moreover, the synthetic seeds were genetically stable upon extended passaging on the PER.C6 cell culture platform. Use of synthetic seeds produced on the serum-free PER.C6 cell platform ensures a perfectly documented seed history and maximum control over starting materials. It provides an opportunity to maximize vaccine safety which increases the prospect of a vaccine end product that is free from adventitious agents. Copyright © 2015 Elsevier Ltd. All rights reserved.

Anticancer Potential of Aqueous Ethanol Seed Extract of Ziziphus mauritiana against Cancer Cell Lines and Ehrlich Ascites Carcinoma

Directory of Open Access Journals (Sweden)

Tulika Mishra

2011-01-01

Full Text Available Ziziphus mauritiana (Lamk. is a fruit tree that has folkloric implications against many ailments and diseases. In the present study, anticancer potential of seed extract of Ziziphus mauritiana in vitro against different cell lines (HL-60, Molt-4, HeLa, and normal cell line HGF by MTT assay as well as in vivo against Ehrich ascites carcinoma bearing Swiss albino mice was investigated. The extract was found to markedly inhibit the proliferation of HL-60 cells. Annexin and PI binding of treated HL-60 cells indicated apoptosis induction by extract in a dose-dependent manner. The cell cycle analysis revealed a prominent increase in sub Go population at concentration of 20 μg/ml and above. Agarose gel electrophoresis confirmed DNA fragmentation in HL-60 cells after 3 h incubation with extract. The extract also exhibited potent anticancer potential in vivo. Treatment of Ehrlich ascites carcinoma bearing Swiss albino mice with varied doses (100–800 mg/kg b.wt. of plant extract significantly reduced tumor volume and viable tumor cell count and improved haemoglobin content, RBC count, mean survival time, tumor inhibition, and percentage life span. The enhanced antioxidant status in extract-treated animals was evident from decline in levels of lipid peroxidation and increased levels of glutathione, catalase, and superoxide dismutase.

Dye Sensitized Solar Cells for Economically Viable Photovoltaic Systems.

Science.gov (United States)

Jung, Hyun Suk; Lee, Jung-Kun

2013-05-16

TiO2 nanoparticle-based dye sensitized solar cells (DSSCs) have attracted a significant level of scientific and technological interest for their potential as economically viable photovoltaic devices. While DSSCs have multiple benefits such as material abundance, a short energy payback period, constant power output, and compatibility with flexible applications, there are still several challenges that hold back large scale commercialization. Critical factors determining the future of DSSCs involve energy conversion efficiency, long-term stability, and production cost. Continuous advancement of their long-term stability suggests that state-of-the-art DSSCs will operate for over 20 years without a significant decrease in performance. Nevertheless, key questions remain in regards to energy conversion efficiency improvements and material cost reduction. In this Perspective, the present state of the field and the ongoing efforts to address the requirements of DSSCs are summarized with views on the future of DSSCs.

Locomotion during digestion changes current estimates of seed dispersal kernels by fish

NARCIS (Netherlands)

Leeuwen, van C.H.A.; Beukeboom, R.; Nolet, B.A.; Bakker, E.S.; Pollux, B.J.A.

2016-01-01

1.Dispersal of seeds by animals is an important mechanism regulating plant diversity, range expansions and invasions. Many birds, mammals, fish and reptiles regularly ingest, transport and excrete viable seeds (known as endozoochory). 2.The effectiveness of endozoochory is modelled in dispersal

Effects of γ irradiation of hydra: elimination of interstitial cells from viable hydra

International Nuclear Information System (INIS)

Fradkin, M.; Kakis, H.; Campbell, R.D.

1978-01-01

Hydra attenuata and H. magnipapillata were γ-irradiated from a cesium source. All doses which had any observable effect (3000 rad and above) resulted in a reduction in the number of interstitial cells and of their differentiated product cells, or in the complete elimination of these cells. Interstitial cells were essentially completely eliminated within 5 days after irradiation doses above 5500 rad, and these hydra died. Irradiation doses of 4200 to 5500 rad resulted in a mixture of effects: some hydra recovered completely, some lost all interstitial cells and died, and some lost interstitial cells but could be propagated, as asexually reproducing clones, by hand feeding them. Hydra of some of these hand-fed clones entirely lacked interstitial cells and did not recover interstitial cells during subsequent culturing. Yet when these hydra were repopulated by interstitial cells from a normal hydra, they were restored to normal. Nerve cells became depleted more slowly than interstitial cells following irradiation, so animals can be obtained which possess nerve but no stem (interstitial) cells. The nerve cells and other derivatives of interstitial cells eventually disappear upon prolonged culture of the hydra. Thus γ irradiation can be used to eliminate interstitial cells from hydra, leaving viable polyps composed only of epithelial cells

Fate of Salmonella enterica and Enterohemorrhagic Escherichia coli Cells Artificially Internalized into Vegetable Seeds during Germination.

Science.gov (United States)

Liu, Da; Cui, Yue; Walcott, Ronald; Chen, Jinru

2018-01-01

Vegetable seeds contaminated with bacterial pathogens have been linked to fresh-produce-associated outbreaks of gastrointestinal infections. This study was undertaken to observe the physiological behavior of Salmonella enterica and enterohemorrhagic Escherichia coli (EHEC) cells artificially internalized into vegetable seeds during the germination process. Surface-decontaminated seeds of alfalfa, fenugreek, lettuce, and tomato were vacuum-infiltrated with four individual strains of Salmonella or EHEC. Contaminated seeds were germinated at 25°C for 9 days, and different sprout/seedling tissues were microbiologically analyzed every other day. The internalization of Salmonella and EHEC cells into vegetable seeds was confirmed by the absence of pathogens in seed-rinsing water and the presence of pathogens in seed homogenates after postinternalization seed surface decontamination. Results show that 317 (62%) and 343 (67%) of the 512 collected sprout/seedling tissue samples were positive for Salmonella and EHEC, respectively. The average Salmonella populations were significantly larger ( P seed coat tissues, followed by the root tissues, but the mean EHEC populations from all sampled tissue sections were statistically similar, except in pregerminated seeds. Three Salmonella and two EHEC strains had significantly larger cell populations on sprout/seedling tissues than other strains used in the study. Salmonella and EHEC populations from fenugreek and alfalfa tissues were significantly larger than those from tomato and lettuce tissues. The study showed the fate of internalized human pathogens on germinating vegetable seeds and sprout/seedling tissues and emphasized the importance of using pathogen-free seeds for sprout production. IMPORTANCE The internalization of microorganisms into vegetable seeds could occur naturally and represents a possible pathway of vegetable seed contamination by human pathogens. The present study investigated the ability of two important

Recovery and germination of Dichrostachys cinerea seeds fed to goats (Capra hircus)

CSIR Research Space (South Africa)

Tjelele, JT

2012-01-01

Full Text Available addressed. The objective of this study was to determine the recovery rate and germination of D. cinerea seeds that pass through the digestive tract of goats. We hypothesized that 1) D. cinerea seeds will remain intact and viable after passage through...

Multispectral fluorescence imaging technique for discrimination of cucumber (Cucumis Sativus) seed viability

Science.gov (United States)

In this study, we developed a nondestructive method for discriminating viable cucumber (Cucumis sativus) seeds based on hyperspectral fluorescence imaging. The fluorescence spectra of cucumber seeds in the 420–700 nm range were extracted from hyperspectral fluorescence images obtained using 365 nm u...

Locomotion during digestion changes current estimates of seed dispersal kernels by fish

NARCIS (Netherlands)

van Leeuwen, Casper H.A.; Beukeboom, Rosanne; Nolet, Bart A.; Bakker, Elisabeth S.; Pollux, Bart J.A.

2016-01-01

Dispersal of seeds by animals is an important mechanism regulating plant diversity, range expansions and invasions. Many birds, mammals, fish, and reptiles regularly ingest, transport and excrete viable seeds (known as endozoochory). The effectiveness of endozoochory is modelled in dispersal

Study on apoptosis of prostate cancer cell induced by 125I seed irradiation

International Nuclear Information System (INIS)

Liao Anyan; Wang Junjie; Wang Jidong; Zhuang Hongqing; Zhao Yong

2007-01-01

Objective: To explore the mechanism of apoptosis induced by 125 I seed irradiation on PC3 cells. Methods: Human prostate cancer cell line PC3 was treated by irradiation of 125 I (2.77 cGy/h) with various dose. Agarose gel electrophoresis of DNA and flows cytometry were used to detect the apoptosis of PC3 cells and indirect immunofluorescence assay was used to detect the expression of Bcl-2. The activity of Caspase-3 was measured by Caspase Colorimetric Assay Kits. Results: Apoptosis of PC3 cells could be efficiently induced by 125 I seed irradiation. The apoptotic peaks were found by flow cytometry and DNA ladder appeared on 1.8% agarose gel. The activity of Caspase-3 on PC3 cells treated by 125 I seed irradiation was not changed significantly. Bcl-2 gene expression was down-regulated with the sample concentration increased. Conclusion: 125 I irradiation can induce the apoptosis of PC3 cells and the mechanism of apoptosis is related with down regulation of Bcl-2 gene expression and is not related with Caspase-3 activity. (authors)

Bone marrow cells other than stem cells seed the bone marrow after rescue transfusion of fatally irradiated mice

International Nuclear Information System (INIS)

Cronkite, E.P.; Inoue, T.; Bullis, J.E.

1987-01-01

In a previous publication, iodinated deoxyuridine ( 125 IUdR) incorporation data were interpreted as indicating that spleen colony-forming units (CFU-S) in DNA synthesis preferentially seeded bone marrow. In the present studies, the CFU-S content of marrow from irradiated, bone-marrow transfused mice was directly determined. Pretreatment of the transfused cells with cytocidal tritiated thymidine resulted in an insignificant diminution in CFU-S content when compared with nontritiated thymidine pretreatment, implying that there is no preferential seeding. The 125 IUdR incorporation data have been reinterpreted as being a result of the proliferation of other progenitor cells present that have seeded the bone marrow

Seed germination and seedling emergence of Scotch broom (Cytisus scoparius)

Science.gov (United States)

Timothy B. Harrington

2009-01-01

Scotch broom is a large, leguminous shrub that has invaded 27 U.S. states. The species produces seeds with a hard coat that remain viable in the soil for years. Growth-chamber studies were conducted to determine effects of temperature regime and cold-stratification period on seed germination. Seedling emergence, mortality, and biomass also were studied in response to...

Development of non-destructive sorting technique for viability of watermelon seed by using hyperspectral image processing

Energy Technology Data Exchange (ETDEWEB)

Bae, Hyun Jin; Seo, Young Wook; Lohumi, Santosh; Park, Eun Soo; Cho, Byoung Kwan [Biosystems Machinery Engineering, Chungnam National University, Daejeon (Korea, Republic of); Kim, Dae Yong [Logistics institude, CJ Korea Express, Seoul (Korea, Republic of)

2016-02-15

Seed viability is one of the most important parameters that is directly related with seed germination performance and seedling emergence. In this study, a hyperspectral imaging (HSI) system having a range of 1000 –2500 nm was used to classify viable watermelon seeds from nonviable seeds. In order to obtain nonviable watermelon seeds, a total of 96 seeds were artificially aged by immersing the seeds in hot water (25°C) for 15 days. Further, hyperspectral images for 192 seeds (96 normal and 96 aged) were acquired using the developed HSI system. A germination test was performed for all the 192 seeds in order to confirm their viability. Spectral data from the hyperspectral images of the seeds were extracted by selecting pixels from the region of interest. Each seed spectrum was averaged and preprocessed to develop a classification model of partial least square discriminant analysis (PLS-DA). The developed PLS-DA model showed a classification accuracy of 94.7% for the calibration set, and 84.2% for the validation set. The results demonstrate that the proposed technique can classify viable and nonviable watermelon seeds with a reasonable accuracy, and can be further converted into an online sorting system for rapid and nondestructive classification of watermelon seeds with regard to viability.

Development of non-destructive sorting technique for viability of watermelon seed by using hyperspectral image processing

International Nuclear Information System (INIS)

Bae, Hyun Jin; Seo, Young Wook; Lohumi, Santosh; Park, Eun Soo; Cho, Byoung Kwan; Kim, Dae Yong

2016-01-01

Seed viability is one of the most important parameters that is directly related with seed germination performance and seedling emergence. In this study, a hyperspectral imaging (HSI) system having a range of 1000 –2500 nm was used to classify viable watermelon seeds from nonviable seeds. In order to obtain nonviable watermelon seeds, a total of 96 seeds were artificially aged by immersing the seeds in hot water (25°C) for 15 days. Further, hyperspectral images for 192 seeds (96 normal and 96 aged) were acquired using the developed HSI system. A germination test was performed for all the 192 seeds in order to confirm their viability. Spectral data from the hyperspectral images of the seeds were extracted by selecting pixels from the region of interest. Each seed spectrum was averaged and preprocessed to develop a classification model of partial least square discriminant analysis (PLS-DA). The developed PLS-DA model showed a classification accuracy of 94.7% for the calibration set, and 84.2% for the validation set. The results demonstrate that the proposed technique can classify viable and nonviable watermelon seeds with a reasonable accuracy, and can be further converted into an online sorting system for rapid and nondestructive classification of watermelon seeds with regard to viability

Do rice suspension-cultured cells treated with abscisic acid mimic developing seeds?

Science.gov (United States)

Matsuno, Koya; Fujimura, Tatsuhito

2015-08-01

Starch synthesis is activated in the endosperm during seed development and also in rice suspension cells cultured with abscisic acid. In the anticipation that the mechanisms of starch synthesis are similar between the endosperm and the suspension cells cultured with abscisic acid, expression of genes involved in starch synthesis was evaluated in the suspension cells after abscisic acid treatment. However, it was found that the regulatory mechanism of starch synthesis in the suspension cells cultured with abscisic acid was different from that in developing seeds. Expression analyses of genes involved in oil bodies, which accumulate in the embryo and aleurone layer, and seed storage proteins, which accumulate mainly in the endosperm, showed that the former were activated in the suspension cells cultured with abscisic acid, but the latter were not. Master regulators for embryogenesis, OsVP1 (homologue of AtABI3) and OsLFL1 (homologue of AtFUS3 or AtLFL2), were expressed in the suspension cells at levels comparable to those in the embryo. From these results, it is suggested that interactions between regulators and abscisic acid control the synthesis of phytic acid and oil bodies in the cultured cells and embryo. We suggest that the system of suspension cells cultured with abscisic acid helps to reveal the mechanisms of phytic acid and oil body synthesis in embryo.

Asymbiotic in vitro seed propagation of Dendrobium.

Science.gov (United States)

Teixeira da Silva, Jaime A; Tsavkelova, Elena A; Ng, Tzi Bun; Parthibhan, S; Dobránszki, Judit; Cardoso, Jean Carlos; Rao, M V; Zeng, Songjun

2015-10-01

The ability to germinate orchids from seeds in vitro presents a useful and viable method for the propagation of valuable germplasm, maintaining the genetic heterogeneity inherent in seeds. Given the ornamental and medicinal importance of many species within the genus Dendrobium, this review explores in vitro techniques for their asymbiotic seed germination. The influence of abiotic factors (such as temperature and light), methods of sterilization, composition of basal media, and supplementation with organic additives and plant growth regulators are discussed in context to achieve successful seed germination, protocorm formation, and further seedling growth and development. This review provides both a basis for the selection of optimal conditions, and a platform for the discovery of better ones, that would allow the development of new protocols and the exploration of new hypotheses for germination and conservation of Dendrobium seeds and seedlings.

Seed germination from lowland tapir (Tapirus terrestris) fecal samples collected during the dry season in the Northern Brazilian Amazon.

Science.gov (United States)

Barcelos, Adriana Renata; Bobrowiec, Paulo Estefano D; Sanaiotti, Tânia Margarete; Gribel, Rogério

2013-03-01

This study evaluated the potential of lowland tapirs as seed dispersers in the northern Brazilian Amazon. The study analyzed the viability of seeds after passage through the gut. Fecal samples were collected from 6 different vegetation physiognomies in Viruá National Park during the dry season. The samples were then kept in a greenhouse for 16 months to allow the seeds to germinate. The seedling species were identified and classified according to the type of fruit, plant habit, seed size and type of ingestion. Of the 111 fecal samples, 94 (84.7%) had viable seeds of 75 species. Melastomataceae was the most frequent family with viable seeds in the fecal samples (69.1% of samples, N= 18 species). The data suggest that the importance of the lowland tapirs as dispersers is not restricted to the species consumed actively by frugivory but also extends to species accidentally consumed during browsing. The occurrence of both large and small viable seeds in the fecal samples as well as a number of large drupes, which probably cannot be transported via endozoochory by any other animal species, provide evidence of the ecological importance of lowland tapirs to the dynamics of the forest-campinarana vegetation mosaic in the region. © 2012 Wiley Publishing Asia Pty Ltd, ISZS and IOZ/CAS.

[Construction of a capsular tissue-engineered ureteral stent seeded with autologous urothelial cells].

Science.gov (United States)

Tan, Haisong; Fu, Weijun; Li, Jianqiang; Wang, Zhongxin; Li, Gang; Ma, Xin; Dong, Jun; Gao, Jiangping; Wang, Xiaoxiong; Zhang, Xu

2013-01-01

To investigate the feasibility of constructing a capsular poly L-lactic acid (PLLA) ureteral stent seeded with autologous urothelial cells using tissue engineering methods. The capsular ureteral stent was constructed by subcutaneously embedding PLLA ureteral stent in the back of beagles for 3 weeks to induce the formation of connective tissue on the surfaces. After decellularization of the stent, the expanded autologous urothelial cells were seeded on the stent. The surface structure and cell adhesion of the stent were observed using HE staining, scanning electron microscope (SEM) and immunocytochemical staining. MTT assay was used to evaluate urothelial cell proliferation on the capsular PLLA ureteral stent and on circumferential small intestinal submucosa graft. HE staining and VIII factor immunohistochemistry revealed numerous capillaries in the connective tissue encapsulating the stent without obvious local inflammatory response. The results of SEM and immunocytochemical staining showed that the capsule contained rich collagenic fibers forming three-dimensional structures, and the seeded autologous urothelial cells could adhere and well aligned on the surface. MTT assay showed normal growth of the cells on the stent as compared with the cells grown on circumferential small intestinal submucosa graft. The capsular PLLA ureteral stent allows adhesion and proliferation of autologous urothelial cells and shows a potential in applications of constructing tissue-engineered ureter.

Construction of ureteral grafts by seeding urothelial cells and bone marrow mesenchymal stem cells into polycaprolactone-lecithin electrospun fibers.

Science.gov (United States)

Shen, Jie; Fu, Xiaoling; Ou, Lailiang; Zhang, Min; Guan, Yong; Wang, Kai; Che, Yongzhe; Kong, Deling; Steinhof, Gustav; Li, Wenzhong; Yu, Yaoting; Ma, Nan

2010-03-01

The aim of the present study was to investigated the construction of polycaprolactone-lecithin (PCL-L) electrospun fibers as a novel scaffold material for a tissue-engineered ureter. The effect of bone marrow mesenchymal stem cells (BM-MSCs) on the neovascularization of the scaffolds and the viability of planted urothelial cells (UCs) on PCL-L were also studied. UCs were obtained from New Zealand rabbit bladders, cultured and then seeded onto the lumen of the tubular scaffolds before being subcutaneously transplanted into the space of nude mice. The cultured UCs showed vacuolar degeneration after 7 days of transplantation and they gradually degraded thereafter. To facilitate the regeneration of the tissue-engineered ureter and the survival of UCs in the implant, MSCs were seeded into the tubular grafts by rolling up the nanofibrous membrane, followed by the seeding of UCs. This facilitated the survival of the UCs, which formed several cellular layers after 30 days. The mean microvessel density was significantly increased in tissues seeded with MSCs. Cell-tracking experiments revealed that the transplanted MSCs did not integrate directly into capillaries for angiogenesis. Our results demonstrated that the PCL-L electrospun fibrous scaffold has a high potential for a tissue-engineered ureter especially when seeded with BM-MSCs, which enhanced angiogenesis.

A reliable protocol for the isolation of viable, chondrogenically differentiated human mesenchymal stem cells from high-density pellet cultures.

Science.gov (United States)

Ullah, Mujib; Hamouda, Houda; Stich, Stefan; Sittinger, Michael; Ringe, Jochen

2012-12-01

Administration of chondrogenically differentiated mesenchymal stem cells (MSC) is discussed as a promising approach for the regenerative treatment of injured or diseased cartilage. The high-density pellet culture is the standard culture for chondrogenic differentiation, but cells in pellets secrete extracellular matrix (ECM) that they become entrapped in. Protocols for cell isolation from pellets often result in cell damage and dedifferentiation towards less differentiated MSC. Therefore, our aim was to develop a reliable protocol for the isolation of viable, chondrogenically differentiated MSC from high-density pellet cultures. Human bone marrow MSC were chondrogenically stimulated with transforming growth factor-β3, and the cartilaginous structure of the pellets was verified by alcian blue staining of cartilage proteoglycans, antibody staining of cartilage collagen type II, and quantitative real-time reverse-transcription polymerase chain reaction of the marker genes COL2A1 and SOX9. Trypsin and collagenases II and P were tested alone or in combination, and for different concentrations and times, to find a protocol for optimized pellet digestion. Whereas trypsin was not able to release viable cells, 90-min digestion with 300 U of collagenase II, 20 U of collagenase P, and 2 mM CaCl2 worked quite well and resulted in about 2.5×10(5) cells/pellet. The protocol was further optimized for the separation of released cells and ECM from each other. Cells were alcian blue and collagen type II positive and expressed COL2A1 and SOX9, verifying a chondrogenic character. However, they had different morphological shapes. The ECM was also uniformly alcian blue and collagen type II positive but showed different organizational and structural forms. To conclude, our protocol allows the reliable isolation of a defined number of viable, chondrogenically differentiated MSC from high-density pellet cultures. Such cells, as well as the ECM components, are of interest as

PMA-PhyloChip DNA Microarray to Elucidate Viable Microbial Community Structure

Science.gov (United States)

Venkateswaran, Kasthuri J.; Stam, Christina N.; Andersen, Gary L.; DeSantis, Todd

2011-01-01

Since the Viking missions in the mid-1970s, traditional culture-based methods have been used for microbial enumeration by various NASA programs. Viable microbes are of particular concern for spacecraft cleanliness, for forward contamination of extraterrestrial bodies (proliferation of microbes), and for crew health/safety (viable pathogenic microbes). However, a "true" estimation of viable microbial population and differentiation from their dead cells using the most sensitive molecular methods is a challenge, because of the stability of DNA from dead cells. The goal of this research is to evaluate a rapid and sensitive microbial detection concept that will selectively estimate viable microbes. Nucleic acid amplification approaches such as the polymerase chain reaction (PCR) have shown promise for reducing time to detection for a wide range of applications. The proposed method is based on the use of a fluorescent DNA intercalating agent, propidium monoazide (PMA), which can only penetrate the membrane of dead cells. The PMA-quenched reaction mixtures can be screened, where only the DNA from live cells will be available for subsequent PCR reaction and microarray detection, and be identified as part of the viable microbial community. An additional advantage of the proposed rapid method is that it will detect viable microbes and differentiate from dead cells in only a few hours, as opposed to less comprehensive culture-based assays, which take days to complete. This novel combination approach is called the PMA-Microarray method. DNA intercalating agents such as PMA have previously been used to selectively distinguish between viable and dead bacterial cells. Once in the cell, the dye intercalates with the DNA and, upon photolysis under visible light, produces stable DNA adducts. DNA cross-linked in this way is unavailable for PCR. Environmental samples suspected of containing a mixture of live and dead microbial cells/spores will be treated with PMA, and then incubated

Fabrication of dye-sensitized solar cell (DSSC) using annato seeds (Bixa orellana Linn)

Energy Technology Data Exchange (ETDEWEB)

Haryanto, Ditia Allindira; Landuma, Suarni; Purwanto, Agus [Department of Chemical Engineering, Sebelas Maret University, Surakarta 632112 (Indonesia)

2014-02-24

The Fabrication of dye sensitized solar cell (DSSC) using Annato seeds has been conducted in this study. Annato seeds (Bixa orellana Linn) used as a sensitizer for dye sensitized solar cell. The experimental parameter was concentration of natural dye. Annato seeds was extracted using etanol solution and the concentration was controlled by varying mass of Annato seeds. A semiconductor TiO{sub 2} was prepared by a screen printing method for coating glass use paste of TiO{sub 2}. Construction DSSC used layered systems (sandwich) consists of working electrode (TiO{sub 2} semiconductor-dye) and counter electrode (platina). Both are placed on conductive glass and electrolytes that occur electrons cycle. The characterization of thin layer of TiO{sub 2} was conducted using SEM (Scanning Electron Microscpy) analysis showed the surface morphology of TiO{sub 2} thin layer and the cross section of a thin layer of TiO{sub 2} with a thickness of 15–19 μm. Characterization of natural dye extract was determined using UV-Vis spectrometry analysis shows the wavelength range annato seeds is 328–515 nm, and the voltage (V{sub oc}) and electric current (I{sub sc}) resulted in keithley test for 30 gram, 40 gram, and 50 gram were 0,4000 V; 0,4251 V; 0,4502 V and 0,000074 A; 0,000458 A; 0,000857 A, respectively. The efficiencies of the fabricated solar cells using annato seeds as senstizer for each varying mass are 0,00799%, 0,01237%, and 0,05696%.

Characterization and Schwann Cell Seeding of up to 15.0 cm Long Spider Silk Nerve Conduits for Reconstruction of Peripheral Nerve Defects

Directory of Open Access Journals (Sweden)

Tim Kornfeld

2016-11-01

Full Text Available Nerve reconstruction of extended nerve defect injuries still remains challenging with respect to therapeutic options. The gold standard in nerve surgery is the autologous nerve graft. Due to the limitation of adequate donor nerves, surgical alternatives are needed. Nerve grafts made out of either natural or artificial materials represent this alternative. Several biomaterials are being explored and preclinical and clinical applications are ongoing. Unfortunately, nerve conduits with successful enhancement of axonal regeneration for nerve defects measuring over 4.0 cm are sparse and no conduits are available for nerve defects extending to 10.0 cm. In this study, spider silk nerve conduits seeded with Schwann cells were investigated for in vitro regeneration on defects measuring 4.0 cm, 10.0 cm and 15.0 cm in length. Schwann cells (SCs were isolated, cultured and purified. Cell purity was determined by immunofluorescence. Nerve grafts were constructed out of spider silk from Nephila edulis and decellularized ovine vessels. Finally, spider silk implants were seeded with purified Schwann cells. Cell attachment was observed within the first hour. After 7 and 21 days of culture, immunofluorescence for viability and determination of Schwann cell proliferation and migration throughout the conduits was performed. Analyses revealed that SCs maintained viable (>95% throughout the conduits independent of construct length. SC proliferation on the spider silk was determined from day 7 to day 21 with a proliferation index of 49.42% arithmetically averaged over all conduits. This indicates that spider silk nerve conduits represent a favorable environment for SC attachment, proliferation and distribution over a distance of least 15.0 cm in vitro. Thus spider silk nerve implants are a highly adequate biomaterial for nerve reconstruction.

Fertilization-independent seed development in Arabidopsis thaliana

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Chaudhury, Abdul M.; Ming, Luo; Miller, Celia; Craig, Stuart; Dennis, Elizabeth S.; Peacock, W. James

1997-01-01

We report mutants in Arabidopsis thaliana (fertilization-independent seed: fis) in which certain processes of seed development are uncoupled from the double fertilization event that occurs after pollination. These mutants were isolated as ethyl methanesulfonate-induced pseudo-revertants of the pistillata phenotype. Although the pistillata (pi) mutant has short siliques devoid of seed, the fis mutants in the pi background have long siliques containing developing seeds, even though the flowers remain free of pollen. The three fis mutations map to loci on three different chromosomes. In fis1 and fis2 seeds, the autonomous endosperm nuclei are diploid and the endosperm develops to the point of cellularization; the partially developed seeds then atrophy. In these two mutants, proembryos are formed in a low proportion of seeds and do not develop beyond the globular stage. When FIS/fis plants are pollinated by pollen from FIS/FIS plants, ≈50% of the resulting seeds contain fully developed embryos; these seeds germinate and form viable seedlings (FIS/FIS). The other 50% of seeds shrivel and do not germinate; they contain embryos arrested at the torpedo stage (FIS/fis). In normal sexual reproduction, the products of the FIS genes are likely to play important regulatory roles in the development of seed after fertilization. PMID:9108133

Fertilization-independent seed development in Arabidopsis thaliana.

Science.gov (United States)

Chaudhury, A M; Ming, L; Miller, C; Craig, S; Dennis, E S; Peacock, W J

1997-04-15

We report mutants in Arabidopsis thaliana (fertilization-independent seed:fis) in which certain processes of seed development are uncoupled from the double fertilization event that occurs after pollination. These mutants were isolated as ethyl methanesulfonate-induced pseudo-revertants of the pistillata phenotype. Although the pistillata (pi) mutant has short siliques devoid of seed, the fis mutants in the pi background have long siliques containing developing seeds, even though the flowers remain free of pollen. The three fis mutations map to loci on three different chromosomes. In fis1 and fis2 seeds, the autonomous endosperm nuclei are diploid and the endosperm develops to the point of cellularization; the partially developed seeds then atrophy. In these two mutants, proembryos are formed in a low proportion of seeds and do not develop beyond the globular stage. When FIS/fis plants are pollinated by pollen from FIS/FIS plants, approximately 50% of the resulting seeds contain fully developed embryos; these seeds germinate and form viable seedlings (FIS/FIS). The other 50% of seeds shrivel and do not germinate; they contain embryos arrested at the torpedo stage (FIS/fis). In normal sexual reproduction, the products of the FIS genes are likely to play important regulatory roles in the development of seed after fertilization.

Comparacion de la viabilidad de las semillas obtenidas por medio de las tecnicas de extraccion fisica por lavado y de germinacion Comparing the viability of seeds obtained by the physical extraction techniques VBP and VBP plus germination

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M.S. Zuluaga

2004-06-01

Full Text Available El objetivo del presente trabajo fue comparar la viabilidad obtenida por la técnica de viabilidad por presión (VPP respecto de la viabilidad obtenida por una combinación de la metodología VPP más germinación. Asimismo se trató de establecer si ambos procedimientos determinan la viabilidad de las mismas especies o si existen diferencias en las especies establecidas como viables. El número de semillas viables estimado por la técnica VPP fue de 3475 (9,85% del total del banco de semillas, en tanto el número de semillas no viables fue de 31795 (90,15% del total del banco de semillas. Luego de aplicar la técnica de germinación (5 ciclos sobre ambas categorías, en las viables se obtuvo una germinación de 3286 semillas (9,32% del total del banco de semillas, no germinando 189 semillas (0,53% del total de dicho banco. Hubo un 7.23% adicional del banco de semillas que resultó viable por germinación, cuando previamente había sido considerado no viable por la técnica VPP. Los resultados alcanzados muestran que la técnica VPP realiza una subestimación significativa del banco de semillas, en tanto la sobreestimación no alcanzó niveles de significancia. Estos resultados ponen de manifiesto que con la técnica VPP no sólo se obtiene un menor número total de semillas viables, sino que no permitiría reflejar la verdadera proporción de semillas viables y no viables del banco total. Cuando se incorpora al análisis la técnica de germinación, se encuentra que la proporción del banco no es la misma de acuerdo a la metodología empleada, pues mientras la VPP establece, dentro del banco, semillas viables y no viables, se observó que aplicando la técnica de germinación algunas semillas consideradas viables no germinaron mientras que otras consideradas como no viables sí lo hiciero.The objective of this study was to compare the viability of seeds obtained through the techniques "viability by pressure" (VBP and "viability by pressure plus

Effect of 125I seeds and 103Pd stents on proliferation of vascular smooth muscle cells

International Nuclear Information System (INIS)

Zhu Jun; Zhu Ruisen

2004-01-01

To establish the theoretical and practical base for implementing radioactive stents aft PTCA in order to prevent restenosis, in vitro observation was taken over the effects of 12 '5I-seeds and 103 Pd-implanted stents on the vascular smooth muscle cell (VSMC) proliferation. In vitro VSMC model from guinea-pig aortic arteries was established using adherent cell culture methods. The effects of 125 I-seeds and 103 Pd-implanted stents on the VSMC proliferation, with or without fetal bovine serum (FCS), were investigated through cell counting methods and 3 H-TDR implementation tests. It was shown that (1) 10% FCS significantly promoted the DNA synthesis of VSMC (P 125 I-seeds and 103 Pd-implanted stents inhibited the VSMC DNA synthesis in dose-dependent manner, regardless of 10% FCS inducement. At lower radioactive doses, neither 125 I-seeds (18.5-74 kBq) nor 103 Pd-implanted stents (1.48-2.96 MBq) exhibited distinctive effects on the VSMC DNA synthesis (P>0.05); and (3) 48 hour exposure from 125 I-seeds at 128 kBq or 10 '3Pd-implanted stents at 7.4 MBq did not result in VSMC morphological alteration, but 125 I-seeds at 370 kBq caused cells' morphological changes. Therefore both 125 I-seeds and 103 Pd-implanted stents inhibit the in vitro VSMC DNA synthesis, and the inhibition effects are significantly related to their exposure duration and doses. (authors)

Viability of small seeds found in feces of the Central American tapir on Barro Colorado Island, Panama

NARCIS (Netherlands)

Capece, P.I.; Aliaga-Rossel, E.; Jansen, P.A.

2013-01-01

Tapirs are known as effective dispersers of large-seeded tree species, but their role in dispersing small-seeded plant species has yet to be established. Tapir feces have been reported to contain large numbers of small seeds, but whether these are viable has rarely been evaluated. We determined the

Seed longevity of Eragrostis plana Nees buried in natural grassland soil

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Renato Borges de Medeiros

2014-11-01

Full Text Available The objective of this research was to evaluate the seed longevity of Eragrostis plana Nees buried at different soil depths, in a natural-grassland area in the Pampa biome (46 m altitude, 30º05´S and 51º40´W of Rio Grande do Sul State, Brazil. The experimental design was a split-plot type in complete blocks with two factors: seeds buried at five different depth levels (soil surface and 2.5, 5, 10 and 20 cm and seven exhumation dates. The blocks were allocated in natural grassland grazed by cattle, allocated in a 12-m-long transection. Fifty-four permeable nylon bags filled with 100 seeds in each division, with five vertical divisions, were buried in each row. Seven exhumation dates were used: the first on October 14, 2003 and the last on January 14, 2006. The percentage of viable seeds of E. plana, collected at seven exhumation times and set at different depths in the soil horizon, were described by simple negative exponential equations. Based on the model, the percentage of viable seeds collected at the five depths, (soil surface and 2.5, 5, 10, and 20 cm, after 2.5 years of burial, were 0.1, 0.5, 1.0, 7.4 and 22.1%, respectively. Increase in depth is directly associated with physical and physiological seed integrity of E. plana. Negative simple exponential equations can be used to predict seed longevity of E. plana buried in nylon bags. This invader species accumulates soil seed-bank of high longevity.

Real-time monitoring of non-viable airborne particles correlates with airborne colonies and represents an acceptable surrogate for daily assessment of cell-processing cleanroom performance.

Science.gov (United States)

Raval, Jay S; Koch, Eileen; Donnenberg, Albert D

2012-10-01

Airborne particulate monitoring is mandated as a component of good manufacturing practice. We present a procedure developed to monitor and interpret airborne particulates in an International Organization for Standardization (ISO) class 7 cleanroom used for the cell processing of Section 351 and Section 361 products. We collected paired viable and non-viable airborne particle data over a period of 1 year in locations chosen to provide a range of air quality. We used receiver operator characteristic (ROC) analysis to determine empirically the relationship between non-viable and viable airborne particle counts. Viable and non-viable particles were well-correlated (r(2) = 0.78), with outlier observations at the low end of the scale (non-viable particles without detectable airborne colonies). ROC analysis predicted viable counts ≥ 0.5/feet(3) (a limit set by the United States Pharmacopeia) at an action limit of ≥ 32 000 particles (≥ 0.5 µ)/feet(3), with 95.6% sensitivity and 50% specificity. This limit was exceeded 2.6 times during 18 months of retrospective daily cleanroom data (an expected false alarm rate of 1.3 times/year). After implementing this action limit, we were alerted in real time to an air-handling failure undetected by our hospital facilities management. A rational action limit for non-viable particles was determined based on the correlation with airborne colonies. Reaching or exceeding the action limit of 32 000 non-viable particles/feet(3) triggers suspension of cleanroom cell-processing activities, deep cleaning, investigation of air handling, and a deviation management process. Our full procedure for particle monitoring is available as an online supplement.

Seed germination, seedling traits, and seed bank of the tree Moringa peregrina (Moringaceae) in a hyper-arid environment.

Science.gov (United States)

Gomaa, Nasr H; Picó, F Xavier

2011-06-01

Water-limited hot environments are good examples of hyper-aridity. Trees are scarce in these environments but some manage to survive, such as the tree Moringa peregrina. Understanding how trees maintain viable populations in extremely arid environments may provide insight into the adaptive mechanisms by which trees cope with extremely arid weather conditions. This understanding is relevant to the current increasing aridity in several regions of the world. Seed germination experiments were conducted to assess variation in seed mass, seed germination, and seedling traits of Moringa peregrina plants and the correlations among these traits. A seed burial experiment was also designed to study the fate of M. peregrina seeds buried at two depths in the soil for two time periods. On average, seeds germinated in three days and seedling shoots grew 0.7 cm per day over three weeks. Larger seeds decreased germination time and increased seedling growth rates relative to smaller seeds. Seeds remained quiescent in the soil and germination was very high at both depths and burial times. The after-ripening time of Moringa peregrina seeds is short and seeds germinate quickly after imbibition. Plants of M. peregrina may increase in hyper-arid environments from seeds with larger mass, shorter germination times, and faster seedling growth rates. The results also illustrate the adjustment in allocation to seed biomass and correlations among seed and seedling traits that allows M. peregrina to be successful in coping with aridity in its environment.

Storage of sunflower seeds

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Denise de Castro Lima

Full Text Available The sunflower is among the top five crops in the world for the production of edible vegetable oil. The species displays rustic behavior, with an excellent edaphic and climatic adaptability index, being able to be cultivated throughout Brazil. Seed quality is the key to increasing production and productivity in the sunflower. The objective of this work was to monitor the viability of sunflower seeds with a view to their conservation when stored in different environments and packaging. The seeds were packed in paper bags, multilayered paper, black polyethylene and PET bottles; and stored for a period of twelve months in the following environments: dry cold room (10 ºC and 55% RH, the ambient conditions of Fortaleza, Ceará, Brazil (30-32 ºC and 75% RH, refrigerator (4 ºC and 38-43% RH and freezer (-20 ºC. Every three months, the water content of the seeds was determined and germination, accelerated ageing, speed of emergence index, and seedling dry weight were evaluated. The experimental design was completely randomized, in a scheme of split-lots, with four replications. It can be concluded that the natural environment is not suitable for the storage of sunflower seeds. Sunflower seeds remain viable for 12 months when stored in a dry cold room, refrigerator or freezer, irrespective of the type of packaging used.

Measurements and simulations of seeded electron microbunches with collective effects

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K. Hacker

2015-09-01

Full Text Available Measurements of the longitudinal phase-space distributions of electron bunches seeded with an external laser were done in order to study the impact of collective effects on seeded microbunches in free-electron lasers. When the collective effects of Coulomb forces in a drift space and coherent synchrotron radiation in a chicane are considered, velocity bunching of a seeded microbunch appears to be a viable alternative to compression with a magnetic chicane under high-gain harmonic generation seeding conditions. Measurements of these effects on seeded electron microbunches were performed with a rf deflecting structure and a dipole magnet which streak out the electron bunch for single-shot images of the longitudinal phase-space distribution. Particle tracking simulations in 3D predicted the compression dynamics of the seeded microbunches with collective effects.

Viable Cancer Cells in the Remnant Stomach are a Potential Source of Peritoneal Metastasis after Curative Distal Gastrectomy for Gastric Cancer.

Science.gov (United States)

Murata, Satoshi; Yamamoto, Hiroshi; Yamaguchi, Tsuyoshi; Kaida, Sachiko; Ishida, Mitsuaki; Kodama, Hirokazu; Takebayashi, Katsushi; Shimizu, Tomoharu; Miyake, Toru; Tani, Tohru; Kushima, Ryoji; Tani, Masaji

2016-09-01

The mechanisms underlying peritoneal metastasis (PM) after curative gastrectomy for gastric cancer (GC) are not well elucidated. This study assessed whether viable cancer cells, including cancer stemlike cells (CSCs), were present in the remnant stomach immediately before gastrointestinal (GI) tract reconstruction because these could be a source of PM after gastrectomy. Saline fluid used for remnant stomach lumen irrigation before GI reconstruction was prospectively collected from 142 consecutive patients undergoing distal gastrectomy for GC and cytologically examined. Proliferative activity (Ki67 staining) and stemness (expression of the CSC surface markers CD44s or CD44v6) were evaluated in detected cancer cells. Viable cancer cells were detected in 33 (23.2 %) of the 142 remnant stomachs. These cells formed clusters and stained positively for Ki67, indicating proliferation. Cancer cells in remnant stomachs and surface cancer cells in primary GCs from 10 (30.3 %) of these 33 cases also stained positively for CD44s or CD44v6. In a multiple logistic regression analysis, advanced cancer (odds ratio [OR], 4.65; 95 % confidence interval [CI], 1.32-16.4; P = 0.017), tumor size of 40 mm or larger (OR, 3.78; 95 % CI, 1.12-12.8; P = 0.033), and histologic differentiation (OR, 3.10; 95 % CI, 1.30-7.40; P = 0.011) were associated independently with the presence of cancer cells in the remnant stomach. Viable, proliferative, and clustered cancer cells, including CSCs, were found in remnant gastric lumens immediately before GI reconstruction, indicating a possible cellular source of PM after curative gastrectomy for GC. Dissemination of gastric contents into the peritoneal cavity should be avoided during GI reconstruction.

Bone repair by periodontal ligament stem cell-seeded nanohydroxyapatite-chitosan scaffold

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Ge S

2012-10-01

Full Text Available Shaohua Ge,1 Ning Zhao,1 Lu Wang,1 Meijiao Yu,1 Hong Liu,2 Aimei Song,1 Jing Huang,1 Guancong Wang,2 Pishan Yang11Key Laboratory of Oral Biomedicine of Shandong Province, Department of Periodontology, School of Stomatology, 2Center of Bio and Micro/Nano Functional Materials, State Key Laboratory of Crystal Materials, Shandong University, Jinan, ChinaBackground: A nanohydroxyapatite-coated chitosan scaffold has been developed in recent years, but the effect of this composite scaffold on the viability and differentiation of periodontal ligament stem cells (PDLSCs and bone repair is still unknown. This study explored the behavior of PDLSCs on a new nanohydroxyapatite-coated genipin-chitosan conjunction scaffold (HGCCS in vitro as compared with an uncoated genipin-chitosan framework, and evaluated the effect of PDLSC-seeded HGCCS on bone repair in vivo.Methods: Human PDLSCs were cultured and identified, seeded on a HGCCS and on a genipin-chitosan framework, and assessed by scanning electron microscopy, confocal laser scanning microscopy, MTT, alkaline phosphatase activity, and quantitative real-time polymerase chain reaction at different time intervals. Moreover, PDLSC-seeded scaffolds were used in a rat calvarial defect model, and new bone formation was assessed by hematoxylin and eosin staining at 12 weeks postoperatively.Results: PDLSCs were clonogenic and positive for STRO-1. They had the capacity to undergo osteogenic and adipogenic differentiation in vitro. When seeded on HGCCS, PDLSCs exhibited significantly greater viability, alkaline phosphatase activity, and upregulated the bone-related markers, bone sialoprotein, osteopontin, and osteocalcin to a greater extent compared with PDLSCs seeded on the genipin-chitosan framework. The use of PDLSC-seeded HGCCS promoted calvarial bone repair.Conclusion: This study demonstrates the potential of HGCCS combined with PDLSCs as a promising tool for bone regeneration.Keywords: periodontal ligament, stem

Tumor-selective replication herpes simplex virus-based technology significantly improves clinical detection and prognostication of viable circulating tumor cells

DEFF Research Database (Denmark)

Zhang, Wen; Bao, Li; Yang, Shaoxing

2016-01-01

Detection of circulating tumor cells remains a significant challenge due to their vast physical and biological heterogeneity. We developed a cell-surface-marker-independent technology based on telomerase-specific, replication-selective oncolytic herpes-simplex-virus-1 that targets telomerase......-reverse-transcriptase-positive cancer cells and expresses green-fluorescent-protein that identifies viable CTCs from a broad spectrum of malignancies. Our method recovered 75.5-87.2% of tumor cells spiked into healthy donor blood, as validated by different methods, including single cell sequencing. CTCs were detected in 59-100% of 326...

Seed Germination of selected Taxa from Kachchh Desert, India

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Vinay Madhukar RAOLE

2010-06-01

Full Text Available The district of Kachchh contains many culturally important plants. However, their conservation status is little known due to direct and indirect human activities. This study was undertaken with the aim of contributing to the conservation of the native species of these semi-arid regions through germination trials under laboratory conditions. Mature fruits of ten selected species were collected randomly from the known habitats to obtain viable seeds. These seeds were pre-treated with growth regulators singly or in combination after acid scarification or without scarification. Seeds were found to be dormant due to presence of thick seed coat or due to low level of endogenous hormonal level. Most of these seeds required different storage period to mature. Only seeds of Capparis cartilaginea germinated without treatment while the other species required treatments. Addition of growth regulators has enhanced seed germination in few taxa singly and in some plant cases in combination.

Clinically viable magnetic poly(lactide-co-glycolide) (PLGA) particles for MRI-based cell tracking

Science.gov (United States)

Granot, Dorit; Nkansah, Michael K.; Bennewitz, Margaret F.; Tang, Kevin S.; Markakis, Eleni A.; Shapiro, Erik M.

2013-01-01

Purpose To design, fabricate, characterize and in vivo assay clinically viable magnetic particles for MRI-based cell tracking. Methods PLGA encapsulated magnetic nano- and microparticles were fabricated. Multiple biologically relevant experiments were performed to assess cell viability, cellular performance and stem cell differentiation. In vivo MRI experiments were performed to separately test cell transplantation and cell migration paradigms, as well as in vivo biodegradation. Results Highly magnetic nano- (~100 nm) and microparticles (~1–2 μm) were fabricated. Magnetic cell labeling in culture occurred rapidly achieving 3–50 pg Fe/cell at 3 hrs for different particles types, and >100 pg Fe/cell after 10 hours, without the requirement of a transfection agent, and with no effect on cell viability. The capability of magnetically labeled mesenchymal or neural stem cells to differentiate down multiple lineages, or for magnetically labeled immune cells to release cytokines following stimulation, was uncompromised. An in vivo biodegradation study revealed that NPs degraded ~80% over the course of 12 weeks. MRI detected as few as 10 magnetically labeled cells, transplanted into the brains of rats. Also, these particles enabled the in vivo monitoring of endogenous neural progenitor cell migration in rat brains over 2 weeks. Conclusion The robust MRI properties and benign safety profile of these particles make them promising candidates for clinical translation for MRI-based cell tracking. PMID:23568825

Beneficial Effect of Jojoba Seed Extracts on Hyperglycemia-Induced Oxidative Stress in RINm5f Beta Cells.

Science.gov (United States)

Belhadj, Sahla; Hentati, Olfa; Hamdaoui, Ghaith; Fakhreddine, Khaskhoussi; Maillard, Elisa; Dal, Stéphanie; Sigrist, Séverine

2018-03-20

Hyperglycemia occurs during diabetes and insulin resistance. It causes oxidative stress by increasing reactive oxygen species (ROS) levels, leading to cellular damage. Polyphenols play a central role in defense against oxidative stress. In our study, we investigated the antioxidant properties of simmondsin, a pure molecule present in jojoba seeds, and of the aqueous extract of jojoba seeds on fructose-induced oxidative stress in RINm5f beta cells. The exposure of RINm5f beta cells to fructose triggered the loss of cell viability (-48%, p jojoba seed extract makes jojoba a powerful agent to prevent the destruction of RINm5f beta cells induced by hyperglycemia.

Alpha-synuclein cell-to-cell transfer and seeding in grafted dopaminergic neurons in vivo.

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Elodie Angot

Full Text Available Several people with Parkinson's disease have been treated with intrastriatal grafts of fetal dopaminergic neurons. Following autopsy, 10-22 years after surgery, some of the grafted neurons contained Lewy bodies similar to those observed in the host brain. Numerous studies have attempted to explain these findings in cell and animal models. In cell culture, α-synuclein has been found to transfer from one cell to another, via mechanisms that include exosomal transport and endocytosis, and in certain cases seed aggregation in the recipient cell. In animal models, transfer of α-synuclein from host brain cells to grafted neurons has been shown, but the reported frequency of the event has been relatively low and little is known about the underlying mechanisms as well as the fate of the transferred α-synuclein. We now demonstrate frequent transfer of α-synuclein from a rat brain engineered to overexpress human α-synuclein to grafted dopaminergic neurons. Further, we show that this model can be used to explore mechanisms underlying cell-to-cell transfer of α-synuclein. Thus, we present evidence both for the involvement of endocytosis in α-synuclein uptake in vivo, and for seeding of aggregation of endogenous α-synuclein in the recipient neuron by the transferred α-synuclein. Finally, we show that, at least in a subset of the studied cells, the transmitted α-synuclein is sensitive to proteinase K. Our new model system could be used to test compounds that inhibit cell-to-cell transfer of α-synuclein and therefore might retard progression of Parkinson neuropathology.

Differentially expressed genes during the imbibition of dormant and after-ripened seeds - a reverse genetics approach

NARCIS (Netherlands)

Yazdanpanah, Farzaneh; Hanson, Johannes; Hilhorst, Henk W.M.; Bentsink, Leónie

2017-01-01

Background: Seed dormancy, defined as the incapability of a viable seed to germinate under favourable conditions, is an important trait in nature and agriculture. Despite extensive research on dormancy and germination, many questions about the molecular mechanisms controlling these traits remain

Cultivation of murine bone marrow macrophages in sponges: a method that permits recovery of viable cultured cells

Energy Technology Data Exchange (ETDEWEB)

Akporiaye, E T; Stewart, S; Stewart, C C

1984-01-01

Various investigators have cultured murine bone marrow or peritoneal cells in vitro on glass or plastic surfaces with the ultimate aim of retrieving adherent macrophages for morphologic and functional evaluation. The removal of these adherent macrophages by conventional techniques has been consistently accompanied by low yield and significant cell damage. The authors report here a simple technique for culturing murine bone marrow cells in gelatin sponges (Spongostan and Gelfoam) in growth medium containing 10% fetal bovine serum and 10% L-cell conditioned medium. Viable cells were retrieved from the sponges in 10 min by digestion with collagenase. The in situ growth kinetics were similar to those found for cells cultured on plastic dishes. The recovered cells were adherent, phagocytic, positive for Fc ..gamma.. receptors, and had esterase activity. 23 references, 1 figure, 1 table.

Spillage of Viable Seeds of Oilseed Rape along Transportation Routes: Ecological Risk Assessment and Perspectives on Management Efforts

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Kathrin Pascher

2017-09-01

Full Text Available Seed spillage during handling and transportation promotes establishment and invasion of feral crops into adjacent semi-natural habitats. This is also the case for oilseed rape (OSR, Brassica napus, where seed spillage may lead to establishment of herbicide resistant OSR populations in countries without cultivation of genetically modified OSR. Using data from Austria—where cultivation and import of genetically modified OSR are banned—as a prime example, we demonstrate that ports, oil mills, switchyards, and border railway stations to countries with different electric current systems—where trains have to stop—are the sites of primary concern with respect to seed spillage. Based on the results of the Austrian case study we discuss common measures to limit crop seed spillage which include intensified controls at border railway stations and the mode of seed packing during transportation. We further recommend sufficient cleaning both of goods wagons and of loading areas of trucks and ships as well as an appropriate weed management.

A 4-year record of sitka spruce and western hemlock seed fall on the Cascade Head Experimental Forest.

Science.gov (United States)

Robert H. Ruth; Carl M. Berntsen

1955-01-01

Four years' measurement of seed fall in the spruce-hemlock type on the Cascade Head Experimental Forest indicates that an ample supply of seed is distributed over clear-cut areas under staggered-setting cutting. The largest tract sampled was 81 acres; in spite of a seed crop failure in 1950, it received an average of 243,000 viable spruce and hemlock seeds per...

Pulmonary heart valve replacement using stabilized acellular xenogeneic scaffolds; effects of seeding with autologous stem cells

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Harpa Marius Mihai

2015-12-01

Full Text Available Background: We hypothesized that an ideal heart valve replacement would be acellular valve root scaffolds seeded with autologous stem cells. To test this hypothesis, we prepared porcine acellular pulmonary valves, seeded them with autologous adipose derived stem cells (ADSCs and implanted them in sheep and compared them to acellular valves.

Sliding contact loading enhances the tensile properties of mesenchymal stem cell-seeded hydrogels

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AH Huang

2012-07-01

Full Text Available The primary goal of cartilage tissue engineering is to recapitulate the functional properties and structural features of native articular cartilage. While there has been some success in generating near-native compressive properties, the tensile properties of cell-seeded constructs remain poor, and key features of cartilage, including inhomogeneity and anisotropy, are generally absent in these engineered constructs. Therefore, in an attempt to instill these hallmark properties of cartilage in engineered cell-seeded constructs, we designed and characterized a novel sliding contact bioreactor to recapitulate the mechanical stimuli arising from physiologic joint loading (two contacting cartilage layers. Finite element modeling of this bioreactor system showed that tensile strains were direction-dependent, while both tensile strains and fluid motion were depth-dependent and highest in the region closest to the contact surface. Short-term sliding contact of mesenchymal stem cell (MSC-seeded agarose improved chondrogenic gene expression in a manner dependent on both the axial strain applied and transforming growth factor-β supplementation. Using the optimized loading parameters derived from these short-term studies, long-term sliding contact was applied to MSC-seeded agarose constructs for 21 d. After 21 d, sliding contact significantly improved the tensile properties of MSC-seeded constructs and elicited alterations in type II collagen and proteoglycan accumulation as a function of depth; staining for these matrix molecules showed intense localization in the surface regions. These findings point to the potential of sliding contact to produce engineered cartilage constructs that begin to recapitulate the complex mechanical features of the native tissue.

The direct biologic effects of radioactive 125I seeds on pancreatic cancer cells PANC-1, at continuous low-dose rates.

Science.gov (United States)

Wang, Jidong; Wang, Junjie; Liao, Anyan; Zhuang, Hongqing; Zhao, Yong

2009-08-01

The relative biologic effectiveness of model 6711 125I seeds (Ningbo Junan Pharmaceutical Technology Company,Ningbo, China) and their effects on growth, cell cycle, and apoptosis in human pancreatic cancer cell line PANC-1 were examined in the present study. PANC-1 cells were exposed to the absorbed doses of 1, 2, 4, 6, 8, and 10 Gyeither with 125I seeds (initial dose rate, 2.59 cGy=h) or with 60Co g-ray irradiation (dose rate, 221 cGy=min),respectively. Significantly greater numbers of apoptotic PANC-1 cells were detected following the continuouslow-dose-rate (CLDR) irradiation of 125I seeds, compared with cells irradiated with identical doses of 60Co g-ray. The D(0) for 60Co g-ray and 125I seed irradiation were 2.30 and 1.66, respectively. The survival fraction after 125Iseed irradiation was significantly lower than that of 60Co g-ray, with a relative biologic effectiveness of 1.39.PANC-1 cells were dose dependently arrested in the S-phase by 60Co g-rays and in the G2=M phase by 125I seeds,24 hour after irradiation. CLDR irradiation by 125I seeds was more effective in inducing cell apoptosis in PANC-1cells than acute high-dose-rate 60Co g irradiation. Interestingly, CLDR irradiation by 125I seeds can cause PANC-1cell-cycle arrest at the G2=M phase and induce apoptosis, which may be an important mechanism underlying 125Iseed-induced PANC-1 cell inhibition.

Jatropha curcas seed oil as a viable source for biodiesel

International Nuclear Information System (INIS)

Anwar, F.; Jamil, A.; Bhatti, H.N.; Rashid, U.

2010-01-01

The purpose of the present study was to explore the utility of Jatropha (Jatropha curcas) seed oil for bio diesel production. The preliminarily evaluated Jatropha oil was transmethylated under optimized set of reaction conditions: methanol/oil molar ratio (6:1), sodium methoxide catalyst concentration (1.00%), temperature (65 deg. C) and mixing intensity (600 rpm) providing 94.00% yield of Jatropha oil methyl esters (JOMEs)/biodiesel. The gas chromatographic (GC) analysis showed that JOMEs mainly comprised of six fatty acids: linoleic (49.75%), stearic (16.80%), oleic (13.00%), palmitic (12.15%), arachidic (5.01%) and gadoleic (2.00%) acids. 1H-NMR spectrum of JOMEs was also recorded. The thermal stability of the JOMEs produced was assessed by thermo gravimetric analysis (TGA). The fuel properties of the biodiesel produced were found to be within the standards specifications of ASTM D 6751 and EN 14214. (author)

Effects of foliar boron application on seed composition, cell wall boron, and seed δ15N and δ13C isotopes in water-stressed soybean plants

Science.gov (United States)

Bellaloui, Nacer; Hu, Yanbo; Mengistu, Alemu; Kassem, My A.; Abel, Craig A.

2013-01-01

Limited information is available on the effects of foliar boron (B) application on soybean seed composition. The objective of this research was to investigate the effects of foliar B on seed composition (protein, oil, fatty acids, and sugars). Our hypothesis was that since B is involved in nitrogen and carbon metabolism, it may impact seed composition. A repeated greenhouse experiment was conducted where half of the soybean plants was exposed to water stress (WS) and the other half was well-watered. Foliar boron (FB) in the form of boric acid was applied twice at a rate of 1.1 kg ha−1. The first application was during flowering stage, and the second application was during seed-fill stage. Treatments were water stressed plants with no FB (WS–B); water stressed plants with FB (WS+B); watered plants without FB (W–B), and watered plants with FB (W+B). The treatment W–B was used as a control. Comparing with WS–B plants, B concentration was the highest in leaves and seed of W+B plants (84% increase in leaves and 73% in seed). Seeds of W+B plants had higher protein (11% increase), oleic acid (27% increase), sucrose (up to 40% increase), glucose, and fructose comparing with W–B. However, seed stachyose concentrations increased by 43% in WS–B plants seed compared with W–B plants. Cell wall (structural) B concentration in leaves was higher in all plants under water stress, especially in WS–B plants where the percentage of cell wall B reached up to 90%. Water stress changed seed δ15N and δ13C values in both B applied and non-B applied plants, indicating possible effects on nitrogen and carbon metabolism. This research demonstrated that FB increased B accumulation in leaves and seed, and altered seed composition of well-watered and water stressed plants, indicating a possible involvement of B in seed protein, and oleic and linolenic fatty acids. Further research is needed to explain mechanisms of B involvement in seed protein and fatty acids. PMID:23888163

Response of sunflower to various pre-germination techniques for breaking seed dormancy

International Nuclear Information System (INIS)

Nasreen, S.; Khan, M.A.; Uddin, S.

2015-01-01

Seed dormancy is considered to be a serious constraint in sunflower seed production. Viable seeds sometimes do not germinate even in the presence of favorable environmental conditions. Such seeds are suspected to be dormant. The study was conducted under controlled/laboratory conditions during spring 2010 at National Agricultural Research Centre, Islamabad. The objective of the study was to evaluate some techniques to convert a seed from dormant to non-dormant germinable state. Dormant seeds of 21 sunflower hybrids were treated with three hot water treatments (100/80 degree C) and four chemicals potassium nitrate, 0.2%, thiourea, 0.5%, ethanol, 25%, acetone, 25% for breaking seed dormancy .The untreated seed was taken as control. Soaking seeds in hot water (80 degree C) for 15 minutes followed by one day dry and seed treatment with acetone were found to be the most effective and successful techniques in converting the seed from dormant to non-dormant state. (author)

DISE: A Seed-Dependent RNAi Off-Target Effect That Kills Cancer Cells.

Science.gov (United States)

Putzbach, William; Gao, Quan Q; Patel, Monal; Haluck-Kangas, Ashley; Murmann, Andrea E; Peter, Marcus E

2018-01-01

Off-target effects (OTEs) represent a significant caveat for RNAi caused by substantial complementarity between siRNAs and unintended mRNAs. We now discuss the existence of three types of seed-dependent OTEs (sOTEs). Type I involves unintended targeting through the guide strand seed of an siRNA. Type II is caused by the activity of the seed on the designated siRNA passenger strand when loaded into the RNA-induced silencing complex (RISC). Both type I and II sOTEs will elicit unpredictable cellular responses. By contrast, in sOTE type III the guide strand seed preferentially targets essential survival genes resulting in death induced by survival gene elimination (DISE). In this Opinion article, we discuss DISE as a consequence of RNAi that may preferentially affect cancer cells. Copyright © 2017 Elsevier Inc. All rights reserved.

DISPERSAL OF SEEDS AS NEST MATERIAL BY THE CACTUS WREN

Science.gov (United States)

Cactus wren (Campylorhynchus brunneicapillus) nests from the southern Chihuahuan Desert contained viable seeds of grasses, forbs, and shrubs. The most common plants used as construction material in these nests were Muhlenbergia porteri, Boerhavia spicata, and the alien grass Era...

Behavior of Jatropha curcas L. seeds under osmotic stress: germination and cell cycle activity

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Cristiane Dantas de Brito

2015-08-01

Full Text Available Jatropha curcas is an oil-rich Euphorbiaceae seed species renowned for its apparent tolerance to environmental stresses. It is considered a promising source of renewable feedstock for biodiesel production in the Brazilian semiarid region where crop establishment requires a better understanding of the mechanisms leading to proper seed and plant behavior under water restrictive conditions. This study describes physiological and cytological profiles of J. curcas seeds imbibed in water restriction conditions by means of osmotic stress or osmoconditioning. Seeds were characterized by size, weight, moisture content and dry mass, germinability, and cell cycle activation by means of tubulin and microtubule cytoskeleton accumulation. Osmoconditioning at -0.8 MPa did not induce priming effects as it did not improve the physiological quality of the seed lots. Western blotting and immunocytochemical analysis revealed an increasing accumulation of tubulin and microtubule cytoskeleton in seeds imbibed in water for 48h onwards, culminating in the onset of mitotic configurations after germination. Only cortical microtubules were observed during seed osmoconditioning, whereas mitotic microtubules only occurred after re-imbibition of osmoconditioned seeds in water and subsequent germination.

Single-Kernel FT-NIR Spectroscopy for Detecting Supersweet Corn (Zea mays L. Saccharata Sturt Seed Viability with Multivariate Data Analysis

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Guangjun Qiu

2018-03-01

Full Text Available The viability and vigor of crop seeds are crucial indicators for evaluating seed quality, and high-quality seeds can increase agricultural yield. The conventional methods for assessing seed viability are time consuming, destructive, and labor intensive. Therefore, a rapid and nondestructive technique for testing seed viability has great potential benefits for agriculture. In this study, single-kernel Fourier transform near-infrared (FT-NIR spectroscopy with a wavelength range of 1000–2500 nm was used to distinguish viable and nonviable supersweet corn seeds. Various preprocessing algorithms coupled with partial least squares discriminant analysis (PLS-DA were implemented to test the performance of classification models. The FT-NIR spectroscopy technique successfully differentiated viable seeds from seeds that were nonviable due to overheating or artificial aging. Correct classification rates for both heat-damaged kernels and artificially aged kernels reached 98.0%. The comprehensive model could also attain an accuracy of 98.7% when combining heat-damaged samples and artificially aged samples into one category. Overall, the FT-NIR technique with multivariate data analysis methods showed great potential capacity in rapidly and nondestructively detecting seed viability in supersweet corn.

Single-Kernel FT-NIR Spectroscopy for Detecting Supersweet Corn (Zea mays L. Saccharata Sturt) Seed Viability with Multivariate Data Analysis.

Science.gov (United States)

Qiu, Guangjun; Lü, Enli; Lu, Huazhong; Xu, Sai; Zeng, Fanguo; Shui, Qin

2018-03-28

The viability and vigor of crop seeds are crucial indicators for evaluating seed quality, and high-quality seeds can increase agricultural yield. The conventional methods for assessing seed viability are time consuming, destructive, and labor intensive. Therefore, a rapid and nondestructive technique for testing seed viability has great potential benefits for agriculture. In this study, single-kernel Fourier transform near-infrared (FT-NIR) spectroscopy with a wavelength range of 1000-2500 nm was used to distinguish viable and nonviable supersweet corn seeds. Various preprocessing algorithms coupled with partial least squares discriminant analysis (PLS-DA) were implemented to test the performance of classification models. The FT-NIR spectroscopy technique successfully differentiated viable seeds from seeds that were nonviable due to overheating or artificial aging. Correct classification rates for both heat-damaged kernels and artificially aged kernels reached 98.0%. The comprehensive model could also attain an accuracy of 98.7% when combining heat-damaged samples and artificially aged samples into one category. Overall, the FT-NIR technique with multivariate data analysis methods showed great potential capacity in rapidly and nondestructively detecting seed viability in supersweet corn.

Separation of viable lactic acid bacteria from fermented milk

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Tomohiko Nishino

2018-04-01

Full Text Available Probiotics are live microorganisms that provide health benefits to humans. Some lactic acid bacteria (LAB are probiotic organisms used in the production of fermented foods, such as yogurt, cheese, and pickles. Given their widespread consumption, it is important to understand the physiological state of LAB in foods such as yogurt. However, this analysis is complicated, as it is difficult to separate the LAB from milk components such as solid curds, which prevent cell separation by dilution or centrifugation. In this study, we successfully separated viable LAB from yogurt by density gradient centrifugation. The recovery rate was >90 %, and separation was performed until the stationary phase. Recovered cells were observable by microscopy, meaning that morphological changes and cell viability could be directly detected at the single-cell level. The results indicate that viable LAB can be easily purified from fermented milk. We expect that this method will be a useful tool for the analysis of various aspects of probiotic cells, including their enzyme activity and protein expression. Keywords: Food analysis, Microbiology

Schwann cell seeded guidance tubes restore erectile function after ablation of cavernous nerves in rats.

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May, F; Weidner, N; Matiasek, K; Caspers, C; Mrva, T; Vroemen, M; Henke, J; Lehmer, A; Schwaibold, H; Erhardt, W; Gänsbacher, B; Hartung, R

2004-07-01

Dissection of the cavernous nerves eliminates spontaneous erections. We evaluated the ability of Schwann cell seeded nerve guidance tubes to restore erections after bilateral cavernous nerve resection in rats. Sections (5 mm) of the cavernous nerve were excised bilaterally, followed by immediate bilateral microsurgical reconstruction. In 10 animals per group (20 study nerves) reconstruction was performed by genitofemoral nerve interposition, interposition of silicone tubes or interposition of silicone tubes seeded with homologous Schwann cells. As the control 10 animals (20 study nerves) underwent sham operation (positive control) and bilateral nerve ablation (without reconstruction) was performed in a further 10 (negative control). Erectile function was evaluated 3 months postoperatively by relaparotomy, electrical nerve stimulation and intracavernous pressure recording. After 3 months neurostimulation resulted in an intact erectile response in 90% (18 of 20) of Schwann cell grafts, while treatment with autologous nerves (30% or 6 of 20) or tubes only (50% or 10 of 20) was less successful (p Schwann cell grafts compared to results in the other treatment groups (p Schwann cell grafts. Schwann cell seeded guidance tubes restore erectile function after the ablation of cavernous nerves in rats and they are superior to autologous nerve grafts.

Effects of cell-to-collagen ratio in stem cell-seeded constructs for Achilles tendon repair.

Science.gov (United States)

Juncosa-Melvin, Natalia; Boivin, Gregory P; Galloway, Marc T; Gooch, Cindi; West, John R; Butler, David L

2006-04-01

The objective of the present study was to test the hypotheses that implantation of cell-seeded constructs in a rabbit Achilles tendon defect model would 1) improve repair biomechanics and matrix organization and 2) result in higher failure forces than measured in vivo forces in normal rabbit Achilles tendon (AT) during an inclined hopping activity. Autogenous tissue-engineered constructs were fabricated in culture between posts in the wells of silicone dishes at four cell-to-collagen ratios by seeding mesenchymal stem cells (MSC) from 18 adult rabbits at each of two seeding densities (0.1 x 10(6) and 1 x 10(6) cell/mL) in each of two collagen concentrations (1.3 and 2.6 mg/mL). After 5 days of contraction, constructs having the two highest ratios (0.4 and 0.8 M/mg) were damaged by excessive cell traction forces and could not be used in subsequent in vivo studies. Constructs at the lower ratios (0.04 and 0.08 M/mg) were implanted in bilateral, 2 cm long gap defects in the rabbit's lateral Achilles tendon. At 12 weeks after surgery, both repair tissues were isolated and either failed in tension (n = 13) to determine their biomechanical properties or submitted for histological analysis (n = 5). No significant differences were observed in any structural or mechanical properties or in histological appearance between the two repair conditions. However, the average maximum force and maximum stress of these repairs achieved 50 and 85% of corresponding values for the normal AT and exceeded the largest peak in vivo forces (19% of failure) previously recorded in the rabbit AT. Average stiffness and modulus were 60 and 85% of normal values, respectively. New constructs with lower cell densities and higher scaffold stiffness that do not excessively contract and tear in culture and that further improve the repair stiffness needed to withstand various levels of expected in vivo loading are currently being investigated.

Effects of radioactive 125I seeds on A549 cell line and human embryonic lung diploid cell line 2BS cultivated in vitro and assessment of its clinical safety dose

International Nuclear Information System (INIS)

Bian Wenchao; Qi Liangchen

2012-01-01

Objective: To observe the cell count changes of A549 cell line and human embryonic lung diploid cell line 2BS after irradiated by 125 I seeds with different doses, and to study the growth inhibition of 125 I on this two kinds of cell lines, and to determine its clinical safety dose in treatment of non-small cell lung. Methods: 125 I seeds with different doses (low dose: 0.2 mCi, mediate dose: 0.4 mCi, high dose: 0.8 mCi) were chosen and put into A549 cells and human embryonic lung diploid cell line 2BS in vitro, the cells on the 2nd, 4th, 6th and 8th days after irradiation were collected, the alive cells were counted by cells dyeing experiments, then the growth curves were drawn, and the IC 50 of the radioactive 125 I seeds to both two cell lines were calculated. Results: Compared with blank and control groups, the cell proliferation trend of A549 cells in low dose group was not significantly influenced (P>0.05), but the growth of A549 cells in mediate and high dose groups were inhibited in a time-dependent manner, there were significant differences (P<0.05), the most obvious change was on the 6th day. The IC 50 of the radioactive 125 I seeds to A549 cells was about .04 mCi. While the growth inhibition of 125 I 2BS had no statistically significant differences between various dose groups (P>0.05), and the IC 50 of the radioactive 125 I seeds to 2BS cell line was about 1.65 mCi. Conclusion: 0.4 mCi of radioactive 125 I seeds has already had the obvious damage effect on A549 cell, 0.8 mCi of radioactive 125 I seeds has the stronger effect. The IC 50 of the radioactive 125 I seeds to 2BS cells is about 1.65 mCi, so the clinical safety dosage is 0.4-0.8 mCi. (authors)

On-Chip Dielectrophoretic Separation and Concentration of Viable, Non-Viable and Viable but Not Culturable (VBNC) Escherichia coli

Energy Technology Data Exchange (ETDEWEB)

Packard, M M; Shusteff, M; Alocilja, E C

2012-04-12

Although bacterial culture remains the gold standard for detection of viable bacteria in environmental specimens, the typical time requirement of twenty-four hours can delay and even jeopardize appropriate public health intervention. In addition, culture is incapable of detecting viable but not culturable (VBNC) species. Conversely, nucleic acid and antibody-based methods greatly decrease time to detection but rarely characterize viability of the bacteria detected. Through selection by membrane permeability, the method described in this work employs positive dielectrophoresis (pDEP) for separation and purification of viable and VBNC species from water and allows concentration of bacteria for downstream applications.

Collagen-coated polylactic-glycolic acid (PLGA) seeded with neural-differentiated human mesenchymal stem cells as a potential nerve conduit.

Science.gov (United States)

Sulong, Ahmad Fadzli; Hassan, Nur Hidayah; Hwei, Ng Min; Lokanathan, Yogeswaran; Naicker, Amaramalar Selvi; Abdullah, Shalimar; Yusof, Mohd Reusmaazran; Htwe, Ohnmar; Idrus, Ruszymah Bt Hj; Haflah, Nor Hazla Mohamed

2014-01-01

Autologous nerve grafts to bridge nerve gaps pose various drawbacks. Nerve tissue engineering to promote nerve regeneration using artificial neural conduits has emerged as a promising alternative. To develop an artificial nerve conduit using collagen-coated polylactic-glycolic acid (PLGA) and to analyse the survivability and propagating ability of the neuro-differentiated human mesenchymal stem cells in this conduit. The PLGA conduit was constructed by dip-molding method and coated with collagen by immersing the conduit in collagen bath. The ultra structure of the conduits were examined before they were seeded with neural-differentiated human mesenchymal stem cells (nMSC) and implanted sub-muscularly on nude mice thighs. The non-collagen-coated PLGA conduit seeded with nMSC and non-seeded non-collagen-coated PLGA conduit were also implanted for comparison purposes. The survivability and propagation ability of nMSC was studied by histological and immunohistochemical analysis. The collagen-coated conduits had a smooth inner wall and a highly porous outer wall. Conduits coated with collagen and seeded with nMSCs produced the most number of cells after 3 weeks. The best conduit based on the number of cells contained within it after 3 weeks was the collagen-coated PLGA conduit seeded with neuro-transdifferentiated cells. The collagen-coated PLGA conduit found to be suitable for attachment, survival and proliferation of the nMSC. Minimal cell infiltration was found in the implanted conduits where nearly all of the cells found in the cell seeded conduits are non-mouse origin and have neural cell markers, which exhibit the biocompatibility of the conduits. The collagen-coated PLGA conduit is biocompatible, non-cytotoxic and suitable for use as artificial nerve conduits.

Conditions Affecting Shelf-Life of Inoculated Legume Seed

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Greg Gemell

2012-02-01

Full Text Available Microbial inoculants are becoming more available as sustainable alternatives to fertilizers and other agrichemicals in broad-acre cropping. However, with the exception of legume inoculants little is understood about effective delivery and survival of the inoculum. Legume inoculants are applied to both seed and soil but seed inoculation is the most economical technique. Large quantities of pasture seed in Australia are inoculated by commercial seed coating companies, but the long-term survival of seed-applied inoculum is variable and monitoring of viability requires specialist microbiology skills and facilities. The aim of our research was to define optimum storage conditions for survival of rhizobia on legume seed and evaluate water activity as a means of monitoring shelf-life. The relationship between survival and water activity varied according to seed species, inoculum preparation, coating ingredients, initial water activity and time suggesting that storage conditions would need to be defined for each different combination. Although drying seeds after coating significantly reduced viable numbers of rhizobia, survival of rhizobia on dried commercially coated lucerne seed after 11 weeks was less variable than seeds that had not been dried. The highest numbers were maintained when seeds remained dry with water activities of between 0.47 and 0.38. The quality of inoculated seed could be improved by reducing the death rate of inoculum during preparation and providing optimum storage conditions for long-term survival.

Cytological changes of root tip cells of alfalfa seeds after space flight

International Nuclear Information System (INIS)

Ren Weibo; Xu Zhu; Chen Libo; Guo Huiqin; Wang Mi; Zhao Liang

2008-01-01

To understand the cytological effects of space flight on alfalfa seeds, dry seeds of three lines (Line 1, Line 2 and Line 4) were selected and loaded onto 'Shijian No.8' satellite for space flight. After returning to the ground, root tips of alfalfa were clipped and chromosome aberrations were observed by microscope. Data showed that space flight had two types of effect on cell mitotic: one was positive (Line 2, Line 4) and the other was negative (Line 1). Such chromosome aberrations were observed as micronucleus, chromosome bridge, fragments, lagging and so on. The frequency of aberration varied with the different materials. Conclusion was that space flight had significant effect on root tip cells, which mainly showed as the chromosome aberrations. (authors)

Human umbilical cord mesenchymal stem cells: osteogenesis in vivo as seed cells for bone tissue engineering.

Science.gov (United States)

Diao, Yinze; Ma, Qingjun; Cui, Fuzhai; Zhong, Yanfeng

2009-10-01

Mesenchymal stem cells (MSCs) are ideal seed cells for bone tissue engineering. However, intrinsic deficiencies exist for the autologous transplantation strategy of constructing artificial bone with MSCs derived from bone marrow of patients. In this study, MSCs-like cells were isolated from human umbilical cords and were expanded in vitro. Flow cytometric analysis revealed that cells from the fourth passage were positive for CD29, CD44, CD71, CD73, CD90, and CD105 whereas they were negative for CD14, CD34, CD45, and CD117. Furthermore, these cells expressed HLA-A, B, C (MHC-I), but not HLA-DP, DQ, DR (MHC-II), or costimulatory molecules such as CD80 and CD86. Following incubation in specific inductive media for 3 weeks, cultured cells were shown to possess potential to differentiate into adipogenic, osteogenic or chondrogenic lineages in vitro. The umbilical cord-derived MSCs (UC-MSCs) were loaded with a biomimetic artificial bone scaffold material before being implanted subcutaneously in the back of Balb/c nude mice for four to twelve weeks. Our results revealed that UC-MSCs loaded with the scaffold displayed capacity of osteogenic differentiation leading to osteogenesis with human origin in vivo. As a readily available source of seed cells for bone tissue engineering, UC-MSCs should have broad application prospects.

Generation of viable progeny from dead brooders of endangered catfish Clarias magur (Hamilton, 1822

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Sullip Kumar Majhi

2017-10-01

Full Text Available The obligatory air-breathing catfish Clarias magur is a prime candidate for aquaculture owing to its unique taste, high growth rate, and hardy nature. However, recently the IUCN has listed the species under the endangered category because the population has critically declined in the wild. The sexually mature C. magur brooders are often collected from their natural habitats for seed production in captivity. In many cases, the brooder dies due to handling injuries or confinement stress. In this study, we demonstrated that viable progeny could be generated from freshly dead sexually mature C. magur. Three hours after death, the gonads were excised, macroscopically examined and gamete viability was evaluated. Artificial fertilization was performed by mixing the sperm suspension with the eggs. Water was added after 1 min of mixing to activate the fertilization process. We observed 85%-93% fertilization success from gametes derived from dead donors as opposed to 90%-95% from those derived from live control donors. The embryos showed normal development and resulted in the generation of 88%-92% viable progeny, which was similar to the progeny derived from control donors (92%-93%. The results obtained in this study will have profound implications in enhancing the seed production of endangered C. magur and could potentially be applied to other key commercially or endangered fish species. Keywords: Biological sciences, Developmental biology, Zoology

Wavelet-SVM classification and automatic recognition of unstained viable cells in phase-contrast microscopy

International Nuclear Information System (INIS)

Skoczylas, M.; Rakowski, W.; Cherubini, R.; Gerardi, S.

2011-01-01

Irradiation of individual cultured mammalian cells with a pre-selected number of ions down to one ion per single cell is a useful experimental approach to investigating the low-dose ionising radiation exposure effects and thus contributing to a more realistic human cancer risk assessment. One of the crucial tasks of all the microbeam apparatuses is the visualisation, recognition and positioning of every individual cell of the cell culture to be irradiated. Before irradiations, mammalian cells (specifically, Chinese hamster V79 cells) are seeded and grown as a monolayer on a mylar surface used as the bottom of a specially designed holder. Manual recognition of unstained cells in a bright-field microscope is a time-consuming procedure; therefore, a parallel algorithm has been conceived and developed in order to speed up this irradiation protocol step. Many technical problems have been faced to overcome the complexity of the images to be analysed: cell discrimination in an inhomogeneous background, among many disturbing bodies mainly due to the mylar surface roughness and culture medium bodies; cell shapes, depending on how they attach to the surface, which phase of the cell cycle they are in and on cell density. Preliminary results of the recognition and classification based on a method of wavelet kernels for the support vector machine classifier will be presented. (authors)

Cell signaling mechanisms and metabolic regulation of germination and dormancy in barley seeds

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Zhenguo Ma

2017-12-01

Full Text Available During germination of barley (Hordeum vulgare L. seeds, important morphological and physiological changes take place, including development of organs and tissues and activation of metabolic pathways. Germination and dormancy of seeds are regulated by abscisic acid, gibberellins, reactive oxygen species (ROS, reactive nitrogen species (RNS and several other factors. Activities of ascorbate–glutathione cycle enzymes, responsible for scavenging ROS, strongly increase. Catalase and superoxide dismutase activities, also scavenging ROS, decrease at the onset of seed germination and then increase. With the increase in aerobic metabolism after radicle protrusion, the activities of the fermentation enzymes lactate and alcohol dehydrogenase decline rapidly. The RNS-scavenging activity of S-nitrosoglutathione reductase decreases in the course of seed germination, in concert with elevation of nitric oxide production and protein nitrosylation. This activity supports the role of RNS in regulating seed germination. Transcription of various genes at different phases of seed germination exhibits phase-specific changes. During imbibition, genes involved in cell wall metabolism are highly expressed; in the middle phase of seed germination before radicle protrusion, genes involved in amino acid synthesis, protein synthesis, and transport and nucleic acid synthesis are upregulated significantly, and after radicle protrusion, genes involved in photosynthetic metabolism are induced. In summary, signal transduction and metabolic regulation of seed germination involve diverse reactions and complex regulation at different levels of metabolic organization. Keywords: Seed germination, Reactive oxygen species, Reactive nitrogen species, Signal transduction, Gene expression

Endothelial cell seeding on crosslinked collagen : Effects of crosslinking on endothelial cell proliferation and functional parameters

NARCIS (Netherlands)

Wissink, MJB; van Luyn, MJA; Dijk, F; Poot, AA; Engbers, GHM; Beugeling, T; van Aken, WG; Feijen, J

Endothelial cell seeding, a promising method to improve the performance of small-diameter vascular grafts, requires a suitable substrate, such as crosslinked collagen. Commonly used crosslinking agents such as glutaraldehyde and formaldehyde cause, however, cytotoxic reactions and thereby hamper

Characterization of the Viable but Nonculturable (VBNC State in Saccharomyces cerevisiae.

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Mohammad Salma

Full Text Available The Viable But Non Culturable (VBNC state has been thoroughly studied in bacteria. In contrast, it has received much less attention in other microorganisms. However, it has been suggested that various yeast species occurring in wine may enter in VBNC following sulfite stress.In order to provide conclusive evidences for the existence of a VBNC state in yeast, the ability of Saccharomyces cerevisiae to enter into a VBNC state by applying sulfite stress was investigated. Viable populations were monitored by flow cytometry while culturable populations were followed by plating on culture medium. Twenty-four hours after the application of the stress, the comparison between the culturable population and the viable population demonstrated the presence of viable cells that were non culturable. In addition, removal of the stress by increasing the pH of the medium at different time intervals into the VBNC state allowed the VBNC S. cerevisiae cells to "resuscitate". The similarity between the cell cycle profiles of VBNC cells and cells exiting the VBNC state together with the generation rate of cells exiting VBNC state demonstrated the absence of cellular multiplication during the exit from the VBNC state. This provides evidence of a true VBNC state. To get further insight into the molecular mechanism pertaining to the VBNC state, we studied the involvement of the SSU1 gene, encoding a sulfite pump in S. cerevisiae. The physiological behavior of wild-type S. cerevisiae was compared to those of a recombinant strain overexpressing SSU1 and null Δssu1 mutant. Our results demonstrated that the SSU1 gene is only implicated in the first stages of sulfite resistance but not per se in the VBNC phenotype. Our study clearly demonstrated the existence of an SO2-induced VBNC state in S. cerevisiae and that the stress removal allows the "resuscitation" of VBNC cells during the VBNC state.

Evaluation of early healing events around mesenchymal stem cell-seeded collagen-glycosaminoglycan scaffold. An experimental study in Wistar rats.

LENUS (Irish Health Repository)

Alhag, Mohamed

2011-03-01

Tissue engineering using cell-seeded biodegradable scaffolds offers a new bone regenerative approach that might circumvent many of the limitations of current therapeutic modalities. The aim of this experiment was to study the early healing events around mesenchymal stem cell-seeded collagen-glycosaminoglycan scaffolds.

Depth of dormancy in tomato seeds is related to the progression of the cell cycle prior to its induction

NARCIS (Netherlands)

Castro, de R.D.; Lammeren, van A.A.M.; Groot, S.P.C.; Bino, R.J.; Hilhorst, H.W.M.

2001-01-01

Cell cycle activities are initiated following imbibition of non-dormant seeds. However, it is not known whether cell cycle related events other than DNA replication also remain suppressed in imbibed dormant seeds. The objective of this study was to demonstrate that the transitions between the

Nano-Micelle of Moringa Oleifera Seed Oil Triggers Mitochondrial Cancer Cell Apoptosis

Science.gov (United States)

Abd-Rabou, Ahmed A; Zoheir, Khairy M A; Kishta, Mohamed S; Shalby, Aziza B; Ezzo, Mohamed I

2016-01-01

Cancer, a worldwide epidemic disease with diverse origins, involves abnormal cell growth with the potential to invade other parts of the body. Globally, it is the main cause of mortality and morbidity. To overcome the drawbacks of the commercially available chemotherapies, natural products-loaded nano-composites are recommended to improve cancer targetability and decrease the harmful impact on normal cells. This study aimed at exploring the anti-cancer impacts of Moringa oleifera seed oil in its free- (MO) and nano-formulations (MOn) through studying whether it mechanistically promotes mitochondrial apoptosis-mediating cell death. Mitochondrial-based cytotoxicity and flow cytometric-based apoptosis analyses were performed on cancer HepG2, MCF7, HCT 116, and Caco-2 cell lines against normal kidney BHK-21 cell line. The present study resulted that MOn triggered colorectal cancer Caco-2 and HCT 116 cytotoxicity via mitochondrial dysfunction more powerful than its free counterpart (MO). On the other side, MOn and MO remarkably induces HCT 116 mitochondrial apoptosis, while sparing normal BHK-21 cells with minimal cytotoxic effect. The present results concluded that nano-micelle of Moringa oleifera seed oil (MOn) can provide a novel therapeutic approach for colorectal and breast cancers via mitochondrial-mediated apoptosis, while sparing normal and even liver cancer cells a bit healthy or with minimal harmful effect. Intriguingly, MOn induced breast cancer not hepatocellular carcinoma cell death. PMID:28032498

Bystander effects of exposure to low-dose-rate 125I seeds on human lung cancers cells in vitro

International Nuclear Information System (INIS)

Jia Rongfei; Chen Honghong; Yu Lei; Zhao Meijia; Shao Chunlin; Cheng Wenying

2007-01-01

The bystander effects induced by continuous low-dose-rate (LDR) 125 I seeds radiation on damage of human lung cancer cells were investigated. Human adenocarcinoma cell line A549 and human small cell lung cancer cell line NCI-H446, which have different sensitivities to high-dose rate (HDR) external irradiation, were exposed directly to 125 I seeds in vitro and co-cultured with unirradiated cells for 24 h. Using cytokinesis-blocking micronucleus method and γ H2AX fluorescence immunoassay, bystander effects induced by 2Gy and 4Gy 125 I seed irradiation on micronucleus formation and DNA double-strand breaks (DSBs) of human lung cancer cells were detected and evaluated. The results showed that irradiation with 125 I seeds can induce medium-mediated bystander effects in A549 cells and NCI-H446 cells, exhibiting that both micronuclei formation and γ H2AX focus formation in bystander cells were increased significantly compared with non-irradiated cells. The extent of DNA damage induced by bystander effects was correlated with accumulated radiation dose and radiosensitive of tumor cells. NCI-H446 cells that were sensitive to HDR γ irradiation were more sensitive to continuous LDR irradiation and bystander effects than A549. However, a comparison between the bystander effects and direct effects elicits the intensity of bystander responses of A549 cells was higher than that of NCI-H446 cells. A dose-related reduction in bystander responses was observed both in A549 cells and NCI-H446 cells, suggesting that the signaling factors involved in the bystander signaling pathways may decrease with the increase of cell damages. (authors)

Deleterious effects on MDAMB-231 breast adenocarcinoma cell lineage submitted to Ho-166 radioactive seeds at very low activity

Energy Technology Data Exchange (ETDEWEB)

Falcao, Patricia L.; Campos, Tarcisio P.R., E-mail: campos@nuclear.ufmg.br [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Dept. de Engenharia Nuclear; Sarmento, Eduardo V. [Centro de Desenvolvimento de Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil); Cuperschmid, Ethel M. [Universidade Federal de Minas Gerais (CEMEMOR/UFMG), Belo Horizonte, BR (Brazil). Fac. de Medicina. Centro de Memoria da Medicina

2011-07-01

Herein, the deleterious effect of ionizing radiation provided by Ho-166 radioactive seeds at low activity were addressed, based on experimental in vitro assays at the MDA MB231 cell lineage, a breast adenocarcinoma, compared to PBMC - peripheral blood cells. The methodology involves of the MDBMB-231 and PBMC expansion in culture in suitable environment in 30mm well plates and T-25 flasks. Seeds were synthesized with Ho-165 incorporated and characterized previously. Activation was processed at IPR1 reactor at the peripheral table, at 8h exposition. Three groups of seeds were tested: 0,34 mCi, 0,12 mCi activity, and control group. Such seeds were placed on culture and held to a period of 05 half-lives of the radionuclide. The biological responses at these exposure were documented by inverse microscopic photographic in time. Also, MTT essay were performed. A fast response in producing deleterious effects at cancer cell was observed even if for the low activity seeds. Also, a biological response dependent to a radial distance of the seed was observed. At conclusion, viability clonogenic control of MDAMB231 is identified at the exposition to Ho-166 ceramic seeds, even if at low activity of 0,1 to 0,3mCi. (author)

Deleterious effects on MDAMB-231 breast adenocarcinoma cell lineage submitted to Ho-166 radioactive seeds at very low activity

International Nuclear Information System (INIS)

Falcao, Patricia L.; Campos, Tarcisio P.R.; Cuperschmid, Ethel M.

2011-01-01

Herein, the deleterious effect of ionizing radiation provided by Ho-166 radioactive seeds at low activity were addressed, based on experimental in vitro assays at the MDA MB231 cell lineage, a breast adenocarcinoma, compared to PBMC - peripheral blood cells. The methodology involves of the MDBMB-231 and PBMC expansion in culture in suitable environment in 30mm well plates and T-25 flasks. Seeds were synthesized with Ho-165 incorporated and characterized previously. Activation was processed at IPR1 reactor at the peripheral table, at 8h exposition. Three groups of seeds were tested: 0,34 mCi, 0,12 mCi activity, and control group. Such seeds were placed on culture and held to a period of 05 half-lives of the radionuclide. The biological responses at these exposure were documented by inverse microscopic photographic in time. Also, MTT essay were performed. A fast response in producing deleterious effects at cancer cell was observed even if for the low activity seeds. Also, a biological response dependent to a radial distance of the seed was observed. At conclusion, viability clonogenic control of MDAMB231 is identified at the exposition to Ho-166 ceramic seeds, even if at low activity of 0,1 to 0,3mCi. (author)

Pseudomonas sp. LMTK32 production in modified media for pelleting seeds of maca (Lepidium meyenii Walp.

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Martin Guerra Lopez

2018-05-01

Full Text Available Biomass of Pseudomonas sp. LMTK32 was produced from modification of culture media Yeast Extract Mannitol Broth (YEMB with the aim of increasing the number of viable cells with the ability to promote the germination of maca seeds pelleted with the bacteria and reduce production costs. In the optimization process, the yeast extract and mannitol components of the LMC culture media were replaced by commercial sources of sucrose and glutamate, whose concentrations were determined in flasks by statistical design from Box-Behnken; in addition, the effect of the inoculum percentage on the time of biomass production was determined. Subsequently, it was determined at the bioreactor level that 28.57 h-1 was the adequate value of the volumetric oxygen transfer coefficient (kLa at 600 rpm, producing 1.28 x 10 11 CFU / mL. In the LMC M1 modified media, using 12.06 g / L-1 of sucrose, 11.50 g / L-1 of sodium glutamate and 10.9% of inoculant obtained 15x108 CFU / mL, increasing in 48% the number of viable cells with respect to the YEMB control treatment (7.8x10 8 CFU / mL. At the in vitro level, the pelleting of maca seeds with Pseudomonas sp. LMTK32 produced in bioreactor and in the modified media M1 favored its germination. From commercial organic substrates, efficient bacterial inoculants can be produced in the development of maca crops, without altering their ability to promote plant growth.

Entry of Vibrio harveyi and Vibrio fischeri into the viable but nonculturable state.

Science.gov (United States)

Ramaiah, N; Ravel, J; Straube, W L; Hill, R T; Colwell, R R

2002-01-01

Physiological responses of marine luminous bacteria, Vibrio harveyi (ATCC 14216) and V. fischeri (UM1373) to nutrient-limited normal strength (35 ppt iso-osmolarity) and low (10 ppt hypo-osmolarity) salinity conditions were determined. Plate counts, direct viable counts, actively respiring cell counts, nucleoid-containing cell counts, and total counts were determined. Vibrio harveyi incubated at 22 degrees C in nutrient-limited artificial seawater (ASW) became nonculturable after approximately 62 and 45 d in microcosms of 35 ppt and 10 ppt ASW, respectively. In contrast, V. fischeri became nonculturable at approximately 55 and 31 d in similar microcosms. Recovery of both culturability and luminescence of cells in the viable but nonculturable state was achieved by addition of nutrient broth or nutrient broth supplemented with a carbon source, including luminescence-stimulating compounds. Temperature upshift from 22 degrees C to 30 degrees C or 37 degrees C did not result in recovery from nonculturability. The study confirms entry of V. harveyi and V. fischeri into the viable but nonculturable state under low-nutrient conditions and demonstrates nutrient-dependent resuscitation from this state. This study confirms loss of luminescence of V. harveyi and V. fischeri on entry into the viable but nonculturable state and suggests that enumeration of luminescent cells in water samples may be a rapid method to deduce the nutrient status of a water sample.

Introduce of Viable But Nonculturable Bacteria

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Mehdi Hassanshahian

2008-03-01

Full Text Available Viable-But-Nonculturable-State (VBNC is the condition in which bacteria fail to grow on their routine bacteriological media where they would normally grow and develop into colonies, but are still alive and capable of renewed metabolic activity. VBNC state is useful for evaluating public health and for ascertaining the sterility of drinking water, pharmaceuticals, and foodstuff. A number of bacteria, mostly pathogenic to humans, have been proved to enter into this state in response to natural stresses such as starvation, incubation out of optimum growth temperature, increased osmotic pressure, etc. Once in the VBNC state, they undergo various physiological, structural, and genetic alterations. These alterations result in reduced cell size, conversion from bacilli to coccid, thickened cell walls, and peptidoglycan gaining many cross links. Metabolic changes also occur that include reductions in growth, nutrient transport, and respiratory rate; biosynthesis of new protein, and ATP remaining at a constant level. It has been shown that in the VBNC state, some pathogens conserve their virulence properties. Gene expression continues in the VBNC cell. Nucleic acids remain intact in the early VBNC phase but they gradually undergo degradation with prolonged VBNC. Cytological methods such as direct viable count and reduction of tetrazolium salts, and molecular methods such as reverse transcription polymerase chain reaction and green fluorescent protein have been used for the study of VBNC. Resuscitation from VBNC state starts when the inducing factor(s is/are lifted. Factors that help the resuscitation of VBNC bacteria include addition of certain nutrients and chemicals, introduction of a few culturable cells into the VBNC cell population, and passage through the animal host. As virulence properties are sustained during the VBNC phase, special care must be paid when evaluating sterility of drinking water.

Accelerated craniofacial bone regeneration through dense collagen gel scaffolds seeded with dental pulp stem cells

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Chamieh, Frédéric; Collignon, Anne-Margaux; Coyac, Benjamin R.; Lesieur, Julie; Ribes, Sandy; Sadoine, Jérémy; Llorens, Annie; Nicoletti, Antonino; Letourneur, Didier; Colombier, Marie-Laure; Nazhat, Showan N.; Bouchard, Philippe; Chaussain, Catherine; Rochefort, Gael Y.

2016-12-01

Therapies using mesenchymal stem cell (MSC) seeded scaffolds may be applicable to various fields of regenerative medicine, including craniomaxillofacial surgery. Plastic compression of collagen scaffolds seeded with MSC has been shown to enhance the osteogenic differentiation of MSC as it increases the collagen fibrillary density. The aim of the present study was to evaluate the osteogenic effects of dense collagen gel scaffolds seeded with mesenchymal dental pulp stem cells (DPSC) on bone regeneration in a rat critical-size calvarial defect model. Two symmetrical full-thickness defects were created (5 mm diameter) and filled with either a rat DPSC-containing dense collagen gel scaffold (n = 15), or an acellular scaffold (n = 15). Animals were imaged in vivo by microcomputer tomography (Micro-CT) once a week during 5 weeks, whereas some animals were sacrificed each week for histology and histomorphometry analysis. Bone mineral density and bone micro-architectural parameters were significantly increased when DPSC-seeded scaffolds were used. Histological and histomorphometrical data also revealed significant increases in fibrous connective and mineralized tissue volume when DPSC-seeded scaffolds were used, associated with expression of type I collagen, osteoblast-associated alkaline phosphatase and osteoclastic-related tartrate-resistant acid phosphatase. Results demonstrate the potential of DPSC-loaded-dense collagen gel scaffolds to benefit of bone healing process.

Seed dormancy, seedling establishment and dynamics of the soil seed bank of Stipa bungeana (Poaceae on the Loess Plateau of northwestern China.

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Xiao Wen Hu

Full Text Available Studying seed dormancy and its consequent effect can provide important information for vegetation restoration and management. The present study investigated seed dormancy, seedling emergence and seed survival in the soil seed bank of Stipa bungeana, a grass species used in restoration of degraded land on the Loess Plateau in northwest China. Dormancy of fresh seeds was determined by incubation of seeds over a range of temperatures in both light and dark. Seed germination was evaluated after mechanical removal of palea and lemma (hulls, chemical scarification and dry storage. Fresh and one-year-stored seeds were sown in the field, and seedling emergence was monitored weekly for 8 weeks. Furthermore, seeds were buried at different soil depths, and then retrieved every 1 or 2 months to determine seed dormancy and seed viability in the laboratory. Fresh seeds (caryopses enclosed by palea and lemma had non-deep physiological dormancy. Removal of palea and lemma, chemical scarification, dry storage (afterripening, gibberellin (GA3 and potassium nitrate (KNO3 significantly improved germination. Dormancy was completely released by removal of the hulls, but seeds on which hulls were put back to their original position germinated to only 46%. Pretreatment of seeds with a 30% NaOH solution for 60 min increased germination from 25% to 82%. Speed of seedling emergence from fresh seeds was significantly lower than that of seeds stored for 1 year. However, final percentage of seedling emergence did not differ significantly for seeds sown at depths of 0 and 1 cm. Most fresh seeds of S. bungeana buried in the field in early July either had germinated or lost viability by September. All seeds buried at a depth of 5 cm had lost viability after 5 months, whereas 12% and 4% seeds of those sown on the soil surface were viable after 5 and 12 months, respectively.

Tree seed rain and seed removal, but not the seed bank, impede forest recovery in bracken (Pteridium aquilinum (L.) Kuhn)-dominated clearings in the African highlands.

Science.gov (United States)

Ssali, Fredrick; Moe, Stein R; Sheil, Douglas

2018-04-01

Considerable areas dominated by bracken Pteridium aquilinum (L.) Kuhn occur worldwide and are associated with arrested forest recovery. How forest recovery is impeded in these areas remains poorly understood, especially in the African highlands. The component processes that can lead to recruitment limitation-including low seed arrival, availability and persistence-are important determinants of plant communities and offer a potential explanation for bracken persistence. We investigated key processes that can contribute to recruitment limitation in bracken-dominated clearings in the Bwindi Impenetrable National Park, Uganda. We examined if differences in seed rain (dispersal limitation), soil seed bank, or seed removal (seed viability and persistence) can, individually or in combination, explain the differences in tree regeneration found between bracken-dominated areas and the neighboring forest. These processes were assessed along ten 50-m transects crossing the forest-bracken boundary. When compared to the neighboring forest, bracken clearings had fewer seedlings (bracken 11,557 ± 5482 vs. forest 34,515 ± 6066 seedlings/ha), lower seed rain (949 ± 582 vs. 1605 ± 335 tree seeds m -2  year -1 ), comparable but sparse soil seed bank (304 ± 236 vs. 264 ± 99 viable tree seeds/m 2 ), higher seed removal (70.1% ± 2.4% vs. 40.6% ± 2.4% over a 3-day interval), and markedly higher rodent densities (25.7 ± 5.4 vs. 5.0 ± 1.6 rodents per 100 trapping sessions). Camera traps revealed that rodents were the dominant animals visiting the seeds in our seed removal study. Synthesis : Recruitment limitation contributes to both the slow recovery of forest in bracken-dominated areas, and to the composition of the tree species that occur. Low seed arrival and low persistence of unburied seeds can both explain the reduced density of seedlings found in bracken versus neighboring forest. Seed removal, likely due to rodents, in particular appears sufficient to

α-Xylosidase plays essential roles in xyloglucan remodelling, maintenance of cell wall integrity, and seed germination in Arabidopsis thaliana.

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Shigeyama, Takuma; Watanabe, Asuka; Tokuchi, Konatsu; Toh, Shigeo; Sakurai, Naoki; Shibuya, Naoto; Kawakami, Naoto

2016-10-01

Regulation and maintenance of cell wall physical properties are crucial for plant growth and environmental response. In the germination process, hypocotyl cell expansion and endosperm weakening are prerequisites for dicot seeds to complete germination. We have identified the Arabidopsis mutant thermoinhibition-resistant germination 1 (trg1), which has reduced seed dormancy and insensitivity to unfavourable conditions for germination owing to a loss-of-function mutation of TRG1/XYL1, which encodes an α-xylosidase. Compared to those of wild type, the elongating stem of trg1 showed significantly lower viscoelasticity, and the fruit epidermal cells were longitudinally shorter and horizontally enlarged. Actively growing tissues of trg1 over-accumulated free xyloglucan oligosaccharides (XGOs), and the seed cell wall had xyloglucan with a greatly reduced molecular weight. These observations suggest that XGOs reduce xyloglucan size by serving as an acceptor in transglycosylation and eventually enhancing cell wall loosening. TRG1/XYL1 gene expression was abundant in growing wild-type organs and tissues but relatively low in cells at most actively elongating part of the tissues, suggesting that α-xylosidase contributes to maintaining the mechanical integrity of the primary cell wall in the growing and pre-growing tissues. In germinating seeds of trg1, expression of genes encoding specific abscisic acid and gibberellin metabolism enzymes was altered in accordance with the aberrant germination phenotype. Thus, cell wall integrity could affect seed germination not only directly through the physical properties of the cell wall but also indirectly through the regulation of hormone gene expression. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

The biological effect of 125I seed continuous low dose rate irradiation in CL187 cells

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Zhuang Hong-Qing

2009-01-01

Full Text Available Abstract Background To investigate the effectiveness and mechanism of 125I seed continuous low-dose-rate irradiation on colonic cell line CL187 in vitro. Methods The CL187 cell line was exposed to radiation of 60Coγ ray at high dose rate of 2 Gy/min and 125I seed at low dose rate of 2.77 cGy/h. Radiation responses to different doses and dose rates were evaluated by colony-forming assay. Under 125I seed low dose rate irradiation, a total of 12 culture dishes were randomly divided into 4 groups: Control group, and 2, 5, and 10 Gy irradiation groups. At 48 h after irradiation, apoptosis was detected by Annexin and Propidium iodide (PI staining. Cell cycle arrests were detected by PI staining. In order to investigate the influence of low dose rate irradiation on the MAPK signal transduction, the expression changes of epidermal growth factor receptor (EGFR and Raf under continuous low dose rate irradiation (CLDR and/or EGFR monoclonal antibodies were determined by indirect immunofluorescence. Results The relative biological effect (RBE for 125I seeds compared with 60Co γ ray was 1.41. Apoptosis rates of CL187 cancer cells were 13.74% ± 1.63%, 32.58% ± 3.61%, and 46.27% ± 3.82% after 2 Gy, 5 Gy, and 10 Gy irradiation, respectively; however, the control group apoptosis rate was 1.67% ± 0.19%. G2/M cell cycle arrests of CL187 cancer cells were 42.59% ± 3.21%, 59.84% ± 4.96%, and 34.61% ± 2.79% after 2 Gy, 5 Gy, and 10 Gy irradiation, respectively; however, the control group apoptosis rate was 26.44% ± 2.53%. P 2/M cell cycle arrest. After low dose rate irradiation, EGFR and Raf expression increased, but when EGFR was blocked by a monoclonal antibody, EGFR and Raf expression did not change. Conclusion 125I seeds resulted in more effective inhibition than 60Co γ ray high dose rate irradiation in CL187 cells. Apoptosis following G2/M cell cycle arrest was the main mechanism of cell-killing effects under low dose rate irradiation. CLDR could

Antileukemic Potential of Momordica charantia Seed Extracts on Human Myeloid Leukemic HL60 Cells

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Ramani Soundararajan

2012-01-01

Full Text Available Momordica charantia (bitter gourd has been used in the traditional system of medicine for the treatment of various diseases. Anticancer activity of M. charantia extracts has been demonstrated by numerous in vitro and in vivo studies. In the present study, we investigated the differentiation inducing potential of fractionated M. charantia seed extracts in human myeloid HL60 cells. We found that the HL60 cells treated with the fractionated seed extracts differentiated into granulocytic lineage as characterized by NBT staining, CD11b expression, and specific esterase activity. The differentiation inducing principle was found to be heat-stable, and organic in nature. The differentiation was accompanied by a downregulation of c-myc transcript, indicating the involvement of c-myc pathway, at least in part, in differentiation. Taken together these results indicate that fractionated extracts of M. charantia seeds possess differentiation inducing activity and therefore can be evaluated for their potential use in differentiation therapy for leukemia in combination with other inducers of differentiation.

Antileukemic Potential of Momordica charantia Seed Extracts on Human Myeloid Leukemic HL60 Cells

Science.gov (United States)

Soundararajan, Ramani; Prabha, Punit; Rai, Umesh; Dixit, Aparna

2012-01-01

Momordica charantia (bitter gourd) has been used in the traditional system of medicine for the treatment of various diseases. Anticancer activity of M. charantia extracts has been demonstrated by numerous in vitro and in vivo studies. In the present study, we investigated the differentiation inducing potential of fractionated M. charantia seed extracts in human myeloid HL60 cells. We found that the HL60 cells treated with the fractionated seed extracts differentiated into granulocytic lineage as characterized by NBT staining, CD11b expression, and specific esterase activity. The differentiation inducing principle was found to be heat-stable, and organic in nature. The differentiation was accompanied by a downregulation of c-myc transcript, indicating the involvement of c-myc pathway, at least in part, in differentiation. Taken together these results indicate that fractionated extracts of M. charantia seeds possess differentiation inducing activity and therefore can be evaluated for their potential use in differentiation therapy for leukemia in combination with other inducers of differentiation. PMID:22654956

Strategies for improving production performance of probiotic Pediococcus acidilactici viable cell by overcoming lactic acid inhibition.

Science.gov (United States)

Othman, Majdiah; Ariff, Arbakariya B; Wasoh, Helmi; Kapri, Mohd Rizal; Halim, Murni

2017-11-27

Lactic acid bacteria are industrially important microorganisms recognized for fermentative ability mostly in their probiotic benefits as well as lactic acid production for various applications. Fermentation conditions such as concentration of initial glucose in the culture, concentration of lactic acid accumulated in the culture, types of pH control strategy, types of aeration mode and different agitation speed had influenced the cultivation performance of batch fermentation of Pediococcus acidilactici. The maximum viable cell concentration obtained in constant fed-batch fermentation at a feeding rate of 0.015 L/h was 6.1 times higher with 1.6 times reduction in lactic acid accumulation compared to batch fermentation. Anion exchange resin, IRA 67 was found to have the highest selectivity towards lactic acid compared to other components studied. Fed-batch fermentation of P. acidilactici coupled with lactic acid removal system using IRA 67 resin showed 55.5 and 9.1 times of improvement in maximum viable cell concentration compared to fermentation without resin for batch and fed-batch mode respectively. The improvement of the P. acidilactici growth in the constant fed-batch fermentation indicated the use of minimal and simple process control equipment is an effective approach for reducing by-product inhibition. Further improvement in the cultivation performance of P. acidilactici in fed-bath fermentation with in situ addition of anion-exchange resin significantly helped to enhance the growth of P. acidilactici by reducing the inhibitory effect of lactic acid and thus increasing probiotic production.

Dendritic cells take up and present antigens from viable and apoptotic polymorphonuclear leukocytes.

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Carlos Alfaro

Full Text Available Dendritic cells (DC are endowed with the ability to cross-present antigens from other cell types to cognate T cells. DC are poised to meet polymorphonuclear leukocytes (PMNs as a result of being co-attracted by interleukin-8 (IL-8, for instance as produced by tumor cells or infected tissue. Human monocyte-derived and mouse bone marrow-derived DC can readily internalize viable or UV-irradiated PMNs. Such internalization was abrogated at 4°C and partly inhibited by anti-CD18 mAb. In mice, DC which had internalized PMNs containing electroporated ovalbumin (OVA protein, were able to cross-present the antigen to CD8 (OT-1 and CD4 (OT-2 TCR-transgenic T cells. Moreover, in humans, tumor cell debris is internalized by PMNs and the tumor-cell material can be subsequently taken up from the immunomagnetically re-isolated PMNs by DC. Importantly, if human neutrophils had endocytosed bacteria, they were able to trigger the maturation program of the DC. Moreover, when mouse PMNs with E. coli in their interior are co-injected in the foot pad with DC, many DC loaded with fluorescent material from the PMNs reach draining lymph nodes. Using CT26 (H-2(d mouse tumor cells, it was observed that if tumor cells are intracellularly loaded with OVA protein and UV-irradiated, they become phagocytic prey of H-2(d PMNs. If such PMNs, that cannot present antigens to OT-1 T cells, are immunomagnetically re-isolated and phagocytosed by H-2(b DC, such DC productively cross-present OVA antigen determinants to OT-1 T cells. Cross-presentation to adoptively transferred OT-1 lymphocytes at draining lymph nodes also take place when OVA-loaded PMNs (H-2(d are coinjected in the footpad of mice with autologous DC (H-2(b. In summary, our results indicate that antigens phagocytosed by short-lived PMNs can be in turn internalized and productively cross-presented by DC.

Effects of seed priming and water potential on seed germination and ...

African Journals Online (AJOL)

Administrator

2011-05-23

May 23, 2011 ... (without seed drying), primed and 12 h drying on seed germination of wheat .... completely with the lower half and the paper were rolled and placed ..... water stress and control of germination: a review. Plant Cell Environ.

Tissue Equivalents Based on Cell-Seeded Biodegradable Microfluidic Constructs

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Sarah L. Tao

2010-03-01

Full Text Available One of the principal challenges in the field of tissue engineering and regenerative medicine is the formation of functional microvascular networks capable of sustaining tissue constructs. Complex tissues and vital organs require a means to support oxygen and nutrient transport during the development of constructs both prior to and after host integration, and current approaches have not demonstrated robust solutions to this challenge. Here, we present a technology platform encompassing the design, construction, cell seeding and functional evaluation of tissue equivalents for wound healing and other clinical applications. These tissue equivalents are comprised of biodegradable microfluidic scaffolds lined with microvascular cells and designed to replicate microenvironmental cues necessary to generate and sustain cell populations to replace dermal and/or epidermal tissues lost due to trauma or disease. Initial results demonstrate that these biodegradable microfluidic devices promote cell adherence and support basic cell functions. These systems represent a promising pathway towards highly integrated three-dimensional engineered tissue constructs for a wide range of clinical applications.

Proximate content and lipid profile of seeds from Rapanea melanophloeos (The Cape Beech) tree

International Nuclear Information System (INIS)

Lotier, N.; Chivandi, E.; Lembede, B.W.; Ibrahim, K.G.; Nyakudya, T.T.

2017-01-01

Rapanea melanophloeos(Cape beech)is a fruit-bearing tree indigenous to Southern Africa which produces edible fruit. Seed samples obtained from ripe fruit of the Cape beech trees had their proximate analysis and lipid profiling done. The dry matter and ash contributed 91.29 +- 0.00 %, 1.50 +- 0.01 % of the mass of the seed. Whilst the other proximate analytes namely crude fibre, crude protein and ether extract made up 5.71 +- 0.43 %, 10.50 +- 0.49 % and 4.75 +- 0.09 percentage of the mass of the seeds respectively. The fatty acid profile of the seed oil revealed Linoleic acid (50.43 +- 0.38 %) to be the most dominant. R. melanophloeos seeds are not a viable source of nutrients. (author)

Glioma Surgical Aspirate: A Viable Source of Tumor Tissue for Experimental Research

International Nuclear Information System (INIS)

Day, Bryan W.; Stringer, Brett W.; Wilson, John; Jeffree, Rosalind L.; Jamieson, Paul R.

2013-01-01

Brain cancer research has been hampered by a paucity of viable clinical tissue of sufficient quality and quantity for experimental research. This has driven researchers to rely heavily on long term cultured cells which no longer represent the cancers from which they were derived. Resection of brain tumors, particularly at the interface between normal and tumorigenic tissue, can be carried out using an ultrasonic surgical aspirator (CUSA) that deposits liquid (blood and irrigation fluid) and resected tissue into a sterile bottle for disposal. To determine the utility of CUSA-derived glioma tissue for experimental research, we collected 48 CUSA specimen bottles from glioma patients and analyzed both the solid tissue fragments and dissociated tumor cells suspended in the liquid waste fraction. We investigated if these fractions would be useful for analyzing tumor heterogeneity, using IHC and multi-parameter flow cytometry; we also assessed culture generation and orthotopic xenograft potential. Both cell sources proved to be an abundant, highly viable source of live tumor cells for cytometric analysis, animal studies and in-vitro studies. Our findings demonstrate that CUSA tissue represents an abundant viable source to conduct experimental research and to carry out diagnostic analyses by flow cytometry or other molecular diagnostic procedures

In vitro evaluation of cell-seeded chitosan films for peripheral nerve tissue engineering

OpenAIRE

Wrobel, Sandra; Serra, Sofia Cristina; Samy, S. M.; Sousa, Nuno; Heimann, Claudia; Barwig, Christina; Grothe, Claudia; Salgado, A. J.; Talini, Kirsten Haastert

2014-01-01

Natural biomaterials have attracted an increasing interest in the field of tissue-engineered nerve grafts, representing a possible alternative to autologous nerve transplantation. With the prospect of developing a novel entubulation strategy for transected nerves with cell-seeded chitosan films, we examined the biocompatibility of such films in vitro. Different types of rat Schwann cells (SCs)-immortalized, neonatal, and adult-as well as rat bone-marrow-derived mesenchymal stromal cells (BMSC...

Microfluidic monitoring of programmed cell death in living plant seed tissue

DEFF Research Database (Denmark)

Mark, Christina; Heiskanen, Arto; Zor, Kinga

, et al., (2006), BioEssays, 28, p. 1091). Microfluidic cell culture enables in vitro experiments to approach in vivo conditions. Combining microfluidics with the Lab-On-a-Chip concept allows implementing a wide range of assays for real-time monitoring of effects in a biological system of factors...... such as concentration of selected compounds, external pH, oxygen consumption, redox state and cell viability. The aleurone layer of the barley seed is a 2-3 single cell type thick tissue that can be dissected from the embryo and starchy endosperm. During incubation in vitro this mechanically very robust maintains...

Comparison of Cuminaldehyde Contents from Cell Suspension Cultures and Seeds of [Bunium persicum (Boiss. B. Fedtsch.

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Sara KHOSRAVINIA

2012-11-01

Full Text Available The cell suspension culture and seed samples of Bunium persicum were extracted by supercritical fluid, hydrodistillation and solvent methods and analyzed by Gas Chromatography. In this study to compare the different methods of extractions, cuminaldehyde was targeted as one of the Black zira essential oil constitute. For callus induction the germinated seeds were cultured as explants on Murashige and Skoog medium supplemented with 2 mg/l 2,4-dichlorophenoxy acetic acid and 0.5 mg/l kinetin (treatment A as well as 2 mg/l ?-naphthalene acetic acid and 0.5 mg/l 6-benzyl aminopurine (treatment B and followed by cells suspension cultures establishment for the first time. The results of cell culture showed that cells from treatment B have a growth rate higher than A. All extracts were dissolved in 1 ml hexane and analyzed by Gas Chromatography. According to the Gas Chromatography analysis, cuminaldehyde was not detected in the supercritical fluid samples, while it was present in hydrodistillation and solvent extract. Cuminaldehyde percentage in cell and seed solvent extracts was 4.65% and 18.61% respectively. Gas Chromatography results also showed that no cuminaldehyde is present in media extracts, means no cuminaldehyde has been secreted into the medium.

Extremely long-distance seed dispersal by an overfished Amazonian frugivore.

Science.gov (United States)

Anderson, Jill T; Nuttle, Tim; Saldaña Rojas, Joe S; Pendergast, Thomas H; Flecker, Alexander S

2011-11-22

Throughout Amazonia, overfishing has decimated populations of fruit-eating fishes, especially the large-bodied characid, Colossoma macropomum. During lengthy annual floods, frugivorous fishes enter vast Amazonian floodplains, consume massive quantities of fallen fruits and egest viable seeds. Many tree and liana species are clearly specialized for icthyochory, and seed dispersal by fish may be crucial for the maintenance of Amazonian wetland forests. Unlike frugivorous mammals and birds, little is known about seed dispersal effectiveness of fishes. Extensive mobility of frugivorous fish could result in extremely effective, multi-directional, long-distance seed dispersal. Over three annual flood seasons, we tracked fine-scale movement patterns and habitat use of wild Colossoma, and seed retention in the digestive tracts of captive individuals. Our mechanistic model predicts that Colossoma disperses seeds extremely long distances to favourable habitats. Modelled mean dispersal distances of 337-552 m and maximum of 5495 m are among the longest ever reported. At least 5 per cent of seeds are predicted to disperse 1700-2110 m, farther than dispersal by almost all other frugivores reported in the literature. Additionally, seed dispersal distances increased with fish size, but overfishing has biased Colossoma populations to smaller individuals. Thus, overexploitation probably disrupts an ancient coevolutionary relationship between Colossoma and Amazonian plants.

Ethical issues in the use of the terminator seed technology | Yusuf ...

African Journals Online (AJOL)

Biodiversity and food security especially in developing countries are under threat by a newly patented technique for preventing plants from producing viable seeds. The method dubbed “the suicide seeds” by critics was developed in a joint venture between the United State Department of Agriculture (USDA) and Mississippi ...

COBRA-LIKE2, a member of the glycosylphosphatidylinositol-anchored COBRA-LIKE family, plays a role in cellulose deposition in arabidopsis seed coat mucilage secretory cells.

Science.gov (United States)

Ben-Tov, Daniela; Abraham, Yael; Stav, Shira; Thompson, Kevin; Loraine, Ann; Elbaum, Rivka; de Souza, Amancio; Pauly, Markus; Kieber, Joseph J; Harpaz-Saad, Smadar

2015-03-01

Differentiation of the maternally derived seed coat epidermal cells into mucilage secretory cells is a common adaptation in angiosperms. Recent studies identified cellulose as an important component of seed mucilage in various species. Cellulose is deposited as a set of rays that radiate from the seed upon mucilage extrusion, serving to anchor the pectic component of seed mucilage to the seed surface. Using transcriptome data encompassing the course of seed development, we identified COBRA-LIKE2 (COBL2), a member of the glycosylphosphatidylinositol-anchored COBRA-LIKE gene family in Arabidopsis (Arabidopsis thaliana), as coexpressed with other genes involved in cellulose deposition in mucilage secretory cells. Disruption of the COBL2 gene results in substantial reduction in the rays of cellulose present in seed mucilage, along with an increased solubility of the pectic component of the mucilage. Light birefringence demonstrates a substantial decrease in crystalline cellulose deposition into the cellulosic rays of the cobl2 mutants. Moreover, crystalline cellulose deposition into the radial cell walls and the columella appears substantially compromised, as demonstrated by scanning electron microscopy and in situ quantification of light birefringence. Overall, the cobl2 mutants display about 40% reduction in whole-seed crystalline cellulose content compared with the wild type. These data establish that COBL2 plays a role in the deposition of crystalline cellulose into various secondary cell wall structures during seed coat epidermal cell differentiation. © 2015 American Society of Plant Biologists. All Rights Reserved.

Osmoconditioning prevents the onset of microtubular cytoskeleton and activation of cell cycle and is detrimental for germination of Jatropha curcas L. seeds.

Science.gov (United States)

de Brito, C D; Loureiro, M B; Ribeiro, P R; Vasconcelos, P C T; Fernandez, L G; de Castro, R D

2016-11-01

Jatropha curcas is an oilseed crop renowned for its tolerance to a diverse range of environmental stresses. In Brazil, this species is grown in semiarid regions where crop establishment requires a better understanding of the mechanisms underlying appropriate seed, seedling and plant behaviour under water restriction conditions. In this context, the objective of this study was to investigate the physiological and cytological profiles of J. curcas seeds in response to imbibition in water (control) and in polyethylene glycol solution (osmoticum). Seed germinability and reactivation of cell cycle events were assessed by means of different germination parameters and immunohistochemical detection of tubulin and microtubules, i.e. tubulin accumulation and microtubular cytoskeleton configurations in water imbibed seeds (control) and in seeds imbibed in the osmoticum. Immunohistochemical analysis revealed increasing accumulation of tubulin and appearance of microtubular cytoskeleton in seed embryo radicles imbibed in water from 48 h onwards. Mitotic microtubules were only visible in seeds imbibed in water, after radicle protrusion, as an indication of cell cycle reactivation and cell proliferation, with subsequent root development. Imbibition in osmoticum prevented accumulation of microtubules, i.e. activation of cell cycle, therefore germination could not be resumed. Osmoconditioned seeds were able to survive re-drying and could resume germination after re-imbibition in water, however, with lower germination performance, possibly due to acquisition of secondary dormancy. This study provides important insights into understanding of the physiological aspects of J. curcas seed germination in response to water restriction conditions. © 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.

The biomechanics of seed germination.

Science.gov (United States)

Steinbrecher, Tina; Leubner-Metzger, Gerhard

2017-02-01

From a biomechanical perspective, the completion of seed (and fruit) germination depends on the balance of two opposing forces: the growth potential of the embryonic axis (radicle-hypocotyl growth zone) and the restraint of the seed-covering layers (endosperm, testa, and pericarp). The diverse seed tissues are composite materials which differ in their dynamic properties based on their distinct cell wall composition and water uptake capacities. The biomechanics of embryo cell growth during seed germination depend on irreversible cell wall loosening followed by water uptake due to the decreasing turgor, and this leads to embryo elongation and eventually radicle emergence. Endosperm weakening as a prerequisite for radicle emergence is a widespread phenomenon among angiosperms. Research into the biochemistry and biomechanics of endosperm weakening has demonstrated that the reduction in puncture force of a seed's micropylar endosperm is environmentally and hormonally regulated and involves tissue-specific expression of cell wall remodelling proteins such as expansins, diverse hydrolases, and the production of directly acting apoplastic reactive oxygen. The endosperm-weakening biomechanics and its underlying cell wall biochemistry differ between the micropylar (ME) and chalazal (CE) endosperm domains. In the ME, they involve cell wall loosening, cell separation, and programmed cell death to provide decreased and localized ME tissue resistance, autolysis, and finally the formation of an ME hole required for radicle emergence. Future work will further unravel the molecular mechanisms, environmental regulation, and evolution of the diverse biomechanical cell wall changes underpinning the control of germination by endosperm weakening. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Mutagenic effects of gamma rays on the frequency and spectrum of chlorophyll and viable macro mutations in chilli (Capsicum annuum L.)

International Nuclear Information System (INIS)

Rangaiah, S.; Manjunath, A.; Puttaramanaik

2004-01-01

In the present study, uniform dry and healthy seeds of two cultivars of chilli namely Ceylon and Byadagi were exposed to 60 Co gamma rays at 5, 10, 15, 20, 25, 30, 35 and 40 krad doses at BARC, Mumbai. Thus there were total 18 treatments, eight different doses of gamma rays and one control in each cultivar. The M2 populations were compared with the respective control populations for chlorophyll and viable mutations. The treated populations were compared with the respective control populations for chlorophyll mutations (M2 and M3) and viable mutations (M2)' The results of the study are presented

Galápagos land iguana (Conolophus subcristatus) as a seed disperser.

Science.gov (United States)

Traveset, Anna; Nogales, Manuel; Vargas, Pablo; Rumeu, Beatriz; Olesen, Jens M; Jaramillo, Patricia; Heleno, Ruben

2016-05-01

The role of the most common land iguana (Conolophus subcristatus) in the Galápagos Islands as an effective seed disperser is explored in this study. A total of 5705 seeds of 32 plant species were identified from 160 scats, 4545 of which (80%) appeared visually undamaged. Germination trials of 849 seeds from 29 species revealed that at least 10 species remained viable after passing through the iguana's gut, although only a small proportion of those seeds (4%) germinated. In any case, we argue that C. subcristatus exerts an important role on the 7 Galapagos islands where it occurs because of its abundance and capacity to ingest and disperse seeds at long distances. Our results strongly suggest that the Galápagos C. subcristatus plays an important role as a seed disperser of not only of native species but also some introduced plants in the Galápagos Islands. © 2016 International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.

Designing primers and evaluation of the efficiency of propidium monoazide – Quantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius

Directory of Open Access Journals (Sweden)

Chieh-Hsien Lai

2017-07-01

Full Text Available The purpose of this study is to evaluate the efficiency of using propidium monoazide (PMA real-time quantitative polymerase chain reaction (qPCR to count the viable cells of Lactobacillus gasseri and Lactobacillus salivarius in probiotic products. Based on the internal transcription spacer and 23S rRNA genes, two primer sets specific for these two Lactobacillus species were designed. For a probiotic product, the total deMan Rogosa Sharpe plate count was 8.65±0.69 log CFU/g, while for qPCR, the cell counts of L. gasseri and L. salivarius were 8.39±0.14 log CFU/g and 8.57±0.24 log CFU/g, respectively. Under the same conditions, for its heat-killed product, qPCR counts for L. gasseri and L. salivarius were 6.70±0.16 log cells/g and 7.67±0.20 log cells/g, while PMA-qPCR counts were 5.33±0.18 log cells/g and 5.05±0.23 log cells/g, respectively. For cell dilutions with a viable cell count of 8.5 log CFU/mL for L. gasseri and L. salivarius, after heat killing, the PMA-qPCR count for both Lactobacillus species was near 5.5 log cells/mL. When the PMA-qPCR counts of these cell dilutions were compared before and after heat killing, although some DNA might be lost during the heat killing, significant qPCR signals from dead cells, i.e., about 4–5 log cells/mL, could not be reduced by PMA treatment. Increasing PMA concentrations from 100 μM to 200 μM or light exposure time from 5 minutes to 15 minutes had no or, if any, only minor effect on the reduction of qPCR signals from their dead cells. Thus, to differentiate viable lactic acid bacterial cells from dead cells using the PMA-qPCR method, the efficiency of PMA to reduce the qPCR signals from dead cells should be notable.

Ricinosomes provide an early indicator of suspensor and endosperm cells destined to die during late seed development in quinoa (Chenopodium quinoa).

Science.gov (United States)

López-Fernández, M P; Maldonado, S

2013-11-01

In mature quinoa (Chenopodium quinoa) seeds, the lasting endosperm forms a micropylar cone covering the radicle. The suspensor cells lie within the centre of the cone. During the final stage of seed development, the cells of the lasting endosperm accumulate protein and lipids while the rest are crushed and disintegrated. Both the suspensor and endosperm die progressively from the innermost layers surrounding the embryo and extending towards the nucellar tissue. Ricinosomes are endoplasmic reticulum-derived organelles that accumulate both the pro-form and the mature form of cysteine endopeptidase (Cys-EP), first identified in castor bean (Ricinus communis) endosperm during germination. This study sought to identify associations between the presence of ricinosomes and programmed cell death (PCD) hallmarks in suspensor and endosperm cells predestined to die during quinoa seed development. A structural study using light microscopy and transmission electron microscopy was performed. To detect the presence of Cys-EP, both western blot and in situ immunolocalization assays were carried out using anti-R. communis Cys-EP antibody. A TUNEL assay was used to determine DNA fragmentation. Except for the one or two cell layers that constitute the lasting endosperm in the mature seed, ricinosomes were found in suspensor and endosperm cells. These cells were also the site of morphological abnormalities, including misshapen and fragmented nuclei, vesiculation of the cytosol, vacuole collapse and cell wall disorganization. It is proposed that, in suspensor and endosperm cells, the early detection of Cys-EP in ricinosomes predicts the occurrence of PCD during late seed development.

Oxidative Stress Induces Senescence in Cultured RPE Cells.

Science.gov (United States)

Aryan, Nona; Betts-Obregon, Brandi S; Perry, George; Tsin, Andrew T

2016-01-01

The aim of this research is to determine whether oxidative stress induces cellular senescence in human retinal pigment epithelial cells. Cultured ARPE19 cells were subjected to different concentrations of hydrogen peroxide to induce oxidative stress. Cells were seeded into 24-well plates with hydrogen peroxide added to cell medium and incubated at 37°C + 5% CO2 for a 90-minute period [at 0, 300, 400 and 800 micromolar (MCM) hydrogen peroxide]. The number of viable ARPE19 cells were recorded using the Trypan Blue Dye Exclusion Method and cell senescence was measured by positive staining for senescence-associated beta-galactosidase (SA-beta-Gal) protein. Without hydrogen peroxide treatment, the number of viable ARPE19 cells increased significantly from 50,000 cells/well to 197,000 within 72 hours. Treatment with hydrogen peroxide reduced this level of cell proliferation significantly (to 52,167 cells at 400 MCM; to 49,263 cells at 800 MCM). Meanwhile, cells with a high level of positive senescence-indicator SA-Beta-Gal-positive staining was induced by hydrogen peroxide treatment (from a baseline level of 12% to 80% at 400 MCM and at 800 MCM). Our data suggests that oxidative stress from hydrogen peroxide treatment inhibited ARPE19 cell proliferation and induced cellular senescence.

Pepper seed germination assessed by combined X-radiography and computer-aided imaging analysis

International Nuclear Information System (INIS)

Dell'Aquila, A.

2007-01-01

A lot of pepper seeds having 87% germination were subjected to X-ray inspection using a non lethal dose of radiation. Seeds with less than 2.7% (on the basis of total seed area) of free space area, i.e. the spaces between embryo and endosperm, were classified as highly viable seeds (97-100% germination) with the lowest level of abnormal seedlings. Seeds X-ray classified as good were subjected to a computerised image analysis to study seed imbibition and radicle elongation. The patterns of seed area increase, chosen as the most accurate indicator of seed swelling, resembled the triphasic curve of water uptake. The first phase was completed at 9 h followed by a second phase that varied widely in time until completion of germination between 52 and 96 h. The proportion of seeds with radicle protrusion between 52-56 h and 64-72 h assessed with the image analysis was significantly higher than that recorded using a conventional germination test. In addition, the rate of increase of seed area during the third phase of imbibition, mostly due to protrusion of the radicle tip and its growth, was highly correlated with the corresponding radicle elongation rate

Characterization of PDF1 and its interaction with DELAY OF GERMINATION1 (DOG1) in the control of seed dormancy in Arabidopsis thaliana

OpenAIRE

Miatton, Emma

2012-01-01

Seed dormancy is defined as the incapacity of a viable seed to germinate under favourable conditions. It is established during seed maturation and reaches high levels in mature dry seeds. Dormancy is a complex adaptive trait that assures germination at proper time of the year at the onset of the favourable growing season. This trait is regulated by hormonal and environmental cues such as temperature and light. In Arabidopsis thaliana dormancy can be released by imbibing seeds at cold temperat...

Designing primers and evaluation of the efficiency of propidium monoazide - Quantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius.

Science.gov (United States)

Lai, Chieh-Hsien; Wu, Sih-Rong; Pang, Jen-Chieh; Ramireddy, Latha; Chiang, Yu-Cheng; Lin, Chien-Ku; Tsen, Hau-Yang

2017-07-01

The purpose of this study is to evaluate the efficiency of using propidium monoazide (PMA) real-time quantitative polymerase chain reaction (qPCR) to count the viable cells of Lactobacillus gasseri and Lactobacillus salivarius in probiotic products. Based on the internal transcription spacer and 23S rRNA genes, two primer sets specific for these two Lactobacillus species were designed. For a probiotic product, the total deMan Rogosa Sharpe plate count was 8.65±0.69 log CFU/g, while for qPCR, the cell counts of L. gasseri and L. salivarius were 8.39±0.14 log CFU/g and 8.57±0.24 log CFU/g, respectively. Under the same conditions, for its heat-killed product, qPCR counts for L. gasseri and L. salivarius were 6.70±0.16 log cells/g and 7.67±0.20 log cells/g, while PMA-qPCR counts were 5.33±0.18 log cells/g and 5.05±0.23 log cells/g, respectively. For cell dilutions with a viable cell count of 8.5 log CFU/mL for L. gasseri and L. salivarius, after heat killing, the PMA-qPCR count for both Lactobacillus species was near 5.5 log cells/mL. When the PMA-qPCR counts of these cell dilutions were compared before and after heat killing, although some DNA might be lost during the heat killing, significant qPCR signals from dead cells, i.e., about 4-5 log cells/mL, could not be reduced by PMA treatment. Increasing PMA concentrations from 100 μM to 200 μM or light exposure time from 5 minutes to 15 minutes had no or, if any, only minor effect on the reduction of qPCR signals from their dead cells. Thus, to differentiate viable lactic acid bacterial cells from dead cells using the PMA-qPCR method, the efficiency of PMA to reduce the qPCR signals from dead cells should be notable. Copyright © 2016. Published by Elsevier B.V.

Entry of Vibrio harveyi and Vibrio fischeri into the viable but nonculturable state

Digital Repository Service at National Institute of Oceanography (India)

Ramaiah, Neelam; Ravel, J.; Straube, W.L.; Hill, R.T.; Colwell, R.R.

, direct viable counts, actively respiring cell counts, nucleoid-containing cell counts, and total counts were determined. V. harveyi incubated at 22 degrees C nutrient-limited artificial seawater (ASW) became nonculturable after approximately 62 and 45 d...

Seed coat development in Velloziaceae: primary homology assessment and insights on seed coat evolution.

Science.gov (United States)

Sousa-Baena, Mariane S; de Menezes, Nanuza L

2014-09-01

• Seed coat characteristics have historically been used to infer taxonomic relationships and are a potential source of characters for phylogenetic reconstruction. In particular, seed coat morphoanatomy has never been studied in detail in Velloziaceae. One character based on seed surface microsculpture has been used in phylogenies, but was excluded from recent studies owing to problems in primary homology. This work aimed to clarify the origin and general composition of seed coat cell layers in Velloziaceae and to propose hypotheses of primary homology among seed characters.• Seed coat development of 24 Velloziaceae species, comprising nine genera, and one species of Pandanaceae (outgroup) was studied using standard anatomical methods. Developmental data were interpreted in the light of a recently published phylogeny.• Eight types of seed coat were identified. Whereas the most common type has four distinct cell layers (two-layered tegmen and testa), we encountered much more variation in seed coat composition than previously reported, the analysis of which revealed some potential synapomorphies. For instance, an exotesta with spiral thickenings may be a synapomorphy of Barbacenia.• Our results showed that the character states previously used in phylogenies are not based on homologous layers and that the same state was misattributed to species exhibiting quite different seed coats. This study is a first step toward a better understanding of seed coat structure evolution in Velloziaceae. © 2014 Botanical Society of America, Inc.

Viability of small seeds found in feces of the Central American tapir on Barro Colorado Island, Panama.

Science.gov (United States)

Capece, Paula I; Aliaga-Rossel, Enzo; Jansen, Patrick A

2013-03-01

Tapirs are known as effective dispersers of large-seeded tree species, but their role in dispersing small-seeded plant species has yet to be established. Tapir feces have been reported to contain large numbers of small seeds, but whether these are viable has rarely been evaluated. We determined the abundance and viability of small seeds in feces of Central American tapir (Tapirus bairdii) on Barro Colorado Island, Panama. A total of 72 fecal samples were collected opportunistically from 4 tapir latrine sites. Seeds were manually extracted from feces and classified by size. Seed viability was estimated by opening each seed and examining for the presence of at least 1 intact firm white endosperm. In total, we obtained 8166 seeds of at least 16 plant species. Small-seeded species dominated, with 96% of all seeds found measuring tapirs potentially serve as effective dispersers of a wide range of small-seeded plant species. © 2012 Wiley Publishing Asia Pty Ltd, ISZS and IOZ/CAS.

A polysaccharide fraction of adlay seed (Coixlachryma-jobi L.) induces apoptosis in human non-small cell lung cancer A549 cells

Energy Technology Data Exchange (ETDEWEB)

Lu, Xiangyi; Liu, Wei; Wu, Junhua; Li, Mengxian [Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457 (China); Wang, Juncheng; Wu, Jihui [School of Life Science, University of Science and Technology of China, Hefei 230022 (China); Luo, Cheng, E-mail: Luo58@yahoo.com [Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457 (China)

2013-01-11

Highlights: Black-Right-Pointing-Pointer A polysaccharide from adlay seed, its molecular mass, optical rotation and sugars was determined. Black-Right-Pointing-Pointer We demonstrated that a polysaccharide from adlay can induce apoptosis in cancer cells. Black-Right-Pointing-Pointer The polysaccharide inhibited the metabolism and proliferation of NSCLC A549 cells. Black-Right-Pointing-Pointer The polysaccharide may trigger apoptosis via the mitochondria-dependent pathway. -- Abstract: Different seed extracts from Coix lachryma-jobi (adlay seed) have been used for the treatment of various cancers in China, and clinical data support the use of these extracts for cancer therapy; however, their underlying molecular mechanisms have not been well defined. A polysaccharide fraction, designated as CP-1, was extracted from the C.lachryma-jobi L. var. using the ethanol subsiding method. CP-1 induced apoptosis in A549 cells in a dose-dependent manner, as determined by MTT assay. Apoptotic bodies were observed in the cells by scanning electronic microscopy. Apoptosis and DNA accumulation during S-phase of the cell cycle were determined by annexin V-FITC and PI staining, respectively, and measured by flow cytometry. CP-1 also extended the comet tail length on single cell gel electrophoresis, and disrupted the mitochondrial membrane potential. Further analysis by western blotting showed that the expression of caspase-3 and caspase-9 proteins was increased. Taken together, our results demonstrate that CP-1 is capable of inhibiting A549 cell proliferation and inducing apoptosis via a mechanism primarily involving the activation of the intrinsic mitochondrial pathway. The assay data suggest that in addition to its nutritional properties, CP-1 is a very promising candidate polysaccharide for the development of anti-cancer medicines.

A polysaccharide fraction of adlay seed (Coixlachryma-jobi L.) induces apoptosis in human non-small cell lung cancer A549 cells

International Nuclear Information System (INIS)

Lu, Xiangyi; Liu, Wei; Wu, Junhua; Li, Mengxian; Wang, Juncheng; Wu, Jihui; Luo, Cheng

2013-01-01

Highlights: ► A polysaccharide from adlay seed, its molecular mass, optical rotation and sugars was determined. ► We demonstrated that a polysaccharide from adlay can induce apoptosis in cancer cells. ► The polysaccharide inhibited the metabolism and proliferation of NSCLC A549 cells. ► The polysaccharide may trigger apoptosis via the mitochondria-dependent pathway. -- Abstract: Different seed extracts from Coix lachryma-jobi (adlay seed) have been used for the treatment of various cancers in China, and clinical data support the use of these extracts for cancer therapy; however, their underlying molecular mechanisms have not been well defined. A polysaccharide fraction, designated as CP-1, was extracted from the C.lachryma-jobi L. var. using the ethanol subsiding method. CP-1 induced apoptosis in A549 cells in a dose-dependent manner, as determined by MTT assay. Apoptotic bodies were observed in the cells by scanning electronic microscopy. Apoptosis and DNA accumulation during S-phase of the cell cycle were determined by annexin V-FITC and PI staining, respectively, and measured by flow cytometry. CP-1 also extended the comet tail length on single cell gel electrophoresis, and disrupted the mitochondrial membrane potential. Further analysis by western blotting showed that the expression of caspase-3 and caspase-9 proteins was increased. Taken together, our results demonstrate that CP-1 is capable of inhibiting A549 cell proliferation and inducing apoptosis via a mechanism primarily involving the activation of the intrinsic mitochondrial pathway. The assay data suggest that in addition to its nutritional properties, CP-1 is a very promising candidate polysaccharide for the development of anti-cancer medicines.

A Classification Method for Seed Viability Assessment with Infrared Thermography

Directory of Open Access Journals (Sweden)

Sen Men

2017-04-01

Full Text Available This paper presents a viability assessment method for Pisum sativum L. seeds based on the infrared thermography technique. In this work, different artificial treatments were conducted to prepare seeds samples with different viability. Thermal images and visible images were recorded every five minutes during the standard five day germination test. After the test, the root length of each sample was measured, which can be used as the viability index of that seed. Each individual seed area in the visible images was segmented with an edge detection method, and the average temperature of the corresponding area in the infrared images was calculated as the representative temperature for this seed at that time. The temperature curve of each seed during germination was plotted. Thirteen characteristic parameters extracted from the temperature curve were analyzed to show the difference of the temperature fluctuations between the seeds samples with different viability. With above parameters, support vector machine (SVM was used to classify the seed samples into three categories: viable, aged and dead according to the root length, the classification accuracy rate was 95%. On this basis, with the temperature data of only the first three hours during the germination, another SVM model was proposed to classify the seed samples, and the accuracy rate was about 91.67%. From these experimental results, it can be seen that infrared thermography can be applied for the prediction of seed viability, based on the SVM algorithm.

Anatomy and cell wall polysaccharides of almond (Prunus dulcis D. A. Webb) seeds.

Science.gov (United States)

Dourado, Fernando; Barros, António; Mota, Manuel; Coimbra, Manuel A; Gama, Francisco M

2004-03-10

The anatomy of Prunus dulcis was analyzed by applying several differential staining techniques and light microscopy. Prunus dulcis seed has a thin and structurally complex seed coat, with lignified cellulosic tissue. The embryo has two voluminous cotyledons. Cotyledon cells have a high number of protein and lipid bodies, some of which have phytin. The provascular tissue, located in the cotyledons, is oriented in small bundles perpendicular to the transverse embryonic axis. Prunus dulcis cell wall material is very rich in arabinose (45 mol %). Glucose (23%), uronic acids (12%), and xylose (12%) are also major sugar components. The polymers obtained from the imidazole and Na(2)CO(3) extracts contain mainly pectic substances rich in arabinose, but the sugar content of these extracts was very low. The majority of the pectic substances (also rich in arabinose) was recovered with the KOH extracts. These extracts, with high sugar content, yielded also xyloglucans and acidic xylans. The 4 M KOH + H(3)BO(3) extracts yielded polysaccharides rich in uronic acids and xylose and very rich in arabinose, accounting for 27% of the cell wall material.

Seed dormancy and persistent sediment seed banks of ephemeral freshwater rock pools in the Australian monsoon tropics.

Science.gov (United States)

Cross, Adam T; Turner, Shane R; Renton, Michael; Baskin, Jerry M; Dixon, Kingsley W; Merritt, David J

2015-04-01

Rock pools are small, geologically stable freshwater ecosystems that are both hydrologically and biologically isolated. They harbour high levels of plant endemism and experience environmental unpredictability driven by the presence of water over variable temporal scales. This study examined the hypothesis that the sediment seed bank in monsoon tropical freshwater rock pools would persist through one or more periods of desiccation, with seed dormancy regulating germination timing in response to rock pool inundation and drying events. Seeds were collected from seven dominant rock pool species, and germination biology and seed dormancy were assessed under laboratory conditions in response to light, temperature and germination stimulators (gibberellic acid, karrikinolide and ethylene). Field surveys of seedling emergence from freshwater rock pools in the Kimberley region of Western Australia were undertaken, and sediment samples were collected from 41 vegetated rock pools. Seedling emergence and seed bank persistence in response to multiple wetting and drying cycles were determined. The sediment seed bank of individual rock pools was large (13 824 ± 307 to 218 320 ± 42 412 seeds m(-2) for the five species investigated) and spatially variable. Seedling density for these same species in the field ranged from 13 696 to 87 232 seedlings m(-2). Seeds of rock pool taxa were physiologically dormant, with germination promoted by after-ripening and exposure to ethylene or karrikinolide. Patterns of seedling emergence varied between species and were finely tuned to seasonal temperature and moisture conditions, with the proportions of emergent seedlings differing between species through multiple inundation events. A viable seed bank persisted after ten consecutive laboratory inundation events, and seeds retained viability in dry sediments for at least 3 years. The persistent seed bank in freshwater rock pools is likely to provide resilience to plant

Factors affecting post-control reinvasion by seed of an invasive species, Phragmites australis, in the central Platte River, Nebraska.

Science.gov (United States)

Galatowitsch, Susan M.; Larson, Diane L.; Larson, Jennifer L.

2016-01-01

Invasive plants, such as Phragmites australis, can profoundly affect channel environments of large rivers by stabilizing sediments and altering water flows. Invasive plant removal is considered necessary where restoration of dynamic channels is needed to provide critical habitat for species of conservation concern. However, these programs are widely reported to be inefficient. Post-control reinvasion is frequent, suggesting increased attention is needed to prevent seed regeneration. To develop more effective responses to this invader in the Central Platte River (Nebraska, USA), we investigated several aspects of Phragmites seed ecology potentially linked to post-control reinvasion, in comparison to other common species: extent of viable seed production, importance of water transport, and regeneration responses to hydrology. We observed that although Phragmites seed does not mature until very late in the ice-free season, populations produce significant amounts of viable seed (>50 % of filled seed). Most seed transported via water in the Platte River are invasive perennial species, although Phragmites abundances are much lower than species such as Lythrum salicaria, Cyperus esculentus and Phalaris arundinacea. Seed regeneration of Phragmites varies greatly depending on hydrology, especially timing of water level changes. Flood events coinciding with the beginning of seedling emergence reduced establishment by as much as 59 % compared to flood events that occurred a few weeks later. Results of these investigations suggest that prevention of seed set (i.e., by removal of flowering culms) should be a priority in vegetation stands not being treated annually. After seeds are in the seedbank, preventing reinvasion using prescribed flooding has a low chance of success given that Phragmites can regenerate in a wide variety of hydrologic microsites.

Co-Seeding Human Endothelial Cells with Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells on Calcium Phosphate Scaffold Enhances Osteogenesis and Vascularization in Rats.

Science.gov (United States)

Liu, Xian; Chen, Wenchuan; Zhang, Chi; Thein-Han, Wahwah; Hu, Kevin; Reynolds, Mark A; Bao, Chongyun; Wang, Ping; Zhao, Liang; Xu, Hockin H K

2017-06-01

A major challenge in repairing large bone defects with tissue-engineered constructs is the poor vascularization in the defect. The lack of vascular networks leads to insufficient oxygen and nutrients supply, which compromises the survival of seeded cells. To achieve favorable regenerative effects, prevascularization of tissue-engineered constructs by co-culturing of endothelial cells and bone cells is a promising strategy. The aim of this study was to investigate the effects of human-induced pluripotent stem cell-derived mesenchymal stem cells (hiPSC-MSCs) co-cultured with human umbilical vein endothelial cells (HUVECs) for prevascularization of calcium phosphate cement (CPC) scaffold on bone regeneration in vivo for the first time. HUVECs co-cultured with hiPSC-MSCs formed microcapillary-like structures in vitro. HUVECs promoted mineralization of hiPSC-MSCs on CPC scaffolds. Four groups were tested in a cranial bone defect model in nude rats: (1) CPC scaffold alone (CPC control); (2) HUVEC-seeded CPC (CPC-HUVEC); (3) hiPSC-MSC-seeded CPC (CPC-hiPSC-MSC); and (4) HUVECs co-cultured with hiPSC-MSCs on CPC scaffolds (co-culture group). After 12 weeks, the co-culture group achieved the greatest new bone area percentage of 46.38% ± 3.8% among all groups (p < 0.05), which was more than four folds of the 10.61% ± 1.43% of CPC control. In conclusion, HUVECs co-cultured with hiPSC-MSCs substantially promoted bone regeneration. The novel construct of HUVECs co-cultured with hiPSC-MSCs delivered via CPC scaffolds is promising to enhance bone and vascular regeneration in orthopedic applications.

Increasing seed size and quality by manipulating BIG SEEDS1 in legume species.

Science.gov (United States)

Ge, Liangfa; Yu, Jianbin; Wang, Hongliang; Luth, Diane; Bai, Guihua; Wang, Kan; Chen, Rujin

2016-11-01

Plant organs, such as seeds, are primary sources of food for both humans and animals. Seed size is one of the major agronomic traits that have been selected in crop plants during their domestication. Legume seeds are a major source of dietary proteins and oils. Here, we report a conserved role for the BIG SEEDS1 (BS1) gene in the control of seed size and weight in the model legume Medicago truncatula and the grain legume soybean (Glycine max). BS1 encodes a plant-specific transcription regulator and plays a key role in the control of the size of plant organs, including seeds, seed pods, and leaves, through a regulatory module that targets primary cell proliferation. Importantly, down-regulation of BS1 orthologs in soybean by an artificial microRNA significantly increased soybean seed size, weight, and amino acid content. Our results provide a strategy for the increase in yield and seed quality in legumes.

Reduced germination success of temperate grassland seeds sown in dung: consequences for post-dispersal seed fate.

Science.gov (United States)

Milotić, T; Hoffmann, M

2016-11-01

Endozoochory is one of the main drivers shaping temperate grassland communities by maintaining plant populations of its constituents and enabling plants to colonize new habitats. Successful endozoochorous dispersal implies that seeds not only get consumed and survive the digestive tract but are also able to develop into viable seedlings in a dung environment. We experimentally assessed the germination probability and timing of 15 annual and perennial temperate European grassland species in cattle and horse dung and in different climatic conditions (greenhouse and outdoor conditions). Interspecific variation in germinability and germination timing are found, while life strategy had only an effect on germination timing. We found adverse effects of both cattle and horse dung on the germination characteristics of all tested grassland species, but the effects of cattle dung were more pronounced. In comparison with the control treatment, fewer seeds emerged in dung and more time was needed to germinate. Also, germination metrics clearly differed between the artificial greenhouse and outdoor conditions, with generally a lower germinability in outdoor conditions. According to our results, a large cost seems to be associated with endozoochorous dispersal in this stage of the life cycle, as seed dispersal effectiveness strongly depends on the quality of the deposition site with a lowered survival and germination probability when seeds are deposited in dung. © 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.

Construction of a system using a deep learning algorithm to count cell numbers in nanoliter wells for viable single-cell experiments.

Science.gov (United States)

Kamatani, Takashi; Fukunaga, Koichi; Miyata, Kaede; Shirasaki, Yoshitaka; Tanaka, Junji; Baba, Rie; Matsusaka, Masako; Kamatani, Naoyuki; Moro, Kazuyo; Betsuyaku, Tomoko; Uemura, Sotaro

2017-12-04

For single-cell experiments, it is important to accurately count the number of viable cells in a nanoliter well. We used a deep learning-based convolutional neural network (CNN) on a large amount of digital data obtained as microscopic images. The training set consisted of 103 019 samples, each representing a microscopic grayscale image. After extensive training, the CNN was able to classify the samples into four categories, i.e., 0, 1, 2, and more than 2 cells per well, with an accuracy of 98.3% when compared to determination by two trained technicians. By analyzing the samples for which judgments were discordant, we found that the judgment by technicians was relatively correct although cell counting was often difficult by the images of discordant samples. Based on the results, the system was further enhanced by introducing a new algorithm in which the highest outputs from CNN were used, increasing the accuracy to higher than 99%. Our system was able to classify the data even from wells with a different shape. No other tested machine learning algorithm showed a performance higher than that of our system. The presented CNN system is expected to be useful for various single-cell experiments, and for high-throughput and high-content screening.

TRANSPLANTATION OF CRYOPRESERVED FETAL LIVER CELLS SEEDED INTO MACROPOROUS ALGINATE-GELATIN SCAFFOLDS IN RATS WITH LIVER FAILURE

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D. V. Grizay

2015-01-01

Full Text Available Aim. To study the therapeutic potential of cryopreserved fetal liver cells seeded into macroporous alginategelatin scaffolds after implantation to omentum of rats with hepatic failure.Materials and methods.Hepatic failure was simulated by administration of 2-acetyl aminofl uorene followed partial hepatectomy. Macroporous alginate-gelatin scaffolds, seeded with allogenic cryopreserved fetal liver cells (FLCs were implanted into rat omentum. To prevent from colonization of host cells scaffolds were coated with alginate gel shell. Serum transaminase activity, levels of albumin and bilirubin as markers of hepatic function were determined during 4 weeks after failure model formation and scaffold implantation. Morphology of liver and scaffolds after implantation were examined histologically. Results. Macroporous alginate-gelatin scaffolds after implantation to healthy rats were colonized by host cells. Additional formation of alginate gel shell around scaffolds prevented the colonization. Implantation of macroporous scaffolds seeded with cryopreserved rat FLCs and additionally coated with alginate gel shell into omentum of rats with hepatic failure resulted in signifi cant improvement of hepatospecifi c parameters of the blood serum and positive changes of liver morphology. The presence of cells with their extracellular matrix within the scaffolds was confi rmed after 4 weeks post implantation.Conclusion. The data above indicate that macroporous alginate-gelatin scaffolds coated with alginate gel shell are promising cell carriers for the development of bioengineered liver equivalents.

Seed research for improved technologies

OpenAIRE

Bino,R.J.; Jalink,H.; Oluoch,M.O.; Groot,S.P.C.

1998-01-01

The production of high-quality seed is the basis for a durable a profitable agriculture. After production, seed is processed, conditioned, stored, shipped and germinated. For quality assurance, seed quality has to be controlled at all steps of the production chain. Seed functioning is accompanied by programmed transitions from cell proliferation to quiescence upon maturation and from quiescence to reinitiation of cellular metabolism upon imbibition. Despite the obvious importance of these con...

Bioactivity of microencapsulated soursop seeds extract on Plutella xylostella

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Ismael Barros Gomes

2016-05-01

Full Text Available ABSTRACT: The aim of this study was to evaluate the bioactivity of microencapsulated extract from the soursop seeds, Annona muricata L. ( Annonaceae , on diamondback moth, Plutella xylostela L. (Lepidoptera: Plutellidae . Microencapsulation was performed in a Mini Spray Dryer model B-290 using 50mL of ethanolic and hexanic extracts plus 150mL of ethanol and 150mL of ultrapure water, mixed with aerosil (first polymer or arabic gum (second polymer. It was possible to microencapsulate the ethanolic extract of soursop seeds only by using the polymer arabic gum at 20%. The microencapsulated extract caused significant acute toxicity (LC50=258mg L-1 and chronic effects, especially reduction of larval viability and increased larval stage. We concluded that the microencapsulation of the ethanolic extract of soursop seeds can be a viable alternative for controlling diamondback moth with possible gains for the environment.

Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns.

Science.gov (United States)

Seth, Gargi; Hamilton, Robert W; Stapp, Thomas R; Zheng, Lisa; Meier, Angela; Petty, Krista; Leung, Stephenie; Chary, Srikanth

2013-05-01

Agility to schedule and execute cell culture manufacturing campaigns quickly in a multi-product facility will play a key role in meeting the growing demand for therapeutic proteins. In an effort to shorten campaign timelines, maximize plant flexibility and resource utilization, we investigated the initiation of cell culture manufacturing campaigns using CHO cells cryopreserved in large volume bags in place of the seed train process flows that are conventionally used in cell culture manufacturing. This approach, termed FASTEC (Frozen Accelerated Seed Train for Execution of a Campaign), involves cultivating cells to high density in a perfusion bioreactor, and cryopreserving cells in multiple disposable bags. Each run for a manufacturing campaign would then come from a thaw of one or more of these cryopreserved bags. This article reviews the development and optimization of individual steps of the FASTEC bioprocess scheme: scaling up cells to greater than 70 × 10(6) cells/mL and freezing in bags with an optimized controlled rate freezing protocol and using a customized rack configuration. Flow cytometry analysis was also employed to understand the recovery of CHO cells following cryopreservation. Extensive development data were gathered to ensure that the quantity and quality of the drug manufactured using the FASTEC bioprocess scheme was acceptable compared to the conventional seed train process flow. The result of offering comparable manufacturing options offers flexibility to the cell culture manufacturing network. Copyright © 2012 Wiley Periodicals, Inc.

CHANGES IN NUTRIENT AND ANTINUTRITIONAL CONTENTS OF SESAME SEEDS DURING FERMENTATION

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Aderonke I. Olagunju

2013-06-01

Full Text Available Sesame seeds were fermented using the traditional method for four days and samples taken for analysis each day till the last day of fermentation to monitor the compositional changes in the seeds as fermentation progressed. The viable count obtained ranged from 8.0×103 after 24 h to 2.93×106 cfu/g on the 4th day. The crude protein and fat content increased as fermentation progressed reaching 27.84% and 51.58% respectively. Fermentation yielded positive effect on the phytic acid, phytin phosphorus and oxalate content of the flour samples when compared with the control. Phytic acid content ranged from 31.59 mg/g for raw seed to 18.13 mg/g for fermented seed flour. Sesame seed are high in minerals such as calcium, magnesium, potassium, sodium and slight increases in values were obtained at the end of processing. Sesame seeds are rich in both essential and non-essential amino acids with leucine, methionine, phenylalanine, threonine and valine values higher than the recommended daily allowance. Processing significantly increased the amino acid values. Sesame flour demonstrated ability to scavenge free radicals. Fermentation of sesame seeds resulted in reduction in the antinutrients in the seed and the seed can serve as soup condiment and seasoning with improved nutritional composition with respect to protein and amino acid.

Radioactive ceramic seeds with Ho-166 and Sm-153 with perspective of use in brachytherapy

International Nuclear Information System (INIS)

Valente, Eduardo Sarmento

2010-01-01

This work aimed to characterize physically, chemically and radioactively ceramics seeds with holmium and samarium embedded in ceramic matrix produced by sol-gel route. Biological response of HeLa cells was investigated macroscopically in vitro under the exposure to β radiation of Ho-166 and Sm-153. The dispersal of radioactive material in simulated body fluid (SBF) was made and the dose rate and total dose deposited in the volume delimited by the range of β particles in the water was calculated. The methodology used in physical, chemical and nuclear characterization was based on the following techniques: atomic emission spectrometry, neutron activation analysis, gamma spectrometry, X-ray fluorescence spectrometry and X-rays diffraction. In addition to the solubility in SBF, an in vivo pilot study assessed the dispersion of radioactive seeds implanted in rabbit muscle tissue. Four experiments were prepared to investigate the biological response in vitro, in which radioactive seeds were placed directly in contact with the cells in confluent culture flasks. The first two were done with seed's activity of 3.7 MBq (0.1 mCi) for the Sm-153 and 11.1 MBq (0.3 mCi) for Ho-166. There were no concerns with spatial positioning for seeds in those early experiments. The subsequent experiments were performed with activity ranging from 37 MBq (1 mCi) and 111 MBq (3 mCi) and with defined position of the seeds in the flask. The results showed the presence of halos with maximum diameter of 8 mm around the seeds where there was a drastic reduction in the number of viable cells as a result of the effects of β radiation. The calculation of dose deposited in water as well as the dose rate calculation of during handling of the seeds for an implant followed the methodology established by MIRD. These calculations found dose rates ranging from 7 to 19 Gy/h and integrated doses between 93 and 730 Gy, due to β radiation, for 6 half-lives of radioisotopes Sm-153 and Ho-166. The dose

The differentiation of embryonic stem cells seeded on electrospun nanofibers into neural lineages.

Science.gov (United States)

Xie, Jingwei; Willerth, Stephanie M; Li, Xiaoran; Macewan, Matthew R; Rader, Allison; Sakiyama-Elbert, Shelly E; Xia, Younan

2009-01-01

Due to advances in stem cell biology, embryonic stem (ES) cells can be induced to differentiate into a particular mature cell lineage when cultured as embryoid bodies. Although transplantation of ES cells-derived neural progenitor cells has been demonstrated with some success for either spinal cord injury repair in small animal model, control of ES cell differentiation into complex, viable, higher ordered tissues is still challenging. Mouse ES cells have been induced to become neural progenitors by adding retinoic acid to embryoid body cultures for 4 days. In this study, we examine the use of electrospun biodegradable polymers as scaffolds not only for enhancing the differentiation of mouse ES cells into neural lineages but also for promoting and guiding the neurite outgrowth. A combination of electrospun fiber scaffolds and ES cells-derived neural progenitor cells could lead to the development of a better strategy for nerve injury repair.

Dye-sensitized solar cells with natural dyes extracted from achiote seeds

Energy Technology Data Exchange (ETDEWEB)

Gomez-Ortiz, N.M.; Vazquez-Maldonado, I.A.; Azamar-Barrios, J.A.; Oskam, G. [Departamento de Fisica Aplicada, CINVESTAV-IPN, Merida, Yuc. 97310 (Mexico); Perez-Espadas, A.R.; Mena-Rejon, G.J. [Laboratorio de Quimica Organica de Investigacion, Facultad de Quimica, Universidad Autonoma de Yucatan, Merida, Yuc. 97150 (Mexico)

2010-01-15

We have explored the application of natural dyes extracted from the seeds of the achiote shrub (Bixa orellana L.) in dye-sensitized solar cells (DSCs). The main pigments are bixin and norbixin, which were obtained by separation and purification from the dark-red extract (annatto). The dyes were characterized using {sup 1}H-NMR, FTIR spectroscopy, and UV-Vis spectrophotometry. Solar cells were prepared using TiO{sub 2} and ZnO nanostructured, mesoporous films and the annatto, bixin, and norbixin as sensitizers. The best results were obtained with bixin-sensitized TiO{sub 2} solar cells with efficiencies of up to 0.53%, illustrating the importance of purification of dyes from natural extracts. (author)

Banco de sementes de arroz vermelho em sistemas de semeadura de arroz irrigado Red rice seed bank under seeding systems of flooded rice

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Luis Antonio de Avila

2000-10-01

Full Text Available O experimento foi conduzido com o objetivo de estimar a variação do banco de sementes viáveis de arroz vermelho (Oryza sativa L., em três sistemas de semeadura de arroz (O. sativa L. irrigado (semeadura convencional, direta e com sementes pré-germinadas, após três anos de cultivo na mesma área, em São Borja, RS, em solo classificado como Planossolo, com infestação média de 150 plantas de arroz vermelho por m² antes da instalação do experimento. A coleta do solo para estimativa do banco de sementes de arroz vermelho foi realizada com um trado cilíndrico de metal de 10cm de diâmetro. Foram realizadas amostragens de solo, com 12 amostras por parcela em duas profundidades (0-1cm e 1-10cm, para retirar os grãos de arroz vermelho, submetê-los ao teste de tetrazólio e determinar sua viabilidade. O experimento foi um bifatorial em blocos ao acaso, com três repetições, com os sistemas de semeadura de arroz como fator principal e as profundidades de coleta como fator secundário. Os dados demonstraram que o número de sementes viáveis de arroz vermelho no solo foi maior na área onde o arroz foi cultivado no sistema de semeadura convencional (1994 sementes/m², com valores intermediários para o sistema de semeadura direta (597 sementes/m² e menores valores para o sistema de semeadura com sementes pré-germinadas (93 sementes/m². Assim, demonstra-se que a semeadura de arroz no sistema de sementes pré-germinadas favorece o controle do arroz vermelho, mantendo a quantidade de sementes em densidades menores do que nos sistemas de semeadura direta e convencional.An experiment was conducted aiming to estimate the variability of viable seeds of red rice (Oryza sativa L. in soil under three rice (O. sativa L. seeding systems: conventional, no-tillage and water seeded. It was located in São Borja, Rio Grande do Sul state, Brazil, in an albaqualf soil with an average of 150 red rice plants per square meter before establishing the

Bystander Effects Induced by Continuous Low-Dose-Rate 125I Seeds Potentiate the Killing Action of Irradiation on Human Lung Cancer Cells In Vitro

International Nuclear Information System (INIS)

Chen, H.H.; Jia, R.F.; Yu, L.; Zhao, M.J.; Shao, C.L.; Cheng, W.Y.

2008-01-01

Purpose: To investigate bystander effects of low-dose-rate (LDR) 125 I seed irradiation on human lung cancer cells in vitro. Methods and Materials: A549 and NCI-H446 cell lines of differing radiosensitivity were directly exposed to LDR 125 I seeds irradiation for 2 or 4 Gy and then cocultured with nonirradiated cells for 24 hours. Induction of micronucleus (MN), γH2AX foci, and apoptosis were assayed. Results: After 2 and 4 Gy irradiation, micronucleus formation rate (MFR) and apoptotic rate of A549 and NCI-H446 cells were increased, and the MFR and apoptotic rate of NCI-H446 cells was 2.1-2.8 times higher than that of A549 cells. After coculturing nonirradiated bystander cells with 125 I seed irradiated cells for 24 hours, MFR and the mean number of γH2AX foci/cells of bystander A549 and NCI-H446 cells were similar and significantly higher than those of control (p 125 I seeds could induce bystander effects, which potentiate the killing action on tumor cells and compensate for the influence of nonuniform distribution of radiation dosage on therapeutic outcomes

The Conceptual Mechanism for Viable Organizational Learning Based on Complex System Theory and the Viable System Model

Science.gov (United States)

Sung, Dia; You, Yeongmahn; Song, Ji Hoon

2008-01-01

The purpose of this research is to explore the possibility of viable learning organizations based on identifying viable organizational learning mechanisms. Two theoretical foundations, complex system theory and viable system theory, have been integrated to provide the rationale for building the sustainable organizational learning mechanism. The…

Fate of viable but non-culturable Listeria monocytogenes in pig manure microcosms

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Jeremy eDesneux

2016-03-01

Full Text Available The fate of two strains of L. monocytogenes and their ability to become viable but non-culturable (VBNC was investigated in microcosms containing piggery effluents (two raw manures and two biologically treated manures stored for two months at 8°C and 20°C. Levels of L. monocytogenes were estimated using the culture method, qPCR, and propidium monoazide treatment combined with qPCR (qPCRPMA. The chemical composition and the microbial community structure of the manures were also analysed. The strains showed similar decline rates and persisted up to 63 days. At day zero, the percentage of VBNC cells among viable cells was higher in raw manures (81.5-94.8% than in treated manures (67.8-79.2%. The changes in their proportion over time depended on the temperature and on the type of effluent: the biggest increase was observed in treated manures at 20°C and the smallest increase in raw manures at 8°C. The chemical parameters had no influence on the behaviour of the strains, but decrease of the persistence of viable cells was associated with an increase in the microbial richness of the manures. This study demonstrated that storing manure altered the culturability of L. monocytogenes, which rapidly entered the VBNC state, and underlines the importance of including VBNC cells when estimating the persistence of the pathogens in farm effluents.

Establishment of post-harvest early-developmental categories for viability maintenance of Araucaria angustifolia seeds

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Cristhyane Garcia Araldi

2015-12-01

Full Text Available Araucaria angustifolia seeds are recalcitrant, and their metabolism remains high during storage. This research aimed to describe the initiation of germination in A. angustifolia seeds during storage in order to standardize the assessment of physiological quality and to promote seed conservation. Seeds were collected from two populations and stored for 270 days in the natural laboratory environment and cold chamber. Seeds were classified according to four early developmental stages: I - mature seeds; II - seeds with elongation along the embryonic axis; III - beginning of root protrusion; IV - advanced germination stage, with seedling shoots. After categorization, physical and physiological quality was assessed. In freshly collected seeds, only category I was observed. At 270 days, approximately 40% of seeds were in category III in laboratory conditions, while the maintenance in a cold chamber delayed germinative metabolism. Viability tests showed that seeds in categories III and IV were more susceptible to damage caused by storage. In conclusion, the percentage of viable A. angustifolia seeds depends on the development stage after collection. Seeds that have reached early developmental category III should be prioritized for propagation, while those remaining in categories I and II should be longer stored with periodic assessment for reduction in physiological quality.

X-radiation-induced transformation in a C3H mouse embryo-derived cell line

International Nuclear Information System (INIS)

Terzaghi, M.; Little, J.B.

1976-01-01

Reproducible x-ray-induced oncogenic transformation has been demonstrated in an established cell line of mouse embryo fibroblasts. Cells derived from transformed foci formed malignant tumors when injected into syngeneic hosts. An exponential increase in the number of transformants per viable cell occurred with doses of up to 400 rads of x-radiation. The transformation frequency in exponentially growing cultures remained constant at 2.3 x 10 -3 following doses of 400 to 1500 rads. There was little change in survival following x-ray doses up to 300 rads. Doses greater than 300 rads were associated with an exponential decline in survival; the D 0 for the survival curve was 175 rads. Transformation frequency varied with changes in the number of viable cells seeded per dish. There was about a 10-fold decline in the transformation frequency when the number of cells was increased from 400 to 1000 viable cells/100-mm Petri dish. Below this density range there was little change in transformation frequency. The presence of lethally preirradiated cells was not associated with an enhancement of transformation in irradiated cells or with the induction of transformation in unirradiated cell cultures. Amphotericin B (Fungizone) inhibited the appearance of transformants when added to the culture medium within 2 to 3 weeks after initiation of the experiment

Ligament Tissue Engineering Using a Novel Porous Polycaprolactone Fumarate Scaffold and Adipose Tissue-Derived Mesenchymal Stem Cells Grown in Platelet Lysate.

Science.gov (United States)

Wagner, Eric R; Bravo, Dalibel; Dadsetan, Mahrokh; Riester, Scott M; Chase, Steven; Westendorf, Jennifer J; Dietz, Allan B; van Wijnen, Andre J; Yaszemski, Michael J; Kakar, Sanjeev

2015-11-01

Surgical reconstruction of intra-articular ligament injuries is hampered by the poor regenerative potential of the tissue. We hypothesized that a novel composite polymer "neoligament" seeded with progenitor cells and growth factors would be effective in regenerating native ligamentous tissue. We synthesized a fumarate-derivative of polycaprolactone fumarate (PCLF) to create macro-porous scaffolds to allow cell-cell communication and nutrient flow. Clinical grade human adipose tissue-derived human mesenchymal stem cells (AMSCs) were cultured in 5% human platelet lysate (PL) and seeded on scaffolds using a dynamic bioreactor. Cell growth, viability, and differentiation were examined using metabolic assays and immunostaining for ligament-related markers (e.g., glycosaminoglycans [GAGs], alkaline phosphatase [ALP], collagens, and tenascin-C). AMSCs seeded on three-dimensional (3D) PCLF scaffolds remain viable for at least 2 weeks with proliferating cells filling the pores. AMSC proliferation rates increased in PL compared to fetal bovine serum (FBS) (p ligament and tenogenic growth factor fibroblast growth factor 2 (FGF-2), especially when cultured in the presence of PL (p engineering and ligament regeneration.

Effect of GA3 treatment on seed development and seed-related gene expression in grape.

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Chenxia Cheng

Full Text Available The phytohormone gibberellic acid (GA3 is widely used in the table grape industry to induce seedlessness in seeded varieties. However, there is a paucity of information concerning the mechanisms by which GAs induce seedlessness in grapes.In an effort to systematically analyze the cause of this GA3-induced seed abortion, we conducted an in depth characterization of two seeded grape cultivars ('Kyoho' and 'Red Globe', along with a seedless cultivar ('Thompson Seedless', following treatment with GA3. In a similar fashion to the seedless control, which exhibited GA3-induced abortion of the seeds 9 days after full bloom (DAF, both 'Kyoho' and 'Red Globe' seeded varieties exhibited complete abortion of the seeds 15 DAF when treated with GA3. Morphological analyses indicated that while fertilization appeared to occur normally following GA3 treatment, as well as in the untreated seedless control cultivar, seed growth eventually ceased. In addition, we found that GA3 application had an effect on redox homeostasis, which could potentially cause cell damage and subsequent seed abortion. Furthermore, we carried out an analysis of antioxidant enzyme activities, as well as transcript levels from various genes believed to be involved in seed development, and found several differences between GA3-treated and untreated controls.Therefore, it seems that the mechanisms driving GA3-induced seedlessness are similar in both seeded and seedless cultivars, and that the observed abortion of seeds may result at least in part from a GA3-induced increase in cell damage caused by reactive oxygen species, a decrease in antioxidant enzymatic activities, and an alteration of the expression of genes related to seed development.

Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum

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Ching Giap Tan

2014-09-01

Full Text Available The present study was based on the reverse transcription polymerase chain reaction (RT-PCR of the 16S ribosomal nucleic acid (rRNA of Mycoplasma for detection of viable Mycoplasma gallisepticum. To determine the stability of M. gallisepticum 16S rRNA in vitro, three inactivation methods were used and the suspensions were stored at different temperatures. The 16S rRNA of M. gallisepticum was detected up to approximately 20–25 h at 37 °C, 22–25 h at 16 °C, and 23–27 h at 4 °C. The test, therefore, could detect viable or recently dead M. gallisepticum (< 20 h. The RT-PCR method was applied during an in vivo study of drug efficacy under experimental conditions, where commercial broiler-breeder eggs were inoculated with M. gallisepticum into the yolk. Hatched chicks that had been inoculated in ovo were treated with Macrolide 1. The method was then applied in a flock of day 0 chicks with naturally acquired vertical transmission of M. gallisepticum, treated with Macrolide 2. Swabs of the respiratory tract were obtained for PCR and RT-PCR evaluations to determine the viability of M. gallisepticum. This study proved that the combination of both PCR and RT-PCR enables detection and differentiation of viable from non-viable M. gallisepticum.

Monitoring viable cells of the biological control agent Lactobacillus plantarum PM411 in aerial plant surfaces by means of a strain-specific viability quantitative PCR.

Science.gov (United States)

Daranas, Núria; Bonaterra, Anna; Francés, Jesús; Cabrefiga, Jordi; Montesinos, Emilio; Badosa, Esther

2018-03-09

A viability qPCR (v-qPCR) assay was developed for the unambiguous detection and quantification of Lactobacillus plantarum PM411 viable cells in aerial plant surfaces. A 972 bp region of a PM411 predicted prophage with mosaic architecture enabled the identification of a PM411 strain-specific molecular marker. Three primer sets, with different amplicon lengths (92, 188, and 317 bp), and one TaqMan probe were designed. All the qPCR assays showed good linearity over a 4-log range and good efficiencies, but differed in sensitivity. The nucleic acid-binding dye PEMAX was used for selectively detecting and enumerating viable bacteria by v-qPCR. The primer set amplifying a 188 bp DNA fragment was selected as the most suitable for v-qPCR. The performance of the method was assessed on apple blossoms, pear, strawberry and kiwifruit leaves in potted plants under controlled environmental conditions, and pear and apple blossoms under field conditions, by comparing v-qPCR population estimation to those obtained by qPCR and specific plate counting on MRS-rifampicin. The population estimation did not differ significantly between methods when conditions were conducive to bacterial survival. However, under stressful conditions, differences between methods were observed due to cell death or viable but non-culturable state induction. While qPCR overestimated the population level, plate counting underestimated this value in comparison to v-qPCR. PM411 attained stable population levels of viable cells on flower environment under high relative humidity. However, the unfavourable conditions onto the leaf surface and the relatively dryness in the field caused an important decrease of viable population. IMPORTANCE The v-qPCR method in combination with plate counting and qPCR is a powerful tool for studies of colonization and survival in field conditions, to improve formulations and delivery strategies of PM411, or to optimize the dose and timing of spray schedules. It is expected that PEMAX

Evaluation of PCR and DNA hybridization protocols for detection of viable enterotoxigenic Clostridium perfringens in irradiated beef

International Nuclear Information System (INIS)

Baez, L.A.; Juneja, V.K.; Thayer, D.W.; Sackitey, S.

1997-01-01

The sensitivity of DNA hybridization and polymerase chain reaction (PCR), was evaluated in irradiated cooked and raw beef samples. A membrane-based colony hybridization assay and a PCR protocol, both with specificity for the enterotoxin A gene of Clostridium perfringens, were compared with viable plate counts. The results of the colony hybridization procedure were in agreement with viable plate counts for detection and enumeration of enterotoxigenic C. perfringens. The PCR procedure combined a 4 h enrichment followed by a nucleic acid extraction step and assessed the amplification of 183 and 750 base pair enterotoxin gene targets. Detection of C. perfringens by PCR did not show a reliable correlation with viable plate counts or the colony hybridization assay. C. perfringens killed by irradiation were not detected by the plate count or colony hybridization methods; however, killed cells were detected with the PCR technique. By relying on the growth of viable cells for detection and/or enumeration, the colony hybridization and plate count methods provided a direct correlation with the presence of viable bacteria

Portulaca oleracea Seed Oil Exerts Cytotoxic Effects on Human Liver Cancer (HepG2) and Human Lung Cancer (A-549) Cell Lines.

Science.gov (United States)

Al-Sheddi, Ebtesam Saad; Farshori, Nida Nayyar; Al-Oqail, Mai Mohammad; Musarrat, Javed; Al-Khedhairy, Abdulaziz Ali; Siddiqui, Maqsood Ahmed

2015-01-01

Portulaca oleracea (Family: Portulacaceae), is well known for its anti-inflammatory, antioxidative, anti- bacterial, and anti-tumor activities. However, cytotoxic effects of seed oil of Portulaca oleracea against human liver cancer (HepG2) and human lung cancer (A-549) cell lines have not been studied previously. Therefore, the present study was designed to investigate the cytotoxic effects of Portulaca oleracea seed oil on HepG2 and A-549 cell lines. Both cell lines were exposed to various concentrations of Portulaca oleracea seed oil for 24h. After the exposure, percentage cell viability was studied by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT), neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed a concentration-dependent significant reduction in the percentage cell viability and an alteration in the cellular morphology of HepG2 and A-549 cells. The percentage cell viability was recorded as 73%, 63%, and 54% by MTT assay and 76%, 61%, and 50% by NRU assay at 250, 500, and 1000 μg/ml, respectively in HepG2 cells. Percentage cell viability was recorded as 82%, 72%, and 64% by MTT assay and 83%, 68%, and 56% by NRU assay at 250, 500, and 1000 μg/ml, respectively in A-549 cells. The 100 μg/ml and lower concentrations were found to be non cytotoxic to A-549 cells, whereas decrease of 14% and 12% were recorded by MTT and NRU assay, respectively in HepG2 cells. Both HepG2 and A-549 cell lines exposed to 250, 500, and 1000 μg/ ml of Portulaca oleracea seed oil lost their normal morphology, cell adhesion capacity, become rounded, and appeared smaller in size. The data from this study showed that exposure to seed oil of Portulaca oleracea resulted in significant cytotoxicity and inhibition of growth of the human liver cancer (HepG2) and human lung cancer (A-549) cell lines.

Estimation of cancerolytic properties of thionine from plants seeds by inclusion of C14-thymidine in tumoral cells

International Nuclear Information System (INIS)

Pshenichnov, E.A.; Sultanova, E.M.; Kuznetsova, N.N.; Khashimova, Z.S.; Veshkurova, O.N.; Sadikov, A.A.; Salikhov, Sh.I.

2004-01-01

Full text: It has been earlier shown that cysteine rich peptides - thionine from seeds of various plants possess expressed fungitoxic activity. It is connected to influence of thionine on cellular membranes of fungi. It was possible to assume that the substances showing cytotoxic activity will be active in relation to tumoral cells. We isolated peptide fractions from seeds bamia (Hibiscus esculentus), kenaf (Hibiscus cannabinus), abutilon (Abutilon theophrasti), euphorbia (Euphorbia virgata), palma Christi (Ricinus communis) and horse sorrel (Rumex confertus) and studied their antineoplastic and fungitoxic activity. Antiproliferative action of peptides to melanoma cells of mice was estimated in cytotoxic test by inclusion of C 14 -thymidine to DNA. This researches have shown that peptides from seeds of horse sorrel and palma Christi did not change a level of synthesis of DNA while peptides from euphorbia and bamia considerably reduced inclusion of labeled nucleotide to DNA and suppressed growth of tumoral cells on 14 and 39 % accordingly. Parallel tests of these peptides on fungitoxic activity in relation to virulent strains of Verticillium dahliae have shown suppression of conidial growth on 17 and 26 % accordingly. Thus, peptides from seeds of bamia and euphorbia possess the expressed property to suppress growth of tumoral cells and can be used at creation a new cancerolytic preparations for treatment of human cancer. Work is executed under the financial support of fundamental grants F - 4.19 and F-4.1.44

Biocatalytically active silCoat-composites entrapping viable Escherichia coli.

Science.gov (United States)

Findeisen, A; Thum, O; Ansorge-Schumacher, M B

2014-02-01

Application of whole cells in industrial processes requires high catalytic activity, manageability, and viability under technical conditions, which can in principle be accomplished by appropriate immobilization. Here, we report the identification of carrier material allowing exceptionally efficient adsorptive binding of Escherichia coli whole cells hosting catalytically active carbonyl reductase from Candida parapsilosis (CPCR2). With the immobilizates, composite formation with both hydrophobic and hydrophilized silicone was achieved, yielding advanced silCoat-material and HYsilCoat-material, respectively. HYsilCoat-whole cells were viable preparations with a cell loading up to 400 mg(E. coli) · g(-1)(carrier) and considerably lower leaching than native immobilizates. SilCoat-whole cells performed particularly well in neat substrate exhibiting distinctly increased catalytic activity.

Beneficial Effect of Jojoba Seed Extracts on Hyperglycemia-Induced Oxidative Stress in RINm5f Beta Cells

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Sahla Belhadj

2018-03-01

Full Text Available Hyperglycemia occurs during diabetes and insulin resistance. It causes oxidative stress by increasing reactive oxygen species (ROS levels, leading to cellular damage. Polyphenols play a central role in defense against oxidative stress. In our study, we investigated the antioxidant properties of simmondsin, a pure molecule present in jojoba seeds, and of the aqueous extract of jojoba seeds on fructose-induced oxidative stress in RINm5f beta cells. The exposure of RINm5f beta cells to fructose triggered the loss of cell viability (−48%, p < 0.001 and disruption of insulin secretion (p < 0.001 associated with of reactive oxygen species (ROS production and a modulation of pro-oxidant and antioxidant signaling pathway. Cell pre-treatments with extracts considerably increased cell viability (+86% p < 0.001 for simmondsin and +74% (p < 0.001 for aqueous extract and insulin secretion. The extracts also markedly decreased ROS (−69% (p < 0.001 for simmondsin and −59% (p < 0.001 for aqueous extract and caspase-3 activation and improved antioxidant defense, inhibiting p22phox and increasing nuclear factor (erythroid-derived 2-like 2 (Nrf2 levels (+70%, p < 0.001 for aqueous extract. Simmondsin had no impact on Nrf2 levels. The richness and diversity of molecules present in jojoba seed extract makes jojoba a powerful agent to prevent the destruction of RINm5f beta cells induced by hyperglycemia.

Thousand-seed weight in the theory and practice

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Mirić Mladen

2007-01-01

Full Text Available Out of approximately 100 seed traits, 20 are of the marketing importance, and 10-15, including the 1000-seed weight (TSW have been studied. There are many articles partially dealing with TSW in our literature related to the seed industry, but none of them is all-inclusive. Therefore, the aim of the present study was to determine: (1 a role, (2 factors of formation and (3 obtaining desirable TWS (DTWS. Germination, yield and trade depend on TWS, while TWS is interrelated with a dozen of other parameters, selection of sowing plates, determination of the amounts of seeds necessary for sowing (sowing rate and the depth of sowing, and can be an indicator of some other seed or crop traits. TSW depends on inheritance and conditions and is interrelated with sowing, drying and processing. Individual factors of TSW are numerous and classified into two groups each with two subgroups: a agro ecological (abiotic and biotic and b agro technological (cropping practices and processing. The species and variety, crop density and uniformity, irrigation regime and fertilization have the greatest impact on TSW, while the effects of soil tillage and weather conditions, except during pollination and maturing, are of less importance. Seed fractioning is crucial, but it is less performed in our country than world-wide. The desirable TSW (DTWS is a term designating higher-quality, more homogenous and greater yield of seed units in the whole process of the seed production. It is necessary to analyze DTWS for all varieties in order to determine DTWS that will provide the highest yield of viable and uniform seeds per hectare. Labor rationalization and productivity require obtaining DTWS as high as possible in the filed and maintaining it in processing. .

COBRA-LIKE2, a Member of the Glycosylphosphatidylinositol-Anchored COBRA-LIKE Family, Plays a Role in Cellulose Deposition in Arabidopsis Seed Coat Mucilage Secretory Cells1,2[OPEN

Science.gov (United States)

Ben-Tov, Daniela; Abraham, Yael; Stav, Shira; Thompson, Kevin; Loraine, Ann; Elbaum, Rivka; de Souza, Amancio; Pauly, Markus; Kieber, Joseph J.; Harpaz-Saad, Smadar

2015-01-01

Differentiation of the maternally derived seed coat epidermal cells into mucilage secretory cells is a common adaptation in angiosperms. Recent studies identified cellulose as an important component of seed mucilage in various species. Cellulose is deposited as a set of rays that radiate from the seed upon mucilage extrusion, serving to anchor the pectic component of seed mucilage to the seed surface. Using transcriptome data encompassing the course of seed development, we identified COBRA-LIKE2 (COBL2), a member of the glycosylphosphatidylinositol-anchored COBRA-LIKE gene family in Arabidopsis (Arabidopsis thaliana), as coexpressed with other genes involved in cellulose deposition in mucilage secretory cells. Disruption of the COBL2 gene results in substantial reduction in the rays of cellulose present in seed mucilage, along with an increased solubility of the pectic component of the mucilage. Light birefringence demonstrates a substantial decrease in crystalline cellulose deposition into the cellulosic rays of the cobl2 mutants. Moreover, crystalline cellulose deposition into the radial cell walls and the columella appears substantially compromised, as demonstrated by scanning electron microscopy and in situ quantification of light birefringence. Overall, the cobl2 mutants display about 40% reduction in whole-seed crystalline cellulose content compared with the wild type. These data establish that COBL2 plays a role in the deposition of crystalline cellulose into various secondary cell wall structures during seed coat epidermal cell differentiation. PMID:25583925

The durative use of suspension cells and callus for volatile oil by comparative with seeds and fruits in Capparis spinosa L.

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Yongtai Yin

Full Text Available Capparis spinosa is one of the most important eremophytes among the medicinal plants, and continued destruction of these plants poses a major threat to species survival. The development of methods to extract compounds, especially those of medicinal value, without harvesting the whole plant is an issue of considerable socioeconomic importance. On the basis of an established system for culture of suspension cells and callus in vitro, Gas Chromatograph-Mass Spectrometer (GC-MS was used for the volatile oil composition analyzing in seed, fruit, suspension cells and callus. Fatty acids were the major component, and the highest content of alkanes was detected in seed, with <1.0% in suspension cells and callus. Esters, olefins and heterocyclic compounds were significantly higher in fruit than in the other materials. The content of acid esters in the suspension cells and callus was significantly higher than in seed and fruit. This indicated that the suspension cells and callus could be helpful for increasing the value of volatile oil and replacing seeds and fruit partially as a source of some compounds of the volatile oil and may also produce some new medical compounds. The above results give valuable information for sustainable use of C. spinosa and provide a foundation for use of the C. spinosa suspension cells and callus as an ongoing medical resource.

Spatiotemporal Dynamics of Total Viable Vibrio spp. in a NW Mediterranean Coastal Area.

Science.gov (United States)

Girard, Léa; Peuchet, Sébastien; Servais, Pierre; Henry, Annabelle; Charni-Ben-Tabassi, Nadine; Baudart, Julia

2017-09-27

A cellular approach combining Direct Viable Counting and Fluorescent In Situ Hybridization using a one-step multiple-probe technique and Solid Phase Cytometry (DVC-FISH-SPC) was developed to monitor total viable vibrios and cover the detection of a large diversity of vibrios. FISH combined three probes in the same assay and targeted sequences located at different positions on the 16S rRNA of Vibrio and Aliivibrio members. We performed a 10-month in situ study to investigate the weekly dynamics of viable vibrios relative to culturable counts at two northwestern Mediterranean coastal sites, and identified the key physicochemical factors for their occurrence in water using a multivariate analysis. Total viable and culturable cell counts showed the same temporal pattern during the warmer season, whereas the ratios between both methods were inverted during the colder seasons (<15°C), indicating that some of the vibrio community had entered into a viable but non-culturable (VBNC) state. We confirmed that Seawater Surface Temperature explained 51-62% of the total variance in culturable counts, and also showed that the occurrence of viable vibrios is controlled by two variables, pheopigment (15%) and phosphate (12%) concentrations, suggesting that other unidentified factors play a role in maintaining viability.

Cellular bone matrices: viable stem cell-containing bone graft substitutes.

Science.gov (United States)

Skovrlj, Branko; Guzman, Javier Z; Al Maaieh, Motasem; Cho, Samuel K; Iatridis, James C; Qureshi, Sheeraz A

2014-11-01

Advances in the field of stem cell technology have stimulated the development and increased use of allogenic bone grafts containing live mesenchymal stem cells (MSCs), also known as cellular bone matrices (CBMs). It is estimated that CBMs comprise greater than 17% of all bone grafts and bone graft substitutes used. To critically evaluate CBMs, specifically their technical specifications, existing published data supporting their use, US Food and Drug Administration (FDA) regulation, cost, potential pitfalls, and other aspects pertaining to their use. Areview of literature. A series of Ovid, Medline, and Pubmed-National Library of Medicine/National Institutes of Health (www.ncbi.nlm.nih.gov) searches were performed. Only articles in English journals or published with English language translations were included. Level of evidence of the selected articles was assessed. Specific technical information on each CBM was obtained by direct communication from the companies marketing the individual products. Five different CBMs are currently available for use in spinal fusion surgery. There is a wide variation between the products with regard to the average donor age at harvest, total cellular concentration, percentage of MSCs, shelf life, and cell viability after defrosting. Three retrospective studies evaluating CBMs and fusion have shown fusion rates ranging from 90.2% to 92.3%, and multiple industry-sponsored trials are underway. No independent studies evaluating spinal fusion rates with the use of CBMs exist. All the commercially available CBMs claim to meet the FDA criteria under Section 361, 21 CFR Part 1271, and are not undergoing FDA premarket review. The CBMs claim to provide viable MSCs and are offered at a premium cost. Numerous challenges exist in regard to MSCs' survival, function, osteoblastic potential, and cytokine production once implanted into the intended host. Cellular bone matrices may be a promising bone augmentation technology in spinal fusion surgery

Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR (PMA-qPCR).

Science.gov (United States)

Li, Dan; Tong, Tiezheng; Zeng, Siyu; Lin, Yiwen; Wu, Shuxu; He, Miao

2014-02-01

The detection of viable bacteria in wastewater treatment plants (WWTPs) is very important for public health, as WWTPs are a medium with a high potential for waterborne disease transmission. The aim of this study was to use propidium monoazide (PMA) combined with the quantitative polymerase chain reaction (PMA-qPCR) to selectively detect and quantify viable bacteria cells in full-scale WWTPs in China. PMA was added to the concentrated WWTP samples at a final concentration of 100 micromol/L and the samples were incubated in the dark for 5 min, and then lighted for 4 min prior to DNA extraction and qPCR with specific primers for Escherichia coli and Enterococci, respectively. The results showed that PMA treatment removed more than 99% of DNA from non-viable cells in all the WWTP samples, while matrices in sludge samples markedly reduced the effectiveness of PMA treatment. Compared to qPCR, PMA-qPCR results were similar and highly linearly correlated to those obtained by culture assay, indicating that DNA from non-viable cells present in WWTP samples can be eliminated by PMA treatment, and that PMA-qPCR is a reliable method for detection of viable bacteria in environmental samples. This study demonstrated that PMA-qPCR is a rapid and selective detection method for viable bacteria in WWTP samples, and that WWTPs have an obvious function in removing both viable and non-viable bacteria. The results proved that PMA-qPCR is a promising detection method that has a high potential for application as a complementary method to the standard culture-based method in the future.

Evaluating the interacting influences of pollination, seed predation, invasive species and isolation on reproductive success in a threatened alpine plant.

Science.gov (United States)

Krushelnycky, Paul D

2014-01-01

Reproduction in rare plants may be influenced and limited by a complex combination of factors. External threats such as invasive species and landscape characteristics such as isolation may impinge on both pollination and seed predation dynamics, which in turn can strongly affect reproduction. I assessed how patterns in floral visitation, seed predation, invasive ant presence, and plant isolation influenced one another and ultimately affected viable seed production in Haleakalā silverswords (Argyroxiphium sandwicense subsp. macrocephalum) of Hawai'i. Floral visitation was dominated by endemic Hylaeus bees, and patterns of visitation were influenced by floral display size and number of plants clustered together, but not by floral herbivory or nearest flowering neighbor distance. There was also some indication that Argentine ant presence impacted floral visitation, but contradictory evidence and limitations of the study design make this result uncertain. Degree of seed predation was associated only with plant isolation, with the two main herbivores partitioning resources such that one preferentially attacked isolated plants while the other attacked clumped plants; total seed predation was greater in more isolated plants. Net viable seed production was highly variable among individuals (0-55% seed set), and was affected mainly by nearest neighbor distance, apparently owing to low cross-pollination among plants separated by even short distances (>10-20 m). This isolation effect dominated net seed set, with no apparent influence from floral visitation rates, percent seed predation, or invasive ant presence. The measured steep decline in seed set with isolation distance may not be typical of the entire silversword range, and may indicate that pollinators in addition to Hylaeus bees could be important for greater gene flow. Management aimed at maintaining or maximizing silversword reproduction should focus on the spatial context of field populations and outplanting

Excimer laser forward transfer of mammalian cells using a novel triazene absorbing layer

International Nuclear Information System (INIS)

Doraiswamy, A.; Narayan, R.J.; Lippert, T.; Urech, L.; Wokaun, A.; Nagel, M.; Hopp, B.; Dinescu, M.; Modi, R.; Auyeung, R.C.Y.; Chrisey, D.B.

2006-01-01

We present a novel laser-based approach for developing tissue engineered constructs and other cell-based assembly's. We have deposited mesoscopic patterns of viable B35 neuroblasts using a soft direct approach of the matrix assisted pulsed laser evaporation direct write (MAPLE DW) process. As a development of the conventional direct write process, an intermediate layer of absorbing triazene polymer is used to provide gentler and efficient transfers. Transferred cells were examined for viability and proliferation and compared with that of as-seeded cells to determine the efficacy of the process. Results suggest that successful transfers can be achieved at lower fluences than usual by the incorporation of the intermediate absorbing layer thus avoiding any damage to cells and other delicate materials. MAPLE DW offers rapid computer-controlled deposition of mesoscopic voxels at high spatial resolutions, with extreme versatility in depositing combinations of natural/synthetic, living/non-living, organic/inorganic and hard/soft materials. Our approach offers a gentle and efficient transfer of viable cells which when combined with a variety of matrix materials allows development of constructs and bioactive systems in bioengineering

Viable group A streptococci in macrophages during acute soft tissue infection.

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Pontus Thulin

2006-03-01

Full Text Available Group A streptococcal severe soft tissue infections, such as necrotizing fasciitis, are rapidly progressive infections associated with high mortality. Group A streptococcus is typically considered an extracellular pathogen, but has been shown to reside intracellularly in host cells.We characterized in vivo interactions between group A streptococci (GAS and cells involved in innate immune responses, using human biopsies (n = 70 collected from 17 patients with soft tissue infections. Immunostaining and in situ image analysis revealed high amounts of bacteria in the biopsies, even in those collected after prolonged antibiotic therapy. Viability of the streptococci was assessed by use of a bacterial viability stain, which demonstrated viable bacteria in 74% of the biopsies. GAS were present both extracellularly and intracellularly within phagocytic cells, primarily within macrophages. Intracellular GAS were predominantly noted in biopsies from newly involved tissue characterized by lower inflammation and bacterial load, whereas purely extracellular GAS or a combination of intra- and extracellular GAS dominated in severely inflamed tissue. The latter tissue was also associated with a significantly increased amount of the cysteine protease streptococcal pyrogenic exotoxin SpeB. In vitro studies confirmed that macrophages serve as reservoirs for viable GAS, and infection with a speB-deletion mutant produced significantly lower frequencies of cells with viable GAS following infection as compared to the wild-type bacteria.This is the first study to demonstrate that GAS survive intracellularly in macrophages during acute invasive infections. This intracellular presence may have evolved as a mechanism to avoid antibiotic eradication, which may explain our finding that high bacterial load is present even in tissue collected after prolonged intravenous antibiotic therapy. This new insight into the pathogenesis of streptococcal soft tissue infections

Viable Group A Streptococci in Macrophages during Acute Soft Tissue Infection.

Directory of Open Access Journals (Sweden)

2006-01-01

Full Text Available BACKGROUND: Group A streptococcal severe soft tissue infections, such as necrotizing fasciitis, are rapidly progressive infections associated with high mortality. Group A streptococcus is typically considered an extracellular pathogen, but has been shown to reside intracellularly in host cells. METHODS AND FINDINGS: We characterized in vivo interactions between group A streptococci (GAS and cells involved in innate immune responses, using human biopsies (n = 70 collected from 17 patients with soft tissue infections. Immunostaining and in situ image analysis revealed high amounts of bacteria in the biopsies, even in those collected after prolonged antibiotic therapy. Viability of the streptococci was assessed by use of a bacterial viability stain, which demonstrated viable bacteria in 74% of the biopsies. GAS were present both extracellularly and intracellularly within phagocytic cells, primarily within macrophages. Intracellular GAS were predominantly noted in biopsies from newly involved tissue characterized by lower inflammation and bacterial load, whereas purely extracellular GAS or a combination of intra- and extracellular GAS dominated in severely inflamed tissue. The latter tissue was also associated with a significantly increased amount of the cysteine protease streptococcal pyrogenic exotoxin SpeB. In vitro studies confirmed that macrophages serve as reservoirs for viable GAS, and infection with a speB-deletion mutant produced significantly lower frequencies of cells with viable GAS following infection as compared to the wild-type bacteria. CONCLUSIONS: This is the first study to demonstrate that GAS survive intracellularly in macrophages during acute invasive infections. This intracellular presence may have evolved as a mechanism to avoid antibiotic eradication, which may explain our finding that high bacterial load is present even in tissue collected after prolonged intravenous antibiotic therapy. This new insight into the pathogenesis

Depth of dormancy in tomato (Lycopersicon esculentum Mill.) seeds is related to the progression of the cell cycle prior to the induction of dormancy

NARCIS (Netherlands)

Castro, de R.D.; Bino, R.J.; Jing, H.C.; Hilhorst, H.W.M.

2001-01-01

Cell cycle activities are initiated following imbibition of non-dormant seeds. However, it is not known whether cell cycle related events other than DNA replication also remain suppressed in imbibed dormant seeds. The objective of this study was to demonstrate that the transitions between the

Polyelectrolyte Complex Beads by Novel Two-Step Process for Improved Performance of Viable Whole-Cell Baeyer-Villiger Monoxygenase by Immobilization

Czech Academy of Sciences Publication Activity Database

Krajčovič, T.; Bučko, M.; Vikartovská, A.; Lacík, I.; Uhelská, L.; Chorvát, D.; Neděla, Vilém; Tihlaříková, Eva; Gericke, M.; Heinze, T.; Gemeiner, P.

2017-01-01

Roč. 7, č. 11 (2017), s. 353-364 ISSN 2073-4344 Institutional support: RVO:68081731 Keywords : polyelectrolyte complex beads * environmental scanning electron microscopy * confocal laser scanning microscopy * Baeyer-Villiger biooxidation * cyclohexanone monoxygenase * immobilization * viable whole-cell biocatalyst Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering OBOR OECD: Bioprocessing technologies (industrial processes relying on biological agents to drive the process) biocatalysis, fermentation Impact factor: 3.082, year: 2016 http://www.mdpi.com/2073-4344/7/11/353

Viable suspensions of maize (Zea mays L.) pollen with exogenous DNA

Energy Technology Data Exchange (ETDEWEB)

Broglia, M. [ENEA, Centro Ricerche Casaccia, Rome (Italy). Dip. Innovazione

1996-12-01

A viable suspension of maize pollen in aqueous medium containing exogenous DNA would be a suitable tool in attempting maize genetic transformation via pollen grains by different techniques. In this work the effects of addition of DNA to hypertonic aqueous media able to preserve maize pollen viability were investigated. An almost total loss of viability was found when pollen was incubated with native DNA in water or sucrose medium due to the immediate sticking of DNA on the pollen wall. Calcium in the incubation medium avoided DNA sticking preserving pollen fertilization ability. Pre-washing of pollen in hypertonic sucrose solution was proved to remove DNA binding components from the pollen wall. PEG 20%, that is known to inhibit pollen, and silk nucleases, was also used instead of sucrose, without any reduction in the seed-set yields.

Sunflower seed for human consumption as a substrate for the growth of mycopopulations

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Škrinjar Marija M.

2012-01-01

Full Text Available These mycological investigations are implicating samples of protein sunflower seed from regular cultivation in the Vojvodina Province. Samples are examined in different stages of production: reception in the silo, separation of massive fraction on peeler and then peeling, kernel after peeling, hull, final product, i.e. kernels separated from visible impurities on conveyor bel, that are later manually divided in two fractions - a seemingly whole, undamaged kernels, without change of colour, and b seemingly damaged kernels, broken, with change of colour. For the determination of viable count of moulds and their isolation, two different media are used in parallel: Sabouraud maltose agar (SMA and malt/yeast extract with 50% of glucose (MY50G, favourable for growth of xerophilic moulds. All samples tested were contaminated with fungi. Total viable mould count per seed varied from 1.6 (SMA respecting 1.3 (MY50G on reception, to 5.6 (SMA and 7.5 (MY50G cfu/seed in visually damaged sunflower kernels (final product. From seeds, kernels and hull, numerous moulds were isolated, belonging to 8 genera and 13 species (Alternaria alternata, Arthrinium phaeospermum, Aspergillus candidus, A. flavus, A. niger, A. ochraceus, A. versicolor, A. wentii, Cladosporium cladosporioides, Eurotium herbariorum, Penicillium aurantiogriseum, Rhizopus stolonifer and Trichoderma harzianum. Alternaria alternata, Aspergillus flavus, A.ochraceus, A. versicolor and Eurotium herbariorum were isolated on both media. Aspergillus candidus, A. versicolor, C. Cladosporioides, P. aurantiogriseum and T. harzianum were isolated only on SMA, while A. niger, A. wentii and R. stolonifer were exclusively isolated on MY50G. Most ubiquitous species is A. alternata, which is isolated from all tested samples, while A. candidus, C. cladosporioides and T. harzianum were isolated from sunflower seed on reception in silo, using SMA medium.

The biocompatibility of titanium cardiovascular devices seeded with autologous blood-derived endothelial progenitor cells: EPC-seeded antithrombotic Ti implants.

Science.gov (United States)

Achneck, Hardean E; Jamiolkowski, Ryan M; Jantzen, Alexandra E; Haseltine, Justin M; Lane, Whitney O; Huang, Jessica K; Galinat, Lauren J; Serpe, Michael J; Lin, Fu-Hsiung; Li, Madison; Parikh, Amar; Ma, Liqiao; Chen, Tao; Sileshi, Bantayehu; Milano, Carmelo A; Wallace, Charles S; Stabler, Thomas V; Allen, Jason D; Truskey, George A; Lawson, Jeffrey H

2011-01-01

Implantable and extracorporeal cardiovascular devices are commonly made from titanium (Ti) (e.g. Ti-coated Nitinol stents and mechanical circulatory assist devices). Endothelializing the blood-contacting Ti surfaces of these devices would provide them with an antithrombogenic coating that mimics the native lining of blood vessels and the heart. We evaluated the viability and adherence of peripheral blood-derived porcine endothelial progenitor cells (EPCs), seeded onto thin Ti layers on glass slides under static conditions and after exposure to fluid shear stresses. EPCs attached and grew to confluence on Ti in serum-free medium, without preadsorption of proteins. After attachment to Ti for 15 min, less than 5% of the cells detached at a shear stress of 100 dyne / cm(2). Confluent monolayers of EPCs on smooth Ti surfaces (Rq of 10 nm), exposed to 15 or 100 dyne/cm(2) for 48 h, aligned and elongated in the direction of flow and produced nitric oxide dependent on the level of shear stress. EPC-coated Ti surfaces had dramatically reduced platelet adhesion when compared to uncoated Ti surfaces. These results indicate that peripheral blood-derived EPCs adhere and function normally on Ti surfaces. Therefore EPCs may be used to seed cardiovascular devices prior to implantation to ameliorate platelet activation and thrombus formation. Copyright © 2010 Elsevier Ltd. All rights reserved.

Detection and correction of patient movement in prostate brachytherapy seed reconstruction

Science.gov (United States)

Lam, Steve T.; Cho, Paul S.; Marks, Robert J., II; Narayanan, Sreeram

2005-05-01

Intraoperative dosimetry of prostate brachytherapy can help optimize the dose distribution and potentially improve clinical outcome. Evaluation of dose distribution during the seed implant procedure requires the knowledge of 3D seed coordinates. Fluoroscopy-based seed localization is a viable option. From three x-ray projections obtained at different gantry angles, 3D seed positions can be determined. However, when local anaesthesia is used for prostate brachytherapy, the patient movement during fluoroscopy image capture becomes a practical problem. If uncorrected, the errors introduced by patient motion between image captures would cause seed mismatches. Subsequently, the seed reconstruction algorithm would either fail to reconstruct or yield erroneous results. We have developed an algorithm that permits detection and correction of patient movement that may occur between fluoroscopy image captures. The patient movement is decomposed into translational shifts along the tabletop and rotation about an axis perpendicular to the tabletop. The property of spatial invariance of the co-planar imaging geometry is used for lateral movement correction. Cranio-caudal movement is corrected by analysing the perspective invariance along the x-ray axis. Rotation is estimated by an iterative method. The method can detect and correct for the range of patient movement commonly seen in the clinical environment. The algorithm has been implemented for routine clinical use as the preprocessing step for seed reconstruction.

Detection of viable Escherichia coli O157:H7 in ground beef by propidium monoazide real-time PCR.

Science.gov (United States)

Liu, Yarui; Mustapha, Azlin

2014-01-17

Escherichia coli O157:H7 associated with food has caused many serious public health problems in recent years. However, only viable cells of this pathogen can cause infections, and false-positive detection caused by dead cells can lead to unnecessary product recalls. The objective of this study was to develop and optimize a method that combines propidium monoazide (PMA) staining with real-time PCR to detect only viable cells of E. coli O157:H7 in ground beef. PMA is a DNA intercalating dye that can penetrate compromised membranes of dead cells and bind to cellular DNA, preventing its amplification via a subsequent PCR. Three strains of E. coli O157:H7 (505B, G5310 and C7927) at concentrations of 10(0) to 10(8)CFU/mL were used as live cells. Dead cells were obtained by heating cell suspensions at 85°C for 15 min. Suspensions were treated with PMA and the optimized assay was applied to artificially contaminated ground beef with two different fat contents (10% and 27%). DNA was extracted and amplified by TaqMan® real-time PCR assay targeting the uidA gene for detection of E. coli O157:H7. Plasmid pUC19 was added as an internal amplification control (IAC). A treatment of 25 μM PMA with a 10-min light exposure on ice was sufficient to eliminate DNA from 10(8) dead E. coli O157:H7 cells/mL. The optimized assay could detect as low as 10(2) CFU/mL viable E. coli O157:H7 in pure culture and 10(5) CFU/g in ground beef, in the presence of 10(6)/mL or g of dead cells. With an 8-h enrichment, 1 CFU/g viable E. coli O157:H7 in ground beef was detectable without interference from 10(6) dead cells/g. In conclusion, the PMA real-time PCR could effectively detect viable E. coli O157:H7 without being compromised by dead cells. Copyright © 2013 Elsevier B.V. All rights reserved.

In vitro inhibition of Eimeria tenella sporozoite invasion into host cells by probiotics.

Science.gov (United States)

Hessenberger, S; Schatzmayr, G; Teichmann, K

2016-10-15

The aim was to study the effects of probiotics isolated from the intestinal tract of livestock animals on Eimeria tenella invasion into Madin-Darby bovine kidney (MDBK) cells in vitro. E. tenella sporozoites were purified and labeled with 5(6)-carboxyfluorescein diacetate N-succinimidyl ester before seeding on cell cultures, and invasion was evaluated by fluorescence microscopy. Two protocols (A and B) were used. In protocol A, Enterococcus faecium # 589 or Lactobacillus salivarius subsp. salivarius # 505 were added together with sporozoites to MDBK cell cultures and invasion was evaluated after incubation for approximately 20h. Viable, dead, or spent culture supernatants of probiotics were tested. In protocol B, viable probiotics were incubated with MDBK cells for one hour before sporozoites were added and invasion was evaluated after two more hours of incubation. Parasite invasion of viable, dead, or spent culture supernatant of E. faecium # 589 was assessed. Using protocol A, it was shown that parasite invasion was inhibited by viable (80%) or dead (75%) E. faecium # 589. While inhibition by viable L. salivarius subsp. salivarius # 505 was not valid at the highest concentration and not significant at the other test concentrations, dead cells inhibited parasite invasion up to 45%. Spent culture supernatants of both probiotics had no influence on parasite invasion. Using protocol B, it was shown that viable Bifidobacterium animalis subsp. animalis # 503, E. faecium # 497, E. faecium # 589, L. reuteri # 514, L. salivarius subsp. salivarius # 505, and Bacillus subtilis # 588 inhibited parasite invasion into MDBK cells up to 80%. Anticoccidial activity was strain-specific for E. faecium strains, and the strongest effect was shown by E. faecium # 589. Anticoccidial effects of some of the tested probiotics have already been shown in vivo, which makes them candidates to prevent coccidiosis. These findings have now been confirmed in vitro. The used parasite invasion

Effect of Time and Burial Depth on Breaking Seed dormancy and Germination of Weed Seeds

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marzie mazhari

2016-02-01

Full Text Available Introduction: Weeds limit crop growth, development and yield through competing. Seed bank of weeds in field is one of the sources which can affect weed management and their control methods. Environmental conditions during seed maturation and following dispersal interact to influence the germination phenology of many species. Disturbance plays a key role in the maintenance of habitat for many plant species, particularly referrals, for example, fire ephemerals, desert annuals, and arable weeds. Seed germination and emergence depend on endogenous and exogenous factors. Viable seeds are dormant when all environmental conditions are appropriate for germination but seeds fail to germinate. Thus, dormancy plays an important ecological role in preventing seed germination, being a major contributor to seed persistence of some species in soil. Buried seeds of annual weeds are certainly subjected to different soil moisture conditions during their dormancy release season (winter according to the annual rainfall pattern and burial depth. Shallow buried seeds are exposed to soil moisture fluctuations that could affect their dormancy status. Laboratory studies showed that desiccation and subsequent re-hydration of seeds could stimulate germination and modify seed light requirements. Seeds buried in deeper layers of the soil would not be exposed to such fluctuations in soil moisture, but would be exposed to different soil moisture environments depending on weather and soil characteristics. The effects of interactions between temperature, and soil or seed moisture, on seed dormancy changes have been reported for several species. Therefore, the objectives of this study were to determine the effect of time and burial depth treatments on seed germination and seedling emergence of Aegilops cylindrica, Agropyrom repens, Avena fatua, Bromus dantoniae, Cynodon dactylon, Cyprus rotundus, Setaria viridis, Anthriscus sylvestris, Centurea cyanus. Materials and Methods: In

Toxicity assessment and modelling of Moringa oleifera seeds in water purification by whole cell bioreporter.

Science.gov (United States)

Al-Anizi, Ali Adnan; Hellyer, Maria Theresa; Zhang, Dayi

2014-06-01

Moringa oleifera has been used as a coagulation reagent for drinking water purification, especially in developing countries such as Malawi. This research revealed the cytoxicity and genotoxicity of M. oleifera by Acinetobacter bioreporter. The results indicated that significant cytoxicity effects were observed when the powdered M. oleifera seeds concentration is from 1 to 50 mg/L. Through direct contact, ethanolic-water extraction and hexane extraction, the toxic effects of hydrophobic and hydrophilic components in M. oleifera seeds were distinguished. It suggested that the hydrophobic lipids contributed to the dominant cytoxicity, consequently resulting in the dominant genotoxicity in the water-soluble fraction due to limited dissolution when the M. oleifera seeds granule concentration was from 10 to 1000 mg/L. Based on cytoxicity and genotoxicity model, the LC50 and LC90 of M. oleifera seeds were 8.5 mg/L and 300 mg/L respectively and their genotoxicity was equivalent to 8.3 mg mitomycin C per 1.0 g dry M. oleifera seed. The toxicity of M. oleifera has also remarkable synergistic effects, suggesting whole cell bioreporter as an appropriate and complementary tool to chemical analysis for environmental toxicity assessment. Copyright © 2014 Elsevier Ltd. All rights reserved.

Running title: Water distribution in chickpea seeds

African Journals Online (AJOL)

agriphy20

2012-07-24

Jul 24, 2012 ... molecular mobility of cellular water in magnetically exposed seeds as compared to unexposed seeds. Analysis of ... protrusion takes place through the seed coat and absorption .... directly related to water activity (aw) of the cell water. (Gambhir ..... plants, including photosynthesis, respiration and enzymatic ...

Viable bacteria associated with red blood cells and plasma in freshly drawn blood donations.

Science.gov (United States)

Damgaard, Christian; Magnussen, Karin; Enevold, Christian; Nilsson, Martin; Tolker-Nielsen, Tim; Holmstrup, Palle; Nielsen, Claus Henrik

2015-01-01

Infection remains a leading cause of post-transfusion mortality and morbidity. Bacterial contamination is, however, detected in less than 0.1% of blood units tested. The aim of the study was to identify viable bacteria in standard blood-pack units, with particular focus on bacteria from the oral cavity, and to determine the distribution of bacteria revealed in plasma and in the red blood cell (RBC)-fraction. Cross-sectional study. Blood were separated into plasma and RBC-suspensions, which were incubated anaerobically or aerobically for 7 days on trypticase soy blood agar (TSA) or blue lactose plates. For identification colony PCR was performed using primers targeting 16S rDNA. Blood donors attending Capital Region Blood Bank, Copenhagen University Hospital, Rigshospitalet, Hvidovre, Denmark, October 29th to December 10th 2013. 60 donors (≥50 years old), self-reported medically healthy. Bacterial growth was observed on plates inoculated with plasma or RBCs from 62% of the blood donations. Growth was evident in 21 (35%) of 60 RBC-fractions and in 32 (53%) of 60 plasma-fractions versus 8 of 60 negative controls (p = 0.005 and p = 2.6x10-6, respectively). Propionibacterium acnes was found in 23% of the donations, and Staphylococcus epidermidis in 38%. The majority of bacteria identified in the present study were either facultative anaerobic (59.5%) or anaerobic (27.8%) species, which are not likely to be detected during current routine screening. Viable bacteria are present in blood from donors self-reported as medically healthy, indicating that conventional test systems employed by blood banks insufficiently detect bacteria in plasma. Further investigation is needed to determine whether routine testing for anaerobic bacteria and testing of RBC-fractions for adherent bacteria should be recommended.

GROWTH OF A LOCALIZED SEED MAGNETIC FIELD IN A TURBULENT MEDIUM

International Nuclear Information System (INIS)

Cho, Jungyeon; Yoo, Hyunju

2012-01-01

Turbulence dynamo deals with the amplification of a seed magnetic field in a turbulent medium and has been studied mostly for uniform or spatially homogeneous seed magnetic fields. However, some astrophysical processes (e.g., jets from active galaxies, galactic winds, or ram-pressure stripping in galaxy clusters) can provide localized seed magnetic fields. In this paper, we numerically study amplification of localized seed magnetic fields in a turbulent medium. Throughout the paper, we assume that the driving scale of turbulence is comparable to the size of the system. Our findings are as follows. First, turbulence can amplify a localized seed magnetic field very efficiently. The growth rate of magnetic energy density is as high as that for a uniform seed magnetic field. This result implies that magnetic field ejected from an astrophysical object can be a viable source of a magnetic field in a cluster. Second, the localized seed magnetic field disperses and fills the whole system very fast. If turbulence in a system (e.g., a galaxy cluster or a filament) is driven at large scales, we expect that it takes a few large-eddy turnover times for the magnetic field to fill the whole system. Third, growth and turbulence diffusion of a localized seed magnetic field are also fast in high magnetic Prandtl number turbulence. Fourth, even in decaying turbulence, a localized seed magnetic field can ultimately fill the whole system. Although the dispersal rate of the magnetic field is not fast in purely decaying turbulence, it can be enhanced by an additional forcing.

Study on Seed Morphogenesis of Orobanchaceae in Taiwan

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Jao-Shien Chen

2011-11-01

Full Text Available Seed morphogenesis of Orobanchaceae was not completely investigated previously. Here, we observed seed development of Orobanchaceous species in Taiwan using light and scanning electron microscopies. Results indicated that seeds of Aeginetia indica, Boschniakia himalaica, and Orobanche caerulescens all consisted of embryo, endosperm and testa. Ontogeny of the embryo in A. indica was Solanad type, while in both B. himalaica and O. caerulescens was Onagrad type. The mature embryos of the three species lacked embryonic organs, and their endosperm development was the cellular type and, at maturity, appeared as several cell layers of storage tissue. Ontogeny of the testa was all non-multiplicative, with the residues of the outermost cell layer and reticulately-thickened secondary walls of its cells at maturity. Mature seeds of A. indica and O. caerulescens were ovate whereas those of B. himalaica were oblate. As for Christisonia hookeri, due to lack of samples, only the cellular-typed endosperm was determined. The comparative development of Orobanchaceous seeds was discussed.

Droplet digital PCR improves absolute quantification of viable lactic acid bacteria in faecal samples.

Science.gov (United States)

Gobert, Guillaume; Cotillard, Aurélie; Fourmestraux, Candice; Pruvost, Laurence; Miguet, Jean; Boyer, Mickaël

2018-03-14

Analysing correlations between the observed health effects of ingested probiotics and their survival in digestive tract allows adapting their preparations for food. Tracking ingested probiotic in faecal samples requires accurate and specific tools to quantify live vs dead cells at strain level. Traditional culture-based methods are simpler to use but they do not allow quantifying viable but non-cultivable (VBNC) cells and they are poorly discriminant below the species level. We have set up a viable PCR (vPCR) assay combining propidium monoazide (PMA) treatment and either real time quantitative PCR (qPCR) or droplet digital PCR (ddPCR) to quantify a Lactobacillus rhamnosus and two Lactobacillus paracasei subsp. paracasei strains in piglet faeces. Adjustments of the PMA treatment conditions and reduction of the faecal sample size were necessary to obtain accurate discrimination between dead and live cells. The study also revealed differences of PMA efficiency among the two L. paracasei strains. Both PCR methods were able to specifically quantify each strain and provided comparable total bacterial counts. However, quantification of lower numbers of viable cells was best achieved with ddPCR, which was characterized by a reduced lower limit of quantification (improvement of up to 1.76 log 10 compared to qPCR). All three strains were able to survive in the piglets' gut with viability losses between 0.78 and 1.59 log 10 /g faeces. This study shows the applicability of PMA-ddPCR to specific quantification of small numbers of viable bacterial cells in the presence of an important background of unwanted microorganisms, and without the need to set up standard curves. It also illustrates the need to adapt PMA protocols according to the final matrix and target strain, even for closely related strains. The PMA-ddPCR approach provides a new tool to quantify bacterial survival in faecal samples from a preclinical and clinical trial. Copyright © 2018 The Authors. Published by

Utilization of deoiled Jatropha curcas seed cake for production of xylanase from thermophilic Scytalidium thermophilum.

Science.gov (United States)

Joshi, Chetna; Khare, S K

2011-01-01

Jatropha curcas is a major biodiesel crop. Large amount of deoiled cake is generated as by-product during biodiesel production from its seeds. Deoiled J. curcas seed cake was assessed as substrate for the production of xylanase from thermophilic fungus Scytalidium thermophilum by solid-state fermentation. The seed cake was efficiently utilized by S. thermophilum for its growth during which it produced good amount of heat stable extracellular xylanase. The solid-state fermentation conditions were optimized for maximum xylanase production. Under the optimized conditions viz. deoiled seed cake supplemented with 1% oat-spelt xylan, adjusted to pH 9.0, moisture content 1:3 w/v, inoculated with 1×10(6) spores per 5 g cake and incubated at 45 °C, 1455 U xylanase/g deoiled seed cake was obtained. The xylanase was useful in biobleaching of paper pulp. Solid-state fermentation of deoiled cake appears a potentially viable approach for its effective utilization. Copyright © 2010 Elsevier Ltd. All rights reserved.

The Arabidopsis GASA10 gene encodes a cell wall protein strongly expressed in developing anthers and seeds.

Science.gov (United States)

Trapalis, Menelaos; Li, Song Feng; Parish, Roger W

2017-07-01

The Arabidopsis GASA10 gene encodes a GAST1-like (Gibberellic Acid-Stimulated) protein. Reporter gene analysis identified consistent expression in anthers and seeds. In anthers expression was developmentally regulated, first appearing at stage 7 of anther development and reaching a maximum at stage 11. Strongest expression was in the tapetum and developing microspores. GASA10 expression also occurred throughout the seed and in root vasculature. GASA10 was shown to be transported to the cell wall. Using GASA1 and GASA6 as positive controls, gibberellic acid was found not to induce GASA10 expression in Arabidopsis suspension cells. Overexpression of GASA10 (35S promoter-driven) resulted in a reduction in silique elongation. GASA10 shares structural similarities to the antimicrobial peptide snakin1, however, purified GASA10 failed to influence the growth of a variety of bacterial and fungal species tested. We propose cell wall associated GASA proteins are involved in regulating the hydroxyl radical levels at specific sites in the cell wall to facilitate wall growth (regulating cell wall elongation). Copyright © 2017 Elsevier B.V. All rights reserved.

Internal seed dispersal by parrots: an overview of a neglected mutualism

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Guillermo Blanco

2016-02-01

Full Text Available Despite the fact that parrots (Psitacifformes are generalist apex frugivores, they have largely been considered plant antagonists and thus neglected as seed dispersers of their food plants. Internal dispersal was investigated by searching for seeds in faeces opportunistically collected at communal roosts, foraging sites and nests of eleven parrot species in different habitats and biomes in the Neotropics. Multiple intact seeds of seven plant species of five families were found in a variable proportion of faeces from four parrot species. The mean number of seeds of each plant species per dropping ranged between one and about sixty, with a maximum of almost five hundred seeds from the cacti Pilosocereus pachycladus in a single dropping of Lear’s Macaw (Anodorhynchus leari. All seeds retrieved were small (<3 mm and corresponded to herbs and relatively large, multiple-seeded fleshy berries and infrutescences from shrubs, trees and columnar cacti, often also dispersed by stomatochory. An overview of the potential constraints driving seed dispersal suggest that, despite the obvious size difference between seeds dispersed by endozoochory and stomatochory, there is no clear difference in fruit size depending on the dispersal mode. Regardless of the enhanced or limited germination capability after gut transit, a relatively large proportion of cacti seeds frequently found in the faeces of two parrot species were viable according to the tetrazolium test and germination experiments. The conservative results of our exploratory sampling and a literature review clearly indicate that the importance of parrots as endozoochorous dispersers has been largely under-appreciated due to the lack of research systematically searching for seeds in their faeces. We encourage the evaluation of seed dispersal and other mutualistic interactions mediated by parrots before their generalized population declines contribute to the collapse of key ecosystem processes.

Does enterohemorrhagic Escherichia coli O157:H7 enter the viable but nonculturable state in salted salmon roe?

Science.gov (United States)

Makino, S I; Kii, T; Asakura, H; Shirahata, T; Ikeda, T; Takeshi, K; Itoh, K

2000-12-01

An outbreak caused by salted salmon roe contaminated with enterohemorrhagic Escherichia coli O157 occurred in Japan in 1998. Since about 0.75 to 1.5 viable cells were estimated to cause infection, we presumed that O157 might enter the viable but nonculturable (VNC) state in salted salmon roe and consequently that viable cell numbers might be underestimated. Although patient-originating O157 cells could not grow on agar plates after 72 h of incubation in 13% NaCl, they were resuscitated in yeast extract broth, and more than 90% of the cells were shown to be viable by fluorescent staining, suggesting that almost all of them could enter the VNC state in NaCl water. Roe-originating O157 was resistant to NaCl because it could grow on agar after 72 h of incubation in NaCl water, but about 20% of cells appeared to enter the VNC state. Therefore, germfree mice were infected with O157 to examine the resuscitation of cells in the VNC state and the retention of pathogenicity. O157 that originated in roe, but not patients, killed mice and was isolated from the intestine. However, these isolates had become sensitive to NaCl. O157 cells of roe origin incubated in normal media also killed mice and were isolated from the intestine, but they also became transiently NaCl sensitive. We therefore propose that bacterial cells might enter the VNC state under conditions of stress, such as those encountered in vivo or in high salt concentrations, and then revive when those conditions have eased. If so, the VNC state in food is potentially dangerous from a public health viewpoint and may have to be considered at the time of food inspection. Finally, the establishment of a simple recovery system for VNC cells should be established.

[Seed geography: its concept and basic scientific issues].

Science.gov (United States)

Yu, Shun-Li; Wang, Zong-Shuai; Zeren, Wangmu

2010-01-01

In this paper, a new concept 'seed geography' was provided, and its definition, research contents, and scientific issues were put forward. Seed geography is a newly developed interdisciplinary science from plant geography, seed ecology, and phytosociology, which studies the geographic variation patterns of seed biological traits as well as their relationships with environmental factors from macroscopic to microscopic, and the seed formation, development, and change trends. The main research contents would include geography of seed mass, geography of seed chemical components, geography of seed morphology, geography of seed cell biological characteristics, geography of seed physiological characteristics, geography of seed genetic characteristics, and geography of flower and fruit. To explore the scientific issues in seed geography would help us to better understand the long-term adaptation and evolution of seed characteristics to natural environments.

Edema-induced increase in tumour cell survival for 125I and 103Pd prostate permanent seed implants - a bio-mathematical model

International Nuclear Information System (INIS)

Yue Ning; Chen Zhe; Nath, Ravinder

2002-01-01

Edema caused by the surgical procedure of prostate seed implantation expands the source-to-point distances within the prostate and hence decreases the dose coverage. The decrease of dose coverage results in an increase in tumour cell survival. To investigate the effects of edema on tumour cell survival, a bio-mathematical model of edema and the corresponding cell killing by continuous low dose rate irradiation (CLDRI) was developed so that tumour cell surviving fractions can be estimated in an edematous prostate for both 125 I and 103 Pd seed implants. The dynamic nature of edema and its resolution were modelled with an exponential function V(T)=V p (1+M exp(-0.693T/T e )) where V p is the prostate volume before implantation, M is the edema magnitude and T e is edema half-life (EHL). The dose rate of a radioactive seed was calculated according to AAPM TG43, i.e. D radical S k Δg(r) φ-bar an /r 2 , where r is the distance between a seed and a given point. The distance r is now a function of time because of edema. The g(r) was approximated as 1/r 0.4 and 1/r 0.8 for 125 I and 103 Pd, respectively. By expanding the mathematical expression of the resultant dose rate in a Taylor series of exponential functions of time, the dose rate was made equivalent to that produced from multiple fictitious radionuclides of different decay constants and strengths. The biologically effective dose (BED) for an edematous prostate implant was then calculated using a generalized Dale equation. The cell surviving fraction was computed as exp(-αBED), where α is the linear coefficient of the survival curve. The tumour cell survival was calculated for both 125 I and 103 Pd seed implants and for different tumour potential doubling time (TPDT) (from 5 days to 30 days) and for edemas of different magnitudes (from 0% to 95%) and edema half-lives (from 4 days to 30 days). Tumour cell survival increased with the increase of edema magnitude and EHL. For a typical edema of a half-life of 10 days

Evaluation of chemopreventive and cytotoxic effect of lemon seed extracts on human breast cancer (MCF-7) cells.

Science.gov (United States)

Kim, Jinhee; Jayaprakasha, Guddadarangavvanahally K; Uckoo, Ram M; Patil, Bhimanagouda S

2012-02-01

Extracts from lemon seed were investigated for the radical scavenging activity and apoptotic effects in human breast adenocarcinoma (MCF-7) cells and non-malignant breast (MCF-12F) cells for the first time. Defatted seed powder was successively extracted with ethyl acetate (EtOAc), acetone, methanol (MeOH), and MeOH:water (80:20). The chemical constituents were identified and quantified by LC-MS and HPLC analysis, respectively. The highest radical scavenging activity of 62.2% and 91.3% was exhibited by MeOH:water (80:20) at 833μg/mL in 1,1-diphenyl-2-picryl hydrazyl (DPPH) and 2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS(+)), respectively. In addition, the MeOH:water (80:20) extract showed the highest (29.1%, Pwater (80:20) extract induced DNA fragmentation and poly(ADP-ribose) polymerase (PARP) cleavage. Increased levels of Bax and cytosolic cytochrome C and decreased levels of Bcl2 were also observed in MeOH:water (80:20) treated MCF-7 cells. In conclusion, the MeOH:water (80:20) extract from lemon seed has potent antioxidant activity and induces apoptosis in MCF-7 cells, leading to the inhibition of proliferation. These results suggest that aglycones and glucosides of the limonoids and flavonoid present in MeOH:water (80:20) extract may potentially serve as a chemopreventive agent for breast cancer. Copyright © 2011 Elsevier Ltd. All rights reserved.

Role of Blossoms in Watermelon Seed Infestation by Acidovorax avenae subsp. citrulli.

Science.gov (United States)

Walcott, R R; Gitaitis, R D; Castro, A C

2003-05-01

ABSTRACT The role of watermelon blossom inoculation in seed infestation by Acidovorax avenae subsp. citrulli was investigated. Approximately 98% (84/87) of fruit developed from blossoms inoculated with 1 x 10(7) or 1 x 10(9) CFU of A. avenae subsp. citrulli per blossom were asymptomatic. Using immunomagnetic separation and the polymerase chain reaction, A. avenae subsp. citrulli was detected in 44% of the seed lots assayed, despite the lack of fruit symptoms. Furthermore, viable colonies were recovered from 31% of the seed lots. Of these lots, 27% also yielded seedlings expressing bacterial fruit blotch symptoms when planted under conditions of 30 degrees C and 90% relative humidity. A. avenae subsp. citrulli was detected and recovered from the pulp of 33 and 19%, respectively, of symptomless fruit whose blossoms were inoculated with A. avenae subsp. citrulli. The ability to penetrate watermelon flowers was not unique to A. avenae subsp. citrulli, because blossoms inoculated with Pantoea ananatis also resulted in infested seed and pulp. The data indicate that watermelon blossoms are a potential site of ingress for fruit and seed infestation by A. avenae subsp. citrulli.

Impact of endoscopic stent insertion on detection of viable circulating tumor cells from obstructive colorectal cancer.

Science.gov (United States)

Yamashita, Shinya; Tanemura, Masahiro; Sawada, Genta; Moon, Jeongho; Shimizu, Yosuke; Yamaguchi, Toshiki; Kuwai, Toshio; Urata, Yasuo; Kuraoka, Kazuya; Hatanaka, Nobutaka; Yamashita, Yoshinori; Taniyama, Kiyomi

2018-01-01

The placement of a self-expanding metallic stent (SEMS) in obstructive colorectal cancer (OCRC) is acknowledged to be a safe and effective procedure for the relief of obstruction. However, there is concern that shear forces acting on the tumor during stent expansion may release cancer cells into the circulation, resulting in a poor prognosis. The aim of the present study was to determine whether colonic stent insertion increases viable circulating tumor cells (v-CTCs). A telomerase-specific replication-selective adenovirus-expressing GFP (TelomeScanF35) detection system was used to detect v-CTCs in 8 OCRC patients with a SEMS before and after stent insertion and after surgical resection. In 7 patients, a SEMS was inserted as a bridge to surgery (BTS), and in one patient, a SEMS was inserted for palliation. Surgical resection (R0) was performed in 7 patients. Four patients had no v-CTCs before SEMS placement, two of four measurable patients had an increased number of v-CTCs after SEMS placement (1-3 v-CTCs), and one of two patients with increased v-CTCs developed distant lymphatic metastasis despite curative resection. Four patients had v-CTCs (1-19 cells) before SEMS placement, and two of these four patients had an increase in the number of v-CTCs (20-21 cells) after SEMS placement, while one of the four patients died early with distant metastasis. The present study demonstrated that endoscopic stent insertion for OCRC may result in tumor cell dissemination into the peripheral circulation and may induce distant metastases.

Sun drying of residual annatto seed powder

Directory of Open Access Journals (Sweden)

Dyego da Costa Santos

2015-01-01

Full Text Available Residual annatto seeds are waste from bixin extraction in the food, pharmaceutical and cosmetic industries. Most of this by-product is currently discarded; however, the use of these seeds in human foods through the elaboration of powder added to other commercial powders is seen as a viable option. This study aimed at drying of residual annatto powder, with and without the oil layer derived from the industrial extraction of bixin, fitting different mathematical models to experimental data and calculating the effective moisture diffusivity of the samples. Powder containing oil exhibited the shortest drying time, highest drying rate (≈ 5.0 kg kg-1 min-1 and highest effective diffusivity (6.49 × 10-12 m2 s-1. All mathematical models assessed were a suitable representation of the drying kinetics of powders with and without oil, with R2 above 0.99 and root mean square error values lower than 1.0.

Bioinspired seeding of biomaterials using three dimensional microtissues induces chondrogenic stem cell differentiation and cartilage formation under growth factor free conditions

NARCIS (Netherlands)

Leijten, Jeroen Christianus Hermanus; Moreira Teixeira, Liliana; Bolander, J.; Ji, W.; Vanspauwen, B.; Lammertyn, J.; Schrooten, J.; Luyten, F.P.

2016-01-01

Cell laden biomaterials are archetypically seeded with individual cells and steered into the desired behavior using exogenous stimuli to control growth and differentiation. In contrast, direct cell-cell contact is instructive and even essential for natural tissue formation. Namely, microaggregation

Repair of Traumatic Skeletal Muscle Injury with Bone-Marrow-Derived Mesenchymal Stem Cells Seeded on Extracellular Matrix

Science.gov (United States)

2010-06-02

expressing full length dystrophin can complement Duchenne muscular dystrophy myotubes by cell fusion. Hum Mol Genet 15, 213, 2006. 52. Pittenger, M.F., et al... muscle , and vascular tissue, that are necessary for viable muscular regeneration after muscle defect injury.29–32 Cells from the bone marrow are known to...3,3-diaminobenzidine. Muscular infiltration into the ECM was further confirmed by immunofluorescent staining for the muscle -specific cyto- skeleton

Dynamic Subcellular Localization of Iron during Embryo Development in Brassicaceae Seeds

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Miguel A. Ibeas

2017-12-01

Full Text Available Iron is an essential micronutrient for plants. Little is know about how iron is loaded in embryo during seed development. In this article we used Perls/DAB staining in order to reveal iron localization at the cellular and subcellular levels in different Brassicaceae seed species. In dry seeds of Brassica napus, Nasturtium officinale, Lepidium sativum, Camelina sativa, and Brassica oleracea iron localizes in vacuoles of cells surrounding provasculature in cotyledons and hypocotyl. Using B. napus and N. officinale as model plants we determined where iron localizes during seed development. Our results indicate that iron is not detectable by Perls/DAB staining in heart stage embryo cells. Interestingly, at torpedo development stage iron localizes in nuclei of different cells type, including integument, free cell endosperm and almost all embryo cells. Later, iron is detected in cytoplasmic structures in different embryo cell types. Our results indicate that iron accumulates in nuclei in specific stages of embryo maturation before to be localized in vacuoles of cells surrounding provasculature in mature seeds.

Exploring the Anticancer Activity of Grape Seed Extract on Skin Cancer Cell Lines A431

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V. Mohansrinivasan

2015-08-01

Full Text Available In this study, grape seeds were extracted using ethyl acetate and petroleum ether by solvent-solvent extraction method. The phytochemical tests were performed to identify different phytochemical compounds present in the grape seed extract (GSE. Antibacterial activity of the GSE was determined using agar diffusion method against Gram- positive and Gram-negative bacteria. Gas chromatography-mass spectrometry (GC-MS and Fourier transform infrared spectroscopy (FTIR analysis was done to identify the presence of bioactive compounds and their functional groups. The GC-MS results revealed a total of four compounds, known to have potent activity against cancer cells, viz, squalene, the most potent compound found in ethyl acetate extract and diethyl phthalate, ethyl-9- cis -11- trans octadecadienoate and (R-(--14,-methyl-8-Hexadecyn-1-ol in petroleum ether extract. Cytotoxic activity of the GSE was observed against skin cancer cell lines A4321 using 3-(4, 5-dimethylthiazol-2-yl-2-5-diphenyl tetrazolium bromide MTT assay. The IC50 value of the GSE against A431 skin cancer cell line was 480 µg/mL. This is first such report against A4321 cell lines. The study gives the overall perception about importance of GSE in medicine and nutraceuticals purposes.

Sticking to cellulose: exploiting Arabidopsis seed coat mucilage to understand cellulose biosynthesis and cell wall polysaccharide interactions.

Science.gov (United States)

Griffiths, Jonathan S; North, Helen M

2017-05-01

The cell wall defines the shape of cells and ultimately plant architecture. It provides mechanical resistance to osmotic pressure while still being malleable and allowing cells to grow and divide. These properties are determined by the different components of the wall and the interactions between them. The major components of the cell wall are the polysaccharides cellulose, hemicellulose and pectin. Cellulose biosynthesis has been extensively studied in Arabidopsis hypocotyls, and more recently in the mucilage-producing epidermal cells of the seed coat. The latter has emerged as an excellent system to study cellulose biosynthesis and the interactions between cellulose and other cell wall polymers. Here we review some of the major advances in our understanding of cellulose biosynthesis in the seed coat, and how mucilage has aided our understanding of the interactions between cellulose and other cell wall components required for wall cohesion. Recently, 10 genes involved in cellulose or hemicellulose biosynthesis in mucilage have been identified. These discoveries have helped to demonstrate that xylan side-chains on rhamnogalacturonan I act to link this pectin directly to cellulose. We also examine other factors that, either directly or indirectly, influence cellulose organization or crystallization in mucilage. © 2017 INRA. New Phytologist © 2017 New Phytologist Trust.

Enhanced photovoltaic performance of ZnO nanorod-based dye-sensitized solar cells by using Ga doped ZnO seed layer

Energy Technology Data Exchange (ETDEWEB)

Dou, Yuanyao [State Key Laboratory of Mechanical Transmission, Chongqing University, Chongqing 400044 (China); Department of Applied Physics, College of Physics, Chongqing University, Chongqing 401331 (China); Wu, Fang, E-mail: fang01234@163.com [State Key Laboratory of Mechanical Transmission, Chongqing University, Chongqing 400044 (China); Department of Applied Physics, College of Physics, Chongqing University, Chongqing 401331 (China); Mao, Caiying [Department of Applied Physics, College of Physics, Chongqing University, Chongqing 401331 (China); Fang, Liang, E-mail: lfang@cqu.edu.cn [State Key Laboratory of Mechanical Transmission, Chongqing University, Chongqing 400044 (China); Department of Applied Physics, College of Physics, Chongqing University, Chongqing 401331 (China); Guo, Shengchun [Department of Applied Physics, College of Physics, Chongqing University, Chongqing 401331 (China); Zhou, Miao [State Key Laboratory of Mechanical Transmission, Chongqing University, Chongqing 400044 (China)

2015-06-05

Highlights: • ZnO nanorods were grown on Ga-doped ZnO seed layers using hydrothermal method. • Using the ZnO nanorods as photoanodes for fabricated dye-sensitized solar cells. • The highest η of 1.23% can be achieved in a DSSC with 3 at.% Ga-doped in seeds. • The effects of ZnO seed layers on electron transport properties were investigated. • The enhancement performance of DSSCs contributed to higher dye loading and η{sub cc}. - Abstract: Zinc oxide (ZnO) nanorod arrays were grown on FTO substrates with a Ga-doped ZnO (GZO) seed layer by a hydrothermal method. GZO seed layers were obtained via sol–gel technology with Ga concentration in the range of 0–4 at.%. The dye sensitized solar cells (DSSCs) using ZnO nanorod arrays as the photoanode layers were prepared. The effect of Ga dopant concentrations in ZnO seed layer on the morphology features of ZnO nanorod arrays and the performance of DSSCs were systematically investigated. Results indicate that the average diameter and density of ZnO nanorod arrays decrease with increasing Ga concentration, but their length shows an opposite trend. The photocurrent density–voltage (J–V) characteristics reveal that the DSSCs with GZO seed layer exhibit significantly improved photovoltaic performance. In particular, the highest energy conversion efficiency (η) of 1.23% can be achieved in a DSSC with 3 at.% Ga doping, which is increased by 86.36% compared with that of the undoped DSSC. The external quantum efficiency (EQE) spectra and electrochemical impedance spectroscopy (EIS) were employed to explore the photon-to-electron conversion process in DSSCs. It is demonstrated that the performance enhancement of DSSCs based on GZO seed layer can be attributed to higher amount of dye loading, more efficient electron transportation and better electrons collection efficiency.

Molecular dynamics in germinating, endophyte-colonized quinoa seeds

Science.gov (United States)

2017-01-01

Aims The pseudo-cereal quinoa has an outstanding nutritional value. Seed germination is unusually fast, and plant tolerance to salt stress exceptionally high. Seemingly all seeds harbor bacterial endophytes. This work examines mitogen-activated protein kinase (MAPK) activities during early development. It evaluates possible contribution of endophytes to rapid germination and plant robustness. Methods MAPK activities were monitored in water- and NaCl-imbibed seeds over a 4-h-period using an immunoblot-based approach. Cellulolytic and pectinolytic abilities of bacteria were assessed biochemically, and cellular movement, biofilm, elicitor and antimicrobial compound synthesis genes sequenced. GyrA-based, cultivation-independent studies provided first insight into endophyte diversity. Results Quinoa seeds and seedlings exhibit remarkably complex and dynamic MAPK activity profiles. Depending on seed origin, variances exist in MAPK patterns and probably also in endophyte assemblages. Mucilage-degrading activities enable endophytes to colonize seed surfaces of a non-host species, chia, without apparent adverse effects. Conclusions Owing to their motility, cell wall-loosening and elicitor-generating abilities, quinoa endophytes have the potential to drive cell expansion, move across cell walls, generate damage-associated molecular patterns and activate MAPKs in their host. Bacteria may thus facilitate rapid germination and confer a primed state directly upon seed rehydration. Transfer into non-native crops appears both desirable and feasible. PMID:29416180

Evolution of plant cell wall: Arabinogalactan-proteins from three moss genera show structural differences compared to seed plants.

Science.gov (United States)

Bartels, Desirée; Baumann, Alexander; Maeder, Malte; Geske, Thomas; Heise, Esther Marie; von Schwartzenberg, Klaus; Classen, Birgit

2017-05-01

Arabinogalactan-proteins (AGPs) are important proteoglycans of plant cell walls. They seem to be present in most, if not all seed plants, but their occurrence and structure in bryophytes is widely unknown and actually the focus of AGP research. With regard to evolution of plant cell wall, we isolated AGPs from the three mosses Sphagnum sp., Physcomitrella patens and Polytrichastrum formosum. The moss AGPs show structural characteristics common for AGPs of seed plants, but also unique features, especially 3-O-methyl-rhamnose (trivial name acofriose) as terminal monosaccharide not found in arabinogalactan-proteins of angiosperms and 1,2,3-linked galactose as branching point never found in arabinogalactan-proteins before. Copyright © 2017 Elsevier Ltd. All rights reserved.

Induction of micronuclei in the root tip cells of Haplopappus germinating seeds by fission neutrons and X rays

International Nuclear Information System (INIS)

Hanmoto, Hidehiro; Yonezawa, Yoshihiko; Itoh, Tetsuo; Kondo, Sohei.

1992-01-01

Seeds of Haplopappus gracilis (2n=4), an annual Compositae, were soaked in water for 24 hr and then irradiated with fission neutrons from the 1-wattage reactor, UTR-KINKI, or X rays. The root tip cells were inspected at 48 hr post-irradiation for evidence of chromosome damage using micronucleus as endpoint. The frequency of neutron-induced micronuclei increased almost linearly as the dose increased up to as much as 1.2 Gy. X-ray-induced micronuclei showed an exponential dose-response relation. From dose-response data, we estimated that the dose necessary to induce micronuclei at a frequency of 5 per 1,000 cells was 1.2 Gy for neutrons and 8.6 Gy for X rays. Thus, to induce chromosome damage in the somatic cells of germinating Haplopappus seeds, fission neutrons were much more effective than X rays. (author)

Using composting for control seed germination of invasive plant (water hyacinth) in Extremadura (Spain)

Science.gov (United States)

Labrador, Juana; Gordillo, Judit; Ruiz, Trinidad; Albano, Eva; Moreno, Marta M.

2016-04-01

The biotransformation of the invasive water hyacinth (Eichhornia crassipes) by composting has been showed as a viable alternative to offset the economic cost of eliminating an invasive plant giving a value to the by-product; however, as result of the propagative plant capacity, it was necessary to check if the composting process could eliminate the germination seed rate. Despite the high temperatures and the biochemical biotransformation processes of the composting components, in the case of seed water hyacinth, with a recovery rate of 100%, damage was observed in some parts of the seed anatomy such as in the outer teguments; however, other parts of the seed coat and the endosperm maintained their integrity. A microscopic analysis revealed that the embryo was noticeable and this was supported by the rate of seed germination observed (3.5 ± 0.96%). The results indicate that the use of water hyacinth for compost production is not completely safe from an environmental perspective. Keywords: Eichhornia crassipes, water hyacinth, invasive plant, seed anatomy, seed germination rate, compost. References: Ruiz T., Martín de Rodrigo E., Lorenzo G., Albano E., Morán R., Sánchez J.M. 2008. The Water Hyacinth, Eichhornia crassipes: an invasive plant in the Guadiana River Basin (Spain). Aquatic Invasions Volume 3, Issue 1:42-53.

A propidium monoazide–quantitative PCR method for the detection and quantification of viable Enterococcus faecalis in large-volume samples of marine waters

KAUST Repository

Salam, Khaled W.; El-Fadel, Mutasem E.; Barbour, Elie K.; Saikaly, Pascal

2014-01-01

The development of rapid detection assays of cell viability is essential for monitoring the microbiological quality of water systems. Coupling propidium monoazide with quantitative PCR (PMA-qPCR) has been successfully applied in different studies for the detection and quantification of viable cells in small-volume samples (0.25-1.00 mL), but it has not been evaluated sufficiently in marine environments or in large-volume samples. In this study, we successfully integrated blue light-emitting diodes for photoactivating PMA and membrane filtration into the PMA-qPCR assay for the rapid detection and quantification of viable Enterococcus faecalis cells in 10-mL samples of marine waters. The assay was optimized in phosphate-buffered saline and seawater, reducing the qPCR signal of heat-killed E. faecalis cells by 4 log10 and 3 log10 units, respectively. Results suggest that high total dissolved solid concentration (32 g/L) in seawater can reduce PMA activity. Optimal PMA-qPCR standard curves with a 6-log dynamic range and detection limit of 102 cells/mL were generated for quantifying viable E. faecalis cells in marine waters. The developed assay was compared with the standard membrane filter (MF) method by quantifying viable E. faecalis cells in seawater samples exposed to solar radiation. The results of the developed PMA-qPCR assay did not match that of the standard MF method. This difference in the results reflects the different physiological states of E. faecalis cells in seawater. In conclusion, the developed assay is a rapid (∼5 h) method for the quantification of viable E. faecalis cells in marine recreational waters, which should be further improved and tested in different seawater settings. © 2014 Springer-Verlag Berlin Heidelberg.

A propidium monoazide–quantitative PCR method for the detection and quantification of viable Enterococcus faecalis in large-volume samples of marine waters

KAUST Repository

Salam, Khaled W.

2014-08-23

The development of rapid detection assays of cell viability is essential for monitoring the microbiological quality of water systems. Coupling propidium monoazide with quantitative PCR (PMA-qPCR) has been successfully applied in different studies for the detection and quantification of viable cells in small-volume samples (0.25-1.00 mL), but it has not been evaluated sufficiently in marine environments or in large-volume samples. In this study, we successfully integrated blue light-emitting diodes for photoactivating PMA and membrane filtration into the PMA-qPCR assay for the rapid detection and quantification of viable Enterococcus faecalis cells in 10-mL samples of marine waters. The assay was optimized in phosphate-buffered saline and seawater, reducing the qPCR signal of heat-killed E. faecalis cells by 4 log10 and 3 log10 units, respectively. Results suggest that high total dissolved solid concentration (32 g/L) in seawater can reduce PMA activity. Optimal PMA-qPCR standard curves with a 6-log dynamic range and detection limit of 102 cells/mL were generated for quantifying viable E. faecalis cells in marine waters. The developed assay was compared with the standard membrane filter (MF) method by quantifying viable E. faecalis cells in seawater samples exposed to solar radiation. The results of the developed PMA-qPCR assay did not match that of the standard MF method. This difference in the results reflects the different physiological states of E. faecalis cells in seawater. In conclusion, the developed assay is a rapid (∼5 h) method for the quantification of viable E. faecalis cells in marine recreational waters, which should be further improved and tested in different seawater settings. © 2014 Springer-Verlag Berlin Heidelberg.

Galactomannan from Schizolobium amazonicum seed and its sulfated derivatives impair metabolism in HepG2 cells.

Science.gov (United States)

Cunha de Padua, Monique Meyenberg; Suter Correia Cadena, Silvia Maria; de Oliveira Petkowicz, Carmen Lucia; Martinez, Glaucia Regina; Rodrigues Noleto, Guilhermina

2017-08-01

This study evaluated the effects of native galactomannan from Schizolobium amazonicum seeds and its sulfated forms on certain metabolic parameters of HepG2 cells. Aqueous extraction from S. amazonicum seeds furnished galactomannan with 3.2:1 Man:Gal ratio (SAGM) and molar mass of 4.34×10 5 g/mol. The SAGM fraction was subjected to sulfation using chlorosulfonic acid to obtain SAGMS1 and SAGMS2 with DS of 0.4 and 0.6, respectively. Cytotoxicity of SAGM, SAGMS1, and SAGMS2 was evaluated in human hepatocellular carcinoma cells (HepG2). After 72h, SAGM decreased the viability of HepG2 cells by 50% at 250μg/mL, while SAGMS1 reduced it by 30% at the same concentration. SAGM, SAGMS1, and SAGMS2 promoted a reduction in oxygen consumption and an increase in lactate production in non-permeabilized HepG2 cells after 72h of treatment. These results suggest that SAGM, SAGMS1, and SAGMS2 could be recognized by HepG2 cells and might trigger alterations that impair its survival. These effects could be implicated in the modification of the oxidative phosphorylation process in HepG2 cells and activation of the glycolytic pathway. Copyright © 2017 Elsevier B.V. All rights reserved.

Electron beam irradiation: laboratory and field studies of cowpea seeds

International Nuclear Information System (INIS)

Srinivasan, K.; Chauhan, S.K.; Prasad, T.V.; Pramod, R.; Verma, V.P.; Petwal, V.; Dwivedi, J.; Bhalla, S.

2015-01-01

Cowpea (Vigna unguiculata) rich in protein and vitamins is emerging as one of the most important food legumes to tackle malnutrition. Pulse beetles (Callosobruchus chinensis and C. maculatus) are the pests of economic importance causing enormous losses during storage. Although various pest management strategies exist for the control of these pests, environmental concerns necessitate developing ecofriendly strategies. Electron beam (EB) irradiation has the potential to be a viable, non-chemical, residue-free strategy for management of pulse beetles during storage, but higher doses affect seed germination and viability. Hence, the present investigation was taken up to analyse the dosage effect of the irradiation on seed attributes of cowpea. Healthy cowpea seeds were irradiated with low energy electrons at different doses viz., 180, 360, 540, 720, 900, 1080, 1260, 1440 and 1620 Gy at 500 keV using the EB Accelerator facility at Raja Ramanna Centre for Advanced Technology, Indore. EB irradiated seeds were tested for physiological viz., germination, seedling vigour and vigour index and biochemical parameters viz., electrical conductivity of seed leachate, seed viability/tetrazolium test and dehydrogenase activity. Germination and vigour of the irradiated seeds were evaluated as per the ISTA Rules (ISTA, 1996). Vigour index was calculated as the product of germination percentage and seedling vigour. About 3,000 irradiated seeds from each dose were grown in the field at the Experimental farm, National Bureau of Plant Genetic Resources, New Delhi. Seeds harvested from 1500 individual plants of M 1 generation from each dose (50 seeds from each plant individually) were sown in next season and observed for chlorophyll mutations, if any. Results revealed that doses upto 1080 Gy (88%) did not affect the germination of cowpea seeds drastically as compared to untreated seeds (98%). Lower doses viz., 180 and 360 Gy had no impact on vigour components while higher doses (1080 Gy

Endoreduplication intensity as a marker of seed developmental stage in the Fabaceae.

Science.gov (United States)

Rewers, Monika; Sliwinska, Elwira

2012-12-01

Flow cytometry (FCM) can be used to study cell cycle activity in developing, mature and germinating seeds. It provides information about a seed's physiological state and therefore can be used by seed growers for assessing optimal harvest times and presowing treatments. Because an augmented proportion of 4C nuclei usually is indicative of high mitotic activity, the 4C/2C ratio is commonly used to follow the progress of seed development and germination. However, its usefulness for polysomatic (i.e., containing cells with different DNA content) seeds is questioned. Changes in cell cycle/endoreduplication activity in developing seeds of five members of the Fabaceae were studied to determine a more suitable marker of seed developmental stages for polysomatic species based on FCM measurements. Seeds of Phaseolus vulgaris, Medicago sativa, Pisum sativum, Vicia sativa, and Vicia faba var. minor were collected 20, 30, 40, 50, and 60 days after flowering (DAF), embryos were isolated and the proportion of nuclei with different DNA contents in the embryo axis and cotyledon was established. The ratios 4C/2C and (Σ>2C)/2C were calculated. Dried seeds were subjected to laboratory germination tests following international seed testing association (ISTA) rules. Additionally, the absolute nuclear DNA content was estimated in the leaves of the studied species. During seed development nuclei with DNA contents from 2C to 128C were detected; the endopolyploidy pattern depended on the species, seed organ and developmental stage. The cell cycle/endoreduplication parameters correlated negatively with genome size. The (Σ>2C)/2C ratio in the cotyledons reflected the seed developmental stage and corresponded with seed germinability. Therefore, this ratio is recommended as a marker in polysomatic seed research and production instead of the 4C/2C ratio, which does not consider the occurrence of endopolyploid cells. Copyright © 2012 International Society for Advancement of Cytometry.

HIGHLY METHYL ESTERIFIED SEEDS is a pectin methyl esterase involved in embryo development.

Science.gov (United States)

Levesque-Tremblay, Gabriel; Müller, Kerstin; Mansfield, Shawn D; Haughn, George W

2015-03-01

Homogalacturonan pectin domains are synthesized in a highly methyl-esterified form that later can be differentially demethyl esterified by pectin methyl esterase (PME) to strengthen or loosen plant cell walls that contain pectin, including seed coat mucilage, a specialized secondary cell wall of seed coat epidermal cells. As a means to identify the active PMEs in seed coat mucilage, we identified seven PMEs expressed during seed coat development. One of these, HIGHLY METHYL ESTERIFIED SEEDS (HMS), is abundant during mucilage secretion, peaking at 7 d postanthesis in both the seed coat and the embryo. We have determined that this gene is required for normal levels of PME activity and homogalacturonan methyl esterification in the seed. The hms-1 mutant displays altered embryo morphology and mucilage extrusion, both of which are a consequence of defects in embryo development. A significant decrease in the size of cells in the embryo suggests that the changes in embryo morphology are a consequence of lack of cell expansion. Progeny from a cross between hms-1 and the previously characterized PME inhibitor5 overexpression line suggest that HMS acts independently from other cell wall-modifying enzymes in the embryo. We propose that HMS is required for cell wall loosening in the embryo to facilitate cell expansion during the accumulation of storage reserves and that its role in the seed coat is masked by redundancy. © 2015 American Society of Plant Biologists. All Rights Reserved.

Seed quality in informal seed systems

NARCIS (Netherlands)

Biemond, P.C.

2013-01-01

Keywords: informal seed systems, seed recycling, seed quality, germination, seed pathology, seed health, seed-borne diseases, mycotoxigenic fungi, Fusarium verticillioides, mycotoxins, Vigna unguiculata, Zea mays, Nigeria.

Seed is a crucial input for agricultural production.

Proximate composition, extraction, characterization and comparative assessment of coconut (Cocos nucifera) and melon (Colocynthis citrullus) seeds and seed oils.

Science.gov (United States)

Obasi, N A; Ukadilonu, Joy; Eze, Eberechukwu; Akubugwo, E I; Okorie, U C

2012-01-01

Proximate composition, extraction, characterization and comparative assessment of Cocos nucifera and Colocynthis citrullus seeds and seed oils were evaluated in this work using standard analytical techniques. The results showed the percentage (%) moisture, crude fibre, ash, crude protein, lipids and total carbohydrate contents of the seeds as 7.51 and 4.27, 7.70 and 5.51, 1.02 and 2.94, 10.57 and 11.67, 47.80 and 50.42 and 32.84 and 29.47 while the calorific values were 553.99 and 567.32 Kcal/100 g for C. nucifera and C. citrullus, respectively. The two seed oils were odourless and at room temperature (30 degrees C) liquids, with a pale yellow to yellowish colouration. Lipid indices of the seed oils indicated the Acid Values (AV) as 2.06-6.36 mg NaOH g(-1) and 2.99-6.17 mg NaOH g(-1), Free Fatty Acids (FFA) as 1.03-3.18 and 1.49-3.09%, Saponification Values (SV) as 252.44-257.59 and 196.82-201.03 mg KOH g(-1), Iodine Values (IV) as 9.73-10.99 and 110.93-111.46 mg of I2 g(-1) of oil and Peroxide Values (PV) as 0.21-0.21 and 1.53-2.72 mg O2 kg(-1) for soxhlet-mechanical extracted C. nucifera and C. citrullus seed oils, respectively. The studied characteristics of the oil extracts in most cases compared favourably with most conventional vegetable oils sold in the Nigeria markets; however, there were some observed levels of significant differences in the values at p < or = 0.05. These results suggest that the seeds examined may be nutritionally potent and also viable sources of seed oils judging by their oil yield. The data also showed that the seed oils were edible inferring from their low AV and their corresponding low FFA contents. Industrially, the results revealed the seed oils to have great potentials in soap manufacturing industries because of their high SV. They were also shown to be non-drying due to their low IV which also suggested that the oils contain few unsaturated bonds and therefore have low susceptibility to oxidative rancidity and deterioration as

Bio-electrosprayed multicellular zebrafish embryos are viable and develop normally

International Nuclear Information System (INIS)

Clarke, Jonathan D W; Jayasinghe, Suwan N

2008-01-01

Bio-electrosprays are rapidly emerging as a viable protocol for directly engineering living cells. This communication reports the bio-electrospraying of multicellular organisms, namely zebrafish embryos. The results demonstrate that the bio-electrospray protocol fails to induce any embryological perturbations. In addition to analysing overall embryo morphology, we use transgenic embryos that express green fluorescent protein in specific brain neurons to determine that neuronal numbers and organization are completely normal. These results demonstrate that the bio-electrospraying protocol does not interfere with the complex gene regulation and cell movements required for the development of a multicellular organism. (communication)

Up-regulating the abscisic acid inactivation gene ZmABA8ox1b contributes to seed germination heterosis by promoting cell expansion.

Science.gov (United States)

Li, Yangyang; Wang, Cheng; Liu, Xinye; Song, Jian; Li, Hongjian; Sui, Zhipeng; Zhang, Ming; Fang, Shuang; Chu, Jinfang; Xin, Mingming; Xie, Chaojie; Zhang, Yirong; Sun, Qixin; Ni, Zhongfu

2016-04-01

Heterosis has been widely used in agriculture, but the underlying molecular principles are still largely unknown. During seed germination, we observed that maize (Zea mays) hybrid B73/Mo17 was less sensitive than its parental inbred lines to exogenous abscisic acid (ABA), and endogenous ABA content in hybrid embryos decreased more rapidly than in the parental inbred lines. ZmABA8ox1b, an ABA inactivation gene, was consistently more highly up-regulated in hybrid B73/Mo17 than in its parental inbred lines at early stages of seed germination. Moreover, ectopic expression of ZmABA8ox1b obviously promoted seed germination in Arabidopsis Remarkably, microscopic observation revealed that cell expansion played a major role in the ABA-mediated maize seed germination heterosis, which could be attributed to the altered expression of cell wall-related genes. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Green synthesis of silver nanoparticles using Pimpinella anisum seeds: antimicrobial activity and cytotoxicity on human neonatal skin stromal cells and colon cancer cells

Directory of Open Access Journals (Sweden)

AlSalhi MS

2016-09-01

Full Text Available Mohamad S AlSalhi,1,2 Sandhanasamy Devanesan,1,2 Akram A Alfuraydi,3 Radhakrishnan Vishnubalaji,4 Murugan A Munusamy,3 Kadarkarai Murugan,5 Marcello Nicoletti,6 Giovanni Benelli7 1Research Chair in Laser Diagnosis of Cancers, 2Department of Physics and Astronomy, 3Department of Botany and Microbiology, College of Science, 4Stem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh, Kingdom of Saudi Arabia; 5Division of Entomology, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore, India; 6Department of Environmental Biology, Sapienza University of Rome, Rome, 7Department of Agriculture, Food and Environment, University of Pisa, Pisa, Italy Background: The present study focused on a simple and eco-friendly method for the synthesis of silver nanoparticles (AgNPs with multipurpose anticancer and antimicrobial activities. Materials and methods: We studied a green synthesis route to produce AgNPs by using an aqueous extract of Pimpinella anisum seeds (3 mM. Their antimicrobial activity and cytotoxicity on human neonatal skin stromal cells (hSSCs and colon cancer cells (HT115 were assessed. Results: A biophysical characterization of the synthesized AgNPs was realized: the morphology of AgNPs was determined by transmission electron microscopy, energy dispersive spectroscopy, X-ray powder diffraction, and ultraviolet-vis absorption spectroscopy. Transmission electron microscopy showed spherical shapes of AgNPs of P. anisum seed extracts with a 3.2 nm minimum diameter and average diameter ranging from 3.2 to 16 nm. X-ray powder diffraction highlighted the crystalline nature of the nanoparticles, ultraviolet-vis absorption spectroscopy was used to monitor their synthesis, and Fourier transform infrared spectroscopy showed the main reducing groups from the seed extract. Energy dispersive spectroscopy was used to confirm the presence of elemental silver. We evaluated the antimicrobial potential

Anticancer activity of Nigella sativa (black seed) and its relationship with the thermal processing and quinone composition of the seed.

Science.gov (United States)

Agbaria, Riad; Gabarin, Adi; Dahan, Arik; Ben-Shabat, Shimon

2015-01-01

The traditional preparation process of Nigella sativa (NS) oil starts with roasting of the seeds, an allegedly unnecessary step that was never skipped. The aims of this study were to investigate the role and boundaries of thermal processing of NS seeds in the preparation of therapeutic extracts and to elucidate the underlying mechanism. NS extracts obtained by various seed thermal processing methods were investigated in vitro for their antiproliferative activity in mouse colon carcinoma (MC38) cells and for their thymoquinone content. The effect of the different methods of thermal processing on the ability of the obtained NS oil to inhibit the nuclear factor kappa B (NF-κB) pathway was then investigated in Hodgkin's lymphoma (L428) cells. The different thermal processing protocols yielded three distinct patterns: heating the NS seeds to 50°C, 100°C, or 150°C produced oil with a strong ability to inhibit tumor cell growth; no heating or heating to 25°C had a mild antiproliferative effect; and heating to 200°C or 250°C had no effect. Similar patterns were obtained for the thymoquinone content of the corresponding oils, which showed an excellent correlation with the antiproliferative data. It is proposed that there is an oxidative transition mechanism between quinones after controlled thermal processing of the seeds. While NS oil from heated seeds delayed the expression of NF-κB transcription, non-heated seeds resulted in only 50% inhibition. The data indicate that controlled thermal processing of NS seeds (at 50°C-150°C) produces significantly higher anticancer activity associated with a higher thymoquinone oil content, and inhibits the NF-κB signaling pathway.

Phytoextract of Indian mustard seeds acts by suppressing the generation of ROS against acetaminophen-induced hepatotoxicity in HepG2 cells.

Science.gov (United States)

Parikh, Harita; Pandita, Nancy; Khanna, Aparna

2015-07-01

Indian mustard [Brassica juncea (L.) Czern. & Coss. (Brassicaceae)] is reported to possess diverse pharmacological properties. However, limited information is available concerning its hepatoprotective activity and mechanism of action. To study the protective mechanism of mustard seed extract against acetaminophen (APAP) toxicity in a hepatocellular carcinoma (HepG2) cell line. Hepatotoxicity models were established using APAP (2.5-22.5 mM) based on the cytotoxicity profile. An antioxidant-rich fraction from mustard seeds was extracted and evaluated for its hepatoprotective potential. The mechanism of action was elucidated using various in vitro antioxidant assays, the detection of intracellular generation of reactive oxygen species (ROS), and cell cycle analysis. The phytoconstituents isolated via HPLC-DAD were also evaluated for hepatoprotective activity. Hydromethanolic seed extract exhibited hepatoprotective activity in post- and pre-treatment models of 20 mM APAP toxicity and restored the elevated levels of liver indices to normal values (p DAD analysis revealed the presence quercetin, vitamin E, and catechin, which exhibited hepatoprotective activity. A phytoextract of mustard seeds acts by suppressing the generation of ROS in response to APAP toxicity.

A factor converting viable but nonculturable Vibrio cholerae to a culturable state in eukaryotic cells is a human catalase.

Science.gov (United States)

Senoh, Mitsutoshi; Hamabata, Takashi; Takeda, Yoshifumi

2015-08-01

In our previous work, we demonstrated that viable but nonculturable (VBNC) Vibrio cholerae O1 and O139 were converted to culturable by coculture with eukaryotic cells. Furthermore, we isolated a factor converting VBNC V. cholerae to culturable (FCVC) from a eukaryotic cell line, HT-29. In this study, we purified FCVC by successive column chromatographies comprising UNO Q-6 anion exchange, Bio-Scale CHT2-1 hydroxyapatite, and Superdex 200 10/300 GL. Homogeneity of the purified FCVC was demonstrated by SDS-PAGE. Nano-LC MS/MS analysis showed that the purified FCVC was a human catalase. An experiment of RNAi knockdown of catalase mRNA from HT-29 cells and treatment of the purified FCVC with a catalase inhibitor, 3-amino-1,2,4-triazole confirmed that the FCVC was a catalase. A possible role of the catalase in converting a VBNC V. cholerae to a culturable state in the human intestine is discussed. © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Viable Bacteria Associated with Red Blood Cells and Plasma in Freshly Drawn Blood Donations

DEFF Research Database (Denmark)

Damgaard, Christian; Magnussen, Karin; Enevold, Christian

2015-01-01

) or blue lactose plates. For identification colony PCR was performed using primers targeting 16S rDNA. SETTING: Blood donors attending Capital Region Blood Bank, Copenhagen University Hospital, Rigshospitalet, Hvidovre, Denmark, October 29th to December 10th 2013. PARTICIPANTS: 60 donors (≥50 years old....... CONCLUSIONS: Viable bacteria are present in blood from donors self-reported as medically healthy, indicating that conventional test systems employed by blood banks insufficiently detect bacteria in plasma. Further investigation is needed to determine whether routine testing for anaerobic bacteria and testing......OBJECTIVES: Infection remains a leading cause of post-transfusion mortality and morbidity. Bacterial contamination is, however, detected in less than 0.1% of blood units tested. The aim of the study was to identify viable bacteria in standard blood-pack units, with particular focus on bacteria from...

Molecular Mechanisms Underlying Abscisic Acid/Gibberellin Balance in the Control of Seed Dormancy and Germination in Cereals

Directory of Open Access Journals (Sweden)

Pham A. Tuan

2018-05-01

Full Text Available Seed dormancy is an adaptive trait that does not allow the germination of an intact viable seed under favorable environmental conditions. Non-dormant seeds or seeds with low level of dormancy can germinate readily under optimal environmental conditions, and such a trait leads to preharvest sprouting, germination of seeds on the mother plant prior to harvest, which significantly reduces the yield and quality of cereal crops. High level of dormancy, on the other hand, may lead to non-uniform germination and seedling establishment. Therefore, intermediate dormancy is considered to be a desirable trait as it prevents the problems of sprouting and allows uniformity of postharvest germination of seeds. Induction, maintenance, and release of seed dormancy are complex physiological processes that are influenced by a wide range of endogenous and environmental factors. Plant hormones, mainly abscisic acid (ABA and gibberellin (GA, are the major endogenous factors that act antagonistically in the control of seed dormancy and germination; ABA positively regulates the induction and maintenance of dormancy, while GA enhances germination. Significant progress has been made in recent years in the elucidation of molecular mechanisms regulating ABA/GA balance and thereby dormancy and germination in cereal seeds, and this review summarizes the current state of knowledge on the topic.

Molecular Mechanisms Underlying Abscisic Acid/Gibberellin Balance in the Control of Seed Dormancy and Germination in Cereals

Science.gov (United States)

Tuan, Pham A.; Kumar, Rohit; Rehal, Pawanpuneet K.; Toora, Parneet K.; Ayele, Belay T.

2018-01-01

Seed dormancy is an adaptive trait that does not allow the germination of an intact viable seed under favorable environmental conditions. Non-dormant seeds or seeds with low level of dormancy can germinate readily under optimal environmental conditions, and such a trait leads to preharvest sprouting, germination of seeds on the mother plant prior to harvest, which significantly reduces the yield and quality of cereal crops. High level of dormancy, on the other hand, may lead to non-uniform germination and seedling establishment. Therefore, intermediate dormancy is considered to be a desirable trait as it prevents the problems of sprouting and allows uniformity of postharvest germination of seeds. Induction, maintenance, and release of seed dormancy are complex physiological processes that are influenced by a wide range of endogenous and environmental factors. Plant hormones, mainly abscisic acid (ABA) and gibberellin (GA), are the major endogenous factors that act antagonistically in the control of seed dormancy and germination; ABA positively regulates the induction and maintenance of dormancy, while GA enhances germination. Significant progress has been made in recent years in the elucidation of molecular mechanisms regulating ABA/GA balance and thereby dormancy and germination in cereal seeds, and this review summarizes the current state of knowledge on the topic. PMID:29875780

Seed dormancy and germination.

Science.gov (United States)

Penfield, Steven

2017-09-11

Reproduction is a critical time in plant life history. Therefore, genes affecting seed dormancy and germination are among those under strongest selection in natural plant populations. Germination terminates seed dispersal and thus influences the location and timing of plant growth. After seed shedding, germination can be prevented by a property known as seed dormancy. In practise, seeds are rarely either dormant or non-dormant, but seeds whose dormancy-inducing pathways are activated to higher levels will germinate in an ever-narrower range of environments. Thus, measurements of dormancy must always be accompanied by analysis of environmental contexts in which phenotypes or behaviours are described. At its simplest, dormancy can be imposed by the formation of a simple physical barrier around the seed through which gas exchange and the passage of water are prevented. Seeds featuring this so-called 'physical dormancy' often require either scarification or passage through an animal gut (replete with its associated digestive enzymes) to disrupt the barrier and permit germination. In other types of seeds with 'morphological dormancy' the embryo remains under-developed at maturity and a dormant phase exists as the embryo continues its growth post-shedding, eventually breaking through the surrounding tissues. By far, the majority of seeds exhibit 'physiological dormancy' - a quiescence program initiated by either the embryo or the surrounding endosperm tissues. Physiological dormancy uses germination-inhibiting hormones to prevent germination in the absence of the specific environmental triggers that promote germination. During and after germination, early seedling growth is supported by catabolism of stored reserves of protein, oil or starch accumulated during seed maturation. These reserves support cell expansion, chloroplast development and root growth until photoauxotrophic growth can be resumed. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

Health status of alfalfa leafcutting bee larvae (Hymenoptera: Megachilidae) in United States alfalfa seed fields.

Science.gov (United States)

James, R R; Pitts-Singer, T L

2013-12-01

We conducted a broad geographic survey in the northwestern United States to quantify production losses in the alfalfa leafcutting bee (Megachile rotundata (F.), Hymenoptera: Megachilidae), a solitary pollinator used extensively in alfalfa seed production. Viable larvae were found in only 47.1% of the nest cells collected at the end of the season. Most of the rest of the cells contained pollen balls (typified by a provision but no larva; 16.7%), unknown causes of mortality (15.5%), or larvae killed by chalkbrood (8.0%). Prevalence of pollen balls was correlated positively with bee release rates and negatively with alfalfa stand age. The unknown mortality was correlated with the U.S. Department of Agriculture-Plant Hardiness Zone, and thus, some of the mortality may be caused by high temperature extremes, although the nesting season degree-days were not correlated with this mortality. Chalkbrood prevalence was correlated with possible nesting-resource or crowding-related factors, such as the number of bees released per hectare and the number of shelters used, but not with nesting board disinfection practices. Vapona is used to control parasitoids when the parent bees are incubated before release, and use of this fumigant was associated with an increase in both chalkbrood and diapausing offspring, although any reason for these correlations are unknown. This survey quantifies the variation in the quality of alfalfa leafcutting bee cocoons produced across much of the U.S. alfalfa seed production area.

Protein synthesis in the embryo of Pinus thunbergii seed, 2

International Nuclear Information System (INIS)

Yamamoto, Naoaki; Sasaki, Satohiko.

1977-01-01

14 C-Amino acid incorporating activity in the absence of exogenous mRNA was found in a cell-free system from embryos of light-germinated Pinus thunbergii seeds, but not in that from dark-imbibed seed embryos. Template activity in the cell-free system from the light-germinated seed embryos was observed in the ribosome fraction, especially the polyribosome fraction, but not in the 100,000 x g supernatant fraction (s100). These facts suggest that the nature of the block in protein synthesis during the imbibition of seeds in the dark is due to the lack or inactivity of mRNA. The s100 from light-germinated seed embryos was found to be less active in amino acid incorporation than that from dark-imbibed seed embryos. (auth.)

Fuel cells : a viable fossil fuel alternative

Energy Technology Data Exchange (ETDEWEB)

Paduada, M.

2007-02-15

This article presented a program initiated by Natural Resources Canada (NRCan) to develop proof-of-concept of underground mining vehicles powered by fuel cells in order to eliminate emissions. Recent studies on American and Canadian underground mines provided the basis for estimating the operational cost savings of switching from diesel to fuel cells. For the Canadian mines evaluated, the estimated ventilation system operating cost reductions ranged from 29 per cent to 75 per cent. In order to demonstrate the viability of a fuel cell-powered vehicle, NRCan has designed a modified Caterpillar R1300 loader with a 160 kW hybrid power plant in which 3 stacks of fuel cells deliver up to 90 kW continuously, and a nickel-metal hydride battery provides up to 70 kW. The battery subsystem transiently boosts output to meet peak power requirements and also accommodates regenerative braking. Traction for the loader is provided by a brushless permanent magnet traction motor. The hydraulic pump motor is capable of a 55 kW load continuously. The loader's hydraulic and traction systems are operated independently. Future fuel cell-powered vehicles designed by the program may include a locomotive and a utility vehicle. Future mines running their operations with hydrogen-fueled equipment may also gain advantages by employing fuel cells in the operation of handheld equipment such as radios, flashlights, and headlamps. However, the proton exchange membrane (PEM) fuel cells used in the project are prohibitively expensive. The catalytic content of a fuel cell can add hundreds of dollars per kW of electric output. Production of catalytic precious metals will be strongly connected to the scale of use and acceptance of fuel cells in vehicles. In addition, the efficiency of hydrogen production and delivery is significantly lower than the well-to-tank efficiency of many conventional fuels. It was concluded that an adequate hydrogen infrastructure will be required for the mining industry

3,4-Dihydroxybenzaldehyde Derived from Prunus mume Seed Inhibits Oxidative Stress and Enhances Estradiol Secretion in Human Ovarian Granulosa Tumor Cells

International Nuclear Information System (INIS)

Kono, Ryohei; Nomura, Sachiko; Okuno, Yoshiharu; Nakamura, Misa; Maeno, Akihiro; Kagiya, Tomoko; Tokuda, Akihiko; Inada, Ken-ichi; Matsuno, Akira; Utsunomiya, Tomoko; Utsunomiya, Hirotoshi

2014-01-01

Granulosa cells form ovarian follicles and play important roles in the growth and maturation of oocytes. The protection of granulosa cells from cellular injury caused by oxidative stress is an effective therapy for female infertility. We here investigated an effective bioactive compound derived from Prunus mume seed extract that protects granulosa cells from hydrogen peroxide (H 2 O 2 )-induced apoptosis. We detected the bioactive compound, 3,4-dihydroxybenzaldehyde (3,4-DHBA), via bioactivity-guided isolation and found that it inhibited the H 2 O 2 -induced apoptosis of granulosa cells. We also showed that 3,4-DHBA promoted estradiol secretion in granulosa cells and enhanced the mRNA expression levels of steroidogenic factor 1, a promoter of key steroidogenic enzymes. These results suggest that P. mume seed extract may have clinical potential for the prevention and treatment of female infertility

Different modes of hydrogen peroxide action during seed germination

Directory of Open Access Journals (Sweden)

Łukasz eWojtyla

2016-02-01

Full Text Available Hydrogen peroxide was initially recognized as a toxic molecule that causes damage at different levels of cell organization and thus losses in cell viability. From the 1990s, the role of hydrogen peroxide as a signaling molecule in plants has also been discussed. The beneficial role of H2O2 as a central hub integrating signaling network in response to biotic and abiotic stress and during developmental processes is now well established. Seed germination is the most pivotal phase of the plant life cycle, affecting plant growth and productivity. The function of hydrogen peroxide in seed germination and seed aging has been illustrated in numerous studies; however, the exact role of this molecule remains unknown. This review evaluates evidence that shows that H2O2 functions as a signaling molecule in seed physiology in accordance with the known biology and biochemistry of H2O2. The importance of crosstalk between hydrogen peroxide and a number of signaling molecules, including plant phytohormones such as abscisic acid, gibberellins and ethylene and reactive molecules such as nitric oxide and hydrogen sulfide acting on cell communication and signaling during seed germination, is highlighted. The current study also focuses on the detrimental effects of H2O2 on seed biology, i.e., seed aging that leads to a loss of germination efficiency. The dual nature of hydrogen peroxide as a toxic molecule on one hand and as a signal molecule on the other is made possible through the precise spatial and temporal control of its production and degradation. Levels of hydrogen peroxide in germinating seeds and young seedlings can be modulated via pre-sowing seed priming/conditioning. This rather simple method is shown to be a valuable tool for improving seed quality and for enhancing seed stress tolerance during post-priming germination. In this review, we outline how seed priming/conditioning affects the integrative role of hydrogen peroxide in seed germination and

Concerning the role of cell lysis-cryptic growth in anaerobic side-stream reactors: the single-cell analysis of viable, dead and lysed bacteria.

Science.gov (United States)

Foladori, P; Velho, V F; Costa, R H R; Bruni, L; Quaranta, A; Andreottola, G

2015-05-01

In the Anaerobic Side-Stream Reactor (ASSR), part of the return sludge undergoes alternating aerobic and anaerobic conditions with the aim of reducing sludge production. In this paper, viability, enzymatic activity, death and lysis of bacterial cells exposed to aerobic and anaerobic conditions for 16 d were investigated at single-cell level by flow cytometry, with the objective of contributing to the understanding of the mechanisms of sludge reduction in the ASSR systems. Results indicated that total and viable bacteria did not decrease during the anaerobic phase, indicating that anaerobiosis at ambient temperature does not produce a significant cell lysis. Bacteria decay and lysis occurred principally under aerobic conditions. The aerobic decay rate of total bacteria (bTB) was considered as the rate of generation of lysed bacteria. Values of bTB of 0.07-0.11 d(-1) were measured in anaerobic + aerobic sequence. The enzymatic activity was not particularly affected by the transition from anaerobiosis to aerobiosis. Large solubilisation of COD and NH4(+) was observed only under anaerobic conditions, as a consequence of hydrolysis of organic matter, but not due to cell lysis. The observations supported the proposal of two independent mechanisms contributing equally to sludge reduction: (1) under anaerobic conditions: sludge hydrolysis of non-bacterial material, (2) under aerobic conditions: bacterial cell lysis and oxidation of released biodegradable compounds. Copyright © 2015 Elsevier Ltd. All rights reserved.

Managing Viable Knowledge

NARCIS (Netherlands)

Achterbergh, J.M.I.M.; Vriens, D.J.

2002-01-01

In this paper, Beer's Viable System Model (VSM) is applied to knowledge management. Based on the VSM, domains of knowledge are identified that an organization should possess to maintain its viability. The logic of the VSM is also used to support the diagnosis, design and implementation of the

Mutation induction and isolation in potato through true seed and tuber mutagenesis and use of tissue culture

International Nuclear Information System (INIS)

Upadhya, M.D.; Abraham, M.J.; Dass, B.; Chandra, R.

1982-01-01

Advance MV generation clones from hydrazine-sulphate-treated 'O.T' cultures have been field evaluated and 12 cultures have been selected for yield trials. One culture, DN-31-3, has been found to be day-neutral in its tuberization behaviour. Four JL/RA clones have been selected after a large-scale field trial. These clones are the selections from gamma-irradiated self seeds of Kufri Lauvkar (A-7416) and hybrid A-2235. Similarly 15 MV 3 clones have been selected from the populations raised from EMS- and DES-treated self seeds of A-2235. Day-neutral mutants have been selected from the fourth to seventh sprouts taken from EMS-treated tuber halves of Kufri Jyoti. From the sixth sprout harvest from EMS-treated Kufri Jyoti halves, one mutant, BCN-6-2, has been isolated which showed less than 30 cysts of G. rostochiensis in the MV 2 generation. This clone has been multiplied and made disease-free through apical meristem culture. Through the use of a new medium, PM-32, the plating efficiency of mechanically isolated single callus cells of dihaploid PH-258 is 30-35%. Nitsch's medium has been modified to formulate a new medium for direct embryogenesis in single callus cells of Phulwa. All stages up to the globular stage could be observed after five to six weeks of culture. Efforts were continued for the enzymatic isolation of single-leaf mesophyll cells from dihaploid PH-258. A new cell separation medium has been formulated which gives 80% viable cells. The LD 50 and LD 100 doses of EMS for the single callus cells of dihaploid PH-255 were found to be 500 ppm and 1000 ppm respectively. (author)

Seed Anatomy and Water Uptake in Relation to Seed Dormancy in Opuntia tomentosa (Cactaceae, Opuntioideae)

Science.gov (United States)

Orozco-Segovia, A.; Márquez-Guzmán, J.; Sánchez-Coronado, M. E.; Gamboa de Buen, A.; Baskin, J. M.; Baskin, C. C.

2007-01-01

Background and Aims There is considerable confusion in the literature concerning impermeability of seeds with ‘hard’ seed coats, because the ability to take up (imbibe) water has not been tested in most of them. Seeds of Opuntia tomentosa were reported recently to have a water-impermeable seed coat sensu lato (i.e. physical dormancy), in combination with physiological dormancy. However, physical dormancy is not known to occur in Cactaceae. Therefore, the aim of this study was to determine if seeds of O. tomentosa are water-permeable or water-impermeable, i.e. if they have physical dormancy. Methods The micromorphology of the seed coat and associated structures were characterized by SEM and light microscopy. Permeability of the seed-covering layers was assessed by an increase in mass of seeds on a wet substrate and by dye-tracking and uptake of tritiated water by intact versus scarified seeds. Key Results A germination valve and a water channel are formed in the hilum–micropyle region during dehydration and ageing in seeds of O. tomentosa. The funicular envelope undoubtedly plays a role in germination of Opuntia seeds via restriction of water uptake and mechanical resistance to expansion of the embryo. However, seeds do not exhibit any of three features characteristic of those with physical dormancy. Thus, they do not have a water-impermeable layer(s) of palisade cells (macrosclereids) or a water gap sensu stricto and they imbibe water without the seed coat being disrupted. Conclusions Although dormancy in seeds of this species can be broken by scarification, they have physiological dormancy only. Further, based on information in the literature, it is concluded that it is unlikely that any species of Opuntia has physical dormancy. This is the first integrative study of the anatomy, dynamics of water uptake and dormancy in seeds of Cactaceae subfamily Opuntioideae. PMID:17298989

The effectiveness of a standardized rose hip powder, containing seeds and Shells of Rosa canina, on cell longevity, skin wrinkles, moisture, and elasticity 

DEFF Research Database (Denmark)

Phetcharat, L; Wongsuphasawat, K; Winther, Kaj

2015-01-01

Objective: To evaluate the effects of a rose hip powder (Hyben Vital®) made from seeds and shells on cell senescence, skin wrinkling, and aging. Methods: A total of 34 healthy subjects, aged 35–65 years, with wrinkles on the face (crow’s-feet) were subjected to a randomized and double-blinded cli...... hip product on cell membranes of stored erythrocyte cells observed in this study may contribute to improve the cell longevity and obstructing skin aging. Keywords: rose hip seed and shell powder, cell longevity, wrinkles, aging skin, moisture, elasticity......Objective: To evaluate the effects of a rose hip powder (Hyben Vital®) made from seeds and shells on cell senescence, skin wrinkling, and aging. Methods: A total of 34 healthy subjects, aged 35–65 years, with wrinkles on the face (crow’s-feet) were subjected to a randomized and double...... longevity was measured in terms of leakage of hemoglobin through red cell membranes (hemolytic index) in blood samples kept in a blood bank for 5 weeks. Significance of all values was attained with P#0.05. Results: In the double-blinded study, the rose hip group showed statistically significant improvements...

Seed regulations and local seed systems

NARCIS (Netherlands)

Louwaars, N.

2000-01-01

Seed regulations have been introduced in most countries based on the development of formal seed production. Concerns about seed quality and about the varietal identity of the seeds have commonly led to seed laws. However, formal regulations are often inappropriate for informal seed systems, which

The beginning of a seed: regulatory mechanisms of double fertilization.

Science.gov (United States)

Bleckmann, Andrea; Alter, Svenja; Dresselhaus, Thomas

2014-01-01

THE LAUNCH OF SEED DEVELOPMENT IN FLOWERING PLANTS (ANGIOSPERMS) IS INITIATED BY THE PROCESS OF DOUBLE FERTILIZATION: two male gametes (sperm cells) fuse with two female gametes (egg and central cell) to form the precursor cells of the two major seed components, the embryo and endosperm, respectively. The immobile sperm cells are delivered by the pollen tube toward the ovule harboring the female gametophyte by species-specific pollen tube guidance and attraction mechanisms. After pollen tube burst inside the female gametophyte, the two sperm cells fuse with the egg and central cell initiating seed development. The fertilized central cell forms the endosperm while the fertilized egg cell, the zygote, will form the actual embryo and suspensor. The latter structure connects the embryo with the sporophytic maternal tissues of the developing seed. The underlying mechanisms of double fertilization are tightly regulated to ensure delivery of functional sperm cells and the formation of both, a functional zygote and endosperm. In this review we will discuss the current state of knowledge about the processes of directed pollen tube growth and its communication with the synergid cells resulting in pollen tube burst, the interaction of the four gametes leading to cell fusion and finally discuss mechanisms how flowering plants prevent multiple sperm cell entry (polyspermy) to maximize their reproductive success.

Seed characters and their usefulness in the separation of Asteraceae species

Directory of Open Access Journals (Sweden)

Michelli Fernandes Batista

2015-10-01

Full Text Available Studies on seed structure of Asteraceae have received little attention of botanists. Seed structure in Asteraceae is analyzed to evaluate the usefulness of characters in the separation of species through the analysis of nine weedy species. Seeds originate from anatropous, unitegmic and tenuinucellate ovules. The partial collapse of the developing seed coat is a common characteristic in the Asteraceae species studied. The testa consists of crushed and thin-walled cells in almost all of the species studied herein, except for Elephantopus mollis and Parthenium hysterophorus, which showed exotesta cells with U-shaped thickening. The analysis revealed high uniformity in seed characters.

Antiproliferative effect of isolated isoquinoline alkaloid from Mucuna pruriens seeds in hepatic carcinoma cells.

Science.gov (United States)

Kumar, Pranesh; Rawat, Atul; Keshari, Amit K; Singh, Ashok K; Maity, Siddhartha; De, Arnab; Samanta, Amalesh; Saha, Sudipta

2016-01-01

The present study was undertaken to investigate the antiproliferative action of isolated M1 (6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid) from Mucuna pruriens seeds using human hepatic carcinoma cell line (Huh-7 cells). Initially, docking studies was performed to find out the binding affinities of M1 to caspase-3 and 8 enzymes. Later, cytotoxic action of M1 was measured by cell growth inhibition (MTT), followed by caspase-3 and 8 enzymes assay colorimetrically. Our results collectively suggested that M1 had strong binding affinity to caspase-8 in molecular modelling. M1 possessed antiproliferative activity on Huh-7 cells (EC50 = 13.97 μM) and also inhibited the action of caspase-8 enzyme, signified process of apoptosis. M1 was active against Huh-7 cells that may be useful for future hepatic cancer treatment.

Perfusion seed cultures improve biopharmaceutical fed-batch production capacity and product quality.

Science.gov (United States)

Yang, William C; Lu, Jiuyi; Kwiatkowski, Chris; Yuan, Hang; Kshirsagar, Rashmi; Ryll, Thomas; Huang, Yao-Ming

2014-01-01

Volumetric productivity and product quality are two key performance indicators for any biopharmaceutical cell culture process. In this work, we showed proof-of-concept for improving both through the use of alternating tangential flow perfusion seed cultures coupled with high-seed fed-batch production cultures. First, we optimized the perfusion N-1 stage, the seed train bioreactor stage immediately prior to the production bioreactor stage, to minimize the consumption of perfusion media for one CHO cell line and then successfully applied the optimized perfusion process to a different CHO cell line. Exponential growth was observed throughout the N-1 duration, reaching >40 × 10(6) vc/mL at the end of the perfusion N-1 stage. The cultures were subsequently split into high-seed (10 × 10(6) vc/mL) fed-batch production cultures. This strategy significantly shortened the culture duration. The high-seed fed-batch production processes for cell lines A and B reached 5 g/L titer in 12 days, while their respective low-seed processes reached the same titer in 17 days. The shortened production culture duration potentially generates a 30% increase in manufacturing capacity while yielding comparable product quality. When perfusion N-1 and high-seed fed-batch production were applied to cell line C, higher levels of the active protein were obtained, compared to the low-seed process. This, combined with correspondingly lower levels of the inactive species, can enhance the overall process yield for the active species. Using three different CHO cell lines, we showed that perfusion seed cultures can optimize capacity utilization and improve process efficiency by increasing volumetric productivity while maintaining or improving product quality. © 2014 American Institute of Chemical Engineers.

The use of flow cytometry to accurately ascertain total and viable counts of Lactobacillus rhamnosus in chocolate.

Science.gov (United States)

Raymond, Yves; Champagne, Claude P

2015-04-01

The goals of this study were to evaluate the precision and accuracy of flow cytometry (FC) methodologies in the evaluation of populations of probiotic bacteria (Lactobacillus rhamnosus R0011) in two commercial dried forms, and ascertain the challenges in enumerating them in a chocolate matrix. FC analyses of total (FC(T)) and viable (FC(V)) counts in liquid or dried cultures were almost two times more precise (reproducible) than traditional direct microscopic counts (DCM) or colony forming units (CFU). With FC, it was possible to ascertain low levels of dead cells (FC(D)) in fresh cultures, which is not possible with traditional CFU and DMC methodologies. There was no interference of chocolate solids on FC counts of probiotics when inoculation was above 10(7) bacteria per g. Addition of probiotics in chocolate at 40 °C resulted in a 37% loss in viable cells. Blending of the probiotic powder into chocolate was not uniform which raised a concern that the precision of viable counts could suffer. FCT data can serve to identify the correct inoculation level of a sample, and viable counts (FCV or CFU) can subsequently be better interpreted. Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

Anticancer Screening of Various Seed Extract of Cardiospermum halicacabum on Human Colorectal, Skin and Breast Cancer Cell Lines

Directory of Open Access Journals (Sweden)

Behzad Mohaddesi

2015-08-01

Full Text Available Background: In the modern lifestyle, the increase in cancer and related chronic disorders is a major public health problem. In spite of different methods used for the treatment of these conditions, natural medicines have high demands due to their significant effects as immune enhancement and therapeutic agents and fewer side effects in comparison with other treatment methods. Hence, this study was undertaken to evaluate the cytotoxic effect of cardiospermum halicacabum Linn. seeds, based on traditional claims.Methods: A Soxhlet extractor was used to obtain different extracts from seeds of C. halicacabum Linn. Sulforhodamine B colorimetric (SRB assay used for the evaluation of the cytotoxic effect of the various extracts on HT-29, HCT-15 colon carcinoma, SK-MEL-2 skin carcinoma, and MCF-7 breast carcinoma. The results were compared against Doxorubicin as a standard drug.Results: The results of the present study showed the potent cytotoxic activity of n-hexane extract of seeds of C. halicacabum Linn. against the MCF-7 breast cancer cell line with 50% growth inhibition value (GI50 of 12.8 μg/ml but other extracts showed poor activity in other tested cell lines.Conclusions: The results indicated the potential medicinal value of C. halicacabum Linn. seeds oil with the highest extractive yield as an antineoplastic agent. However, further studies are needed for the isolation of the active anticancer compounds and evaluating the mechanism of action of the responsible compound.

AUTOLOGOUS Marrow-Derived Stem Cell-Seeded Gene-Supplemented Collagen Scaffolds for Spinal Cord Regeneration as a Treatment for Paralysis

National Research Council Canada - National Science Library

Spector, Myron

2006-01-01

.... Moreover, the authors will be investigating the effects of incorporating genes from nerve growth factors into the collagen scaffolds and seeding the scaffolds with marrow-derived mesenchymal stem cells...

Germination and soil seed bank traits of Podocarpus angustifolius (Podocarpaceae): an endemic tree species from Cuban rain forests.

Science.gov (United States)

Ferrandis, Pablo; Bonilla, Marta; Osorio, Licet del Carmen

2011-09-01

Podocarpus angustifolius is an endangered recalcitrant-seeded small tree, endemic to mountain rain forests in the central and Pinar del Río regions in Cuba. In this study, the germination patterns of P. angustifolius seeds were evaluated and the nature of the soil seed bank was determined. Using a weighted two-factor design, we analyzed the combined germination response to seed source (i.e. freshly matured seeds directly collected from trees versus seeds extracted from soil samples) and pretreatment (i.e. seed water-immersion for 48h at room temperature). Germination was delayed for four weeks (= 30 days) in all cases, regardless of both factors analyzed. Moreover, nine additional days were necessary to achieve high germination values (in the case of fresh, pretreated seeds). These results overall may indicate the existence of a non-deep simple morphophysiological dormancy in P. angustifolius seeds. The water-immersion significantly enhanced seed germination, probably as a result of the hydration of recalcitrant seeds. Although germination of seeds extracted from soil samples was low, probably due to aging and pathogen effects throughout the time of burial, the study revealed the existence of a persistent soil seed bank (at least short-termed) of approximately 42 viable seeds per m2 in the upper 10cm of soil. Such a record is noteworthy since references to persistent soil seed banks in recalcitrant-seeded species are scarce in the literature. The population consequences derived from the formation of persistent soil seed banks in this endangered species are discussed.

Rapid cell separation with minimal manipulation for autologous cell therapies

Science.gov (United States)

Smith, Alban J.; O'Rorke, Richard D.; Kale, Akshay; Rimsa, Roberts; Tomlinson, Matthew J.; Kirkham, Jennifer; Davies, A. Giles; Wälti, Christoph; Wood, Christopher D.

2017-02-01

The ability to isolate specific, viable cell populations from mixed ensembles with minimal manipulation and within intra-operative time would provide significant advantages for autologous, cell-based therapies in regenerative medicine. Current cell-enrichment technologies are either slow, lack specificity and/or require labelling. Thus a rapid, label-free separation technology that does not affect cell functionality, viability or phenotype is highly desirable. Here, we demonstrate separation of viable from non-viable human stromal cells using remote dielectrophoresis, in which an electric field is coupled into a microfluidic channel using shear-horizontal surface acoustic waves, producing an array of virtual electrodes within the channel. This allows high-throughput dielectrophoretic cell separation in high conductivity, physiological-like fluids, overcoming the limitations of conventional dielectrophoresis. We demonstrate viable/non-viable separation efficacy of >98% in pre-purified mesenchymal stromal cells, extracted from human dental pulp, with no adverse effects on cell viability, or on their subsequent osteogenic capabilities.

A non-invasive and rapid seed vigor biosensor based on quantitative measurement of superoxide generated by aleurone cell in intact seeds.

Science.gov (United States)

Liu, Xuejun; Gao, Caiji; Xing, Da

2009-02-15

Superoxide generated during the early imbibition is an excellent marker for evaluating seed vigor. In this paper, a new principle biosensor for non-invasive detection of seed vigor based on quantitative measurement of superoxide via selective probe 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo [1,2alpha] pyrazin-3-one (MCLA)-mediated chemiluminescence (CL) was developed. The biosensor, which used a compact single-photon counting module (SPCM) to collect the CL signal, could evaluate seed vigor in vivo. Benefiting from the high CL efficiency of MCLA reacting with superoxide and high sensitivity of the SPCM technique, the trace superoxide generated by dry seeds under storage state can be detected to achieve rapid and non-invasive determination of the seed vigor. In comparison with the traditional methods for fast measuring seed vigor based on measurement of physiological and biochemical properties, our proposed technique has significant advantages such as low cost, simplicity, convenient operation and short time consuming. To demonstrate the utility of the system, it was applied to evaluate MCLA-mediated CL of three different plant species wheat (Ze Yu No. 2), maize (Tai Gu No. 1 and 2) and rice (Jing Dao No. 21) seeds with different degrees of aging. The experimental results suggested that there was an excellent positive correlation between the seed vigor assessment from quantitative TTC-test and the detection based on MCLA-mediated CL of superoxide measurement. The new principle of seed vigor measurement is a challenge and breakthrough to conventional method of seed vigor determination and may be a potential technique of the next generation seed vigor detection.

Development of maternal seed tissue in barley is mediated by regulated cell expansion and cell disintegration and coordinated with endosperm growth.

Science.gov (United States)

Radchuk, Volodymyr; Weier, Diana; Radchuk, Ruslana; Weschke, Winfriede; Weber, Hans

2011-01-01

After fertilization, filial grain organs are surrounded by the maternal nucellus embedded within the integuments and pericarp. Rapid early endosperm growth must be coordinated with maternal tissue development. Parameters of maternal tissue growth and development were analysed during early endosperm formation. In the pericarp, cell proliferation is accomplished around the time of fertilization, followed by cell elongation predominantly in longitudinal directions. The rapid cell expansion coincides with endosperm cellularization. Distribution of TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling)-positive nuclei reveals distinct patterns starting in the nucellus at anthesis and followed later by the inner cell rows of the pericarp, then spreading to the whole pericarp. The pattern suggests timely and spatially regulated programmed cell death (PCD) processes in maternal seed tissues. When the endosperm is coenocytic, PCD events are only observed within the nucellus. Thereby, remobilization of nucellar storage compounds by PCD could nourish the early developing endosperm when functional interconnections are absent between maternal and filial seed organs. Specific proteases promote PCD events. Characterization of the barley vacuolar processing enzyme (VPE) gene family identified seven gene members specifically expressed in the developing grain. HvVPE2a (known as nucellain) together with closely similar HvVPE2b and HvVPE2d might be involved in nucellar PCD. HvVPE4 is strongly cell specific for pericarp parenchyma. Correlative evidence suggests that HvVPE4 plays a role in PCD events in the pericarp. Possible functions of PCD in the maternal tissues imply a potential nutritive role or the relief of a physical restraint for endosperm growth. PCD could also activate post-phloem transport functions.

DNA comet assay for rice seeds treated with low energy electrons ('soft-electrons')

International Nuclear Information System (INIS)

Todoriki, Setsuko; Hayashi, Toru

1999-01-01

As rice seeds are sometimes contaminated with phytopathogenic organisms such as blast disease fungi and nematodes, a novel non-chemical disinfection method for rice seeds is highly required. In order to develop a disinfection method, the effect of low energy electron ('soft-electrons') on seed DNA was examined by using the neutral comet assay. Rice seeds (whole grain) were treated with electrons of different acceleration voltages (180 kV to 1 MV) at a dose of 5 kGy. Nucleus suspensions were prepared from whole brown rice and subjected to electrophoresis. DNA from un-irradiated (control) seeds relaxed and produced comets with a short tail, most of the comets distributed within the range of comet length between 30 μm to 70 μm. In the case of seeds treated with electrons at acceleration voltages up to 190 kV, cells without seed coats were not damaged and the frequency histograms of comet length showed almost the same pattern as that for control. At acceleration voltages higher than 200 kV, the cells were distributed into two categories; DNA comets with a short tail (with little DNA damages, less than 70 μm in the comet length) and DNA comets with long tails (with sever strand breaks, more than 130 μm in the comet length). The ratios of damaged cells increased with increasing acceleration voltage. The growths of rice seedlings were not affected by the treatment with electrons at up to 200 kV. On the contrary, the cells of gamma-irradiated seed showed small variations in the comet length, and which were depending on radiation dose. The individual cells of gamma-irradiated seeds at 1 kGy showed shorter comet than the damaged cells with soft electron, seed treated with gamma rays (1-5 kGy) did not shoot nor root. (author)

SEED INOCULATION WITH Azospirillum brasilense, ASSOCIATED WITH THE USE OF BIOREGULATORS IN MAIZE

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ALESSANDRO DE LUCCA E BRACCINI

2012-01-01

Full Text Available The inoculation of seeds with the bacterium Azospirillum has been carried out in maize culture and other grasses. The application of growth bio-regulators is another technology whose results in maize culture have yet to become more widespread. Current study evaluates the agronomic effectiveness of seed inoculation with Azospirillum brasilense in maize, associated with the use of the growth regulator Stimulate ®. Triple hybrid maize CD 304 underwent the following treatments: 1 - control without nitrogen and without Azospirillum brasilense; 2 - Treatment without nitrogen but with Azospirillum brasilense; 3 - Treatment without nitrogen but with Azospirillum brasilense + Stimulate ®; 4 - Treatment with 50% of nitrogen dose recommended for maize culture; 5 - Treatment with 50% of nitrogen dose and inoculation with Azospirillum brasilense; 6 - Same as 5 but with Stimulate ®; 7 - Total N recommended; 8 - Total N recommended + Azospirillum brasilense ; 9 - Total N recommended + Azospirillum brasilense + Stimulate ®. The inoculation of maize seeds with Azospirillum brasilense increases plant height and grain yield when compared with rates in control. The use of 50% of N dose in sowing, associated with the inoculation of maize seeds with Azospirillum brasilense at 200 mL ha-1 (mixed to the seeds and associated with Stimulate ® (in foliar application, is viable.

In Situ Spatiotemporal Mapping of Flow Fields around Seeded Stem Cells at the Subcellular Length Scale

Science.gov (United States)

Song, Min Jae; Dean, David; Knothe Tate, Melissa L.

2010-01-01

A major hurdle to understanding and exploiting interactions between the stem cell and its environment is the lack of a tool for precise delivery of mechanical cues concomitant to observing sub-cellular adaptation of structure. These studies demonstrate the use of microscale particle image velocimetry (μ-PIV) for in situ spatiotemporal mapping of flow fields around mesenchymal stem cells, i.e. murine embryonic multipotent cell line C3H10T1/2, at the subcellular length scale, providing a tool for real time observation and analysis of stem cell adaptation to the prevailing mechanical milieu. In the absence of cells, computational fluid dynamics (CFD) predicts flow regimes within 12% of μ-PIV measures, achieving the technical specifications of the chamber and the flow rates necessary to deliver target shear stresses at a particular height from the base of the flow chamber. However, our μ-PIV studies show that the presence of cells per se as well as the density at which cells are seeded significantly influences local flow fields. Furthermore, for any given cell or cell seeding density, flow regimes vary significantly along the vertical profile of the cell. Hence, the mechanical milieu of the stem cell exposed to shape changing shear stresses, induced by fluid drag, varies with respect to proximity of surrounding cells as well as with respect to apical height. The current study addresses a previously unmet need to predict and observe both flow regimes as well as mechanoadaptation of cells in flow chambers designed to deliver precisely controlled mechanical signals to live cells. An understanding of interactions and adaptation in response to forces at the interface between the surface of the cell and its immediate local environment may be key for de novo engineering of functional tissues from stem cell templates as well as for unraveling the mechanisms underlying multiscale development, growth and adaptation of organisms. PMID:20862249

In situ spatiotemporal mapping of flow fields around seeded stem cells at the subcellular length scale.

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Min Jae Song

2010-09-01

Full Text Available A major hurdle to understanding and exploiting interactions between the stem cell and its environment is the lack of a tool for precise delivery of mechanical cues concomitant to observing sub-cellular adaptation of structure. These studies demonstrate the use of microscale particle image velocimetry (μ-PIV for in situ spatiotemporal mapping of flow fields around mesenchymal stem cells, i.e. murine embryonic multipotent cell line C3H10T1/2, at the subcellular length scale, providing a tool for real time observation and analysis of stem cell adaptation to the prevailing mechanical milieu. In the absence of cells, computational fluid dynamics (CFD predicts flow regimes within 12% of μ-PIV measures, achieving the technical specifications of the chamber and the flow rates necessary to deliver target shear stresses at a particular height from the base of the flow chamber. However, our μ-PIV studies show that the presence of cells per se as well as the density at which cells are seeded significantly influences local flow fields. Furthermore, for any given cell or cell seeding density, flow regimes vary significantly along the vertical profile of the cell. Hence, the mechanical milieu of the stem cell exposed to shape changing shear stresses, induced by fluid drag, varies with respect to proximity of surrounding cells as well as with respect to apical height. The current study addresses a previously unmet need to predict and observe both flow regimes as well as mechanoadaptation of cells in flow chambers designed to deliver precisely controlled mechanical signals to live cells. An understanding of interactions and adaptation in response to forces at the interface between the surface of the cell and its immediate local environment may be key for de novo engineering of functional tissues from stem cell templates as well as for unraveling the mechanisms underlying multiscale development, growth and adaptation of organisms.

A Spectrophotometric Assay for Robust Viability Testing of Seed Batches Using 2,3,5-Triphenyl Tetrazolium Chloride: Using Hordeum vulgare L. as a Model

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Laura Lopez Del Egido

2017-05-01

Full Text Available A comparative analysis was carried out of published methods to assess seed viability using 2,3,5-triphenyltetrazolium chloride (TTC based assays of seed batches. The tests were carried out on seeds of barley (Hordeum vulgare cv. Optic as a model. We established that 10% [w/v] trichloroacetic acid (TCA/methanol is superior to the acetone and methanol-only based methods: allowing the highest recovery of formazan and the lowest background optical density (OD readings, across seed lots comprising different ratios of viable and dead seeds. The method allowed a linear-model to accurately capture the statistically significant relationship between the quantity of formazan that could be extracted using the method we developed and the seed temperature-response, and seed viability as a function of artificially aged seed lots. Other quality control steps are defined to help ensure the assay is robust and these are reported in a Standard Operating Procedure.

Detection of viable Salmonella in ice cream by TaqMan real-time polymerase chain reaction assay combining propidium monoazide

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Yuexia Wang

2015-09-01

Full Text Available Real-time polymerase chain reaction (PCR allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at −18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 103 CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 100 CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach.

Detection of viable Salmonella in ice cream by TaqMan real-time polymerase chain reaction assay combining propidium monoazide.

Science.gov (United States)

Wang, Yuexia; Yang, Ming; Liu, Shuchun; Chen, Wanyi; Suo, Biao

2015-09-01

Real-time polymerase chain reaction (PCR) allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at -18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA) was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 10 3  CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 10 0  CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach. Copyright © 2015. Published by Elsevier B.V.

Molecular identification of broomrape species from a single seed by High Resolution Melting analysis

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Mathieu Rolland

2016-12-01

Full Text Available Broomrapes are holoparasitic plants spreading through seeds. Each plant produces hundreds of thousands of seeds which remain viable in the soils for decades. To limit their spread, drastic measures are being taken and the contamination of a commercial seed lot by a single broomrape seed can lead to its rejection. Considering that broomrapes species identification from a single seed is extremely difficult even for trained botanists and that among all the described species, only a few are really noxious for the crops, numerous seed lots are rejected because of the contamination by seeds of non-noxious broomrape species. The aim of this study was to develop and evaluate a High Resolution Melting assay identifying the eight most noxious and common broomrape species (P. aegyptiaca, O. cernua, O. crenata, O. cumana, O. foetida, O. hederae, O. minor, and P. ramosa from a single seed. Based on trnL and rbcL plastidial genes amplification, the designed assay successfully identifies O. cumana, O. cernua, O. crenata, O. minor, O. hederae, and O. foetida; P. ramosa and P. aegyptiaca can be differentiated from other species but not from each other. Tested on 50 seed lots, obtained results perfectly matched identifications performed by sequencing. Through the analysis of common seed lots by different analysts, the reproducibility of the assay was evaluated at 90 %. Despite an original sample preparation process it was not possible to extract enough DNA from some seeds (10% of the samples. The described assay fulfils its objectives and allows an accurate identification of the targeted broomrape species. It can be used to identify contaminants in commercial seed lots or for any other purpose. The assay might be extended to vegetative material.

Morpho-anatomy, imbibition, viability and germination of the seed of Anadenanthera colubrina var. cebil (Fabaceae).

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Varela, Rodolfo Omar; Albornoz, Patricia Liliana

2013-09-01

Seed biology is a relevant aspect of tropical forests because it is central to the understanding of processes of plant establishment, succession and natural regeneration. Anadenanthera colubrina var. cebil is a timber tree from South America that produces large seeds with thin weak teguments, which is uncommon among legumes. This study describes the morphology and anatomy of the seed coat, the viability, imbibition, and germination in this species. Seeds used during the essays came from 10 trees that grow naturally in Horco Molle, province of Tucumán, Argentina. Seed morphology was described from a sample of 20 units. The seed coat surface was examined with a scanning electron microscope. Transverse sections of hydrated and non-hydrated seeds were employed to describe the histological structure of the seed coat. Hydration, viability and germination experiments were performed under laboratory controlled conditions; and the experimental design consisted of 10 replicas of 10 seeds each. Viability and germination tests were conducted using freshly fallen seeds and seeds stored for five months. Morphologically the seeds of A. colubrina var. cebil are circular to subcircular, laterally compressed, smooth, bright brown and have a horseshoe fissure line (= pleurogram) on both sides. The seed coat comprises five tissue layers and a double (external and internal) cuticle. The outer cuticle (on the epidermis) is smooth and interrupted by microcracks and pores of variable depth. The epidermis consists of macroesclereids with non-lignified secondary walls. This layer is separated from the underlying ones during seed hydration. The other layers of internal tissues are comprised of osteosclereids, parenchyma, osteosclereids, and macrosclereids. The percentage of viable seeds was 93%, decreasing to 75% in seeds with five months old. Seed mass increased 76% after the first eight hours of hydration. Germination percentage was 75% after 76 hours. Germination of seeds stored for five

Enhancement of local species richness in tundra by seed dispersal through guts of muskox and barnacle goose

DEFF Research Database (Denmark)

Bruun, Hans Henrik; Lundgren, Rebekka; Philipp, Marianne

2008-01-01

The potential contribution of vertebrate-mediated seed rain to the maintenance of plant community richness in a High Arctic ecosystem was investigated. We analyzed viable seed content in dung of the four numerically most important terrestrial vertebrates in Northeast Greenland - muskox (Ovibos...... moschatus), barnacle goose (Branta leucopsis), Arctic fox (Alopex lagopus) and Arctic hare (Lepus arcticus). High numbers of plant propagules were found in the dung of muskox and barnacle goose. Seeds of many plant species were found in the faeces of one vertebrate species only. Propagule composition...... in barnacle goose droppings was relatively uniform over samples, with a high abundance of the nutritious bulbils of Polygonum viviparum (Bistorta vivipara), suggesting that geese have a narrow habitat preference and feed selectively. Propagule composition in muskox dung was diverse and heterogeneous among...

Detection and Quantification of Viable and Nonviable Trypanosoma cruzi Parasites by a Propidium Monoazide Real-Time Polymerase Chain Reaction Assay

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Cancino-Faure, Beatriz; Fisa, Roser; Alcover, M. Magdalena; Jimenez-Marco, Teresa; Riera, Cristina

2016-01-01

Molecular techniques based on real-time polymerase chain reaction (qPCR) allow the detection and quantification of DNA but are unable to distinguish between signals from dead or live cells. Because of the lack of simple techniques to differentiate between viable and nonviable cells, the aim of this study was to optimize and evaluate a straightforward test based on propidium monoazide (PMA) dye action combined with a qPCR assay (PMA-qPCR) for the selective quantification of viable/nonviable epimastigotes of Trypanosoma cruzi. PMA has the ability to penetrate the plasma membrane of dead cells and covalently cross-link to the DNA during exposure to bright visible light, thereby inhibiting PCR amplification. Different concentrations of PMA (50–200 μM) and epimastigotes of the Maracay strain of T. cruzi (1 × 105–10 parasites/mL) were assayed; viable and nonviable parasites were tested and quantified by qPCR with a TaqMan probe specific for T. cruzi. In the PMA-qPCR assay optimized at 100 μM PMA, a significant qPCR signal reduction was observed in the nonviable versus viable epimastigotes treated with PMA, with a mean signal reduction of 2.5 logarithm units and a percentage of signal reduction > 98%, in all concentrations of parasites assayed. This signal reduction was also observed when PMA-qPCR was applied to a mixture of live/dead parasites, which allowed the detection of live cells, except when the concentration of live parasites was low (10 parasites/mL). The PMA-qPCR developed allows differentiation between viable and nonviable epimastigotes of T. cruzi and could thus be a potential method of parasite viability assessment and quantification. PMID:27139452

Frugivory and seed dispersal by birds in Cereus jamacaru DC. ssp. jamacaru (Cactaceae) in the Caatinga of Northeastern Brazil.

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Gomes, V G N; Quirino, Z G M; Araujo, H F P

2014-02-01

Studies of the dispersal modes of plants aid our understanding of the dynamics of resource and its availability for dispersal agents. The present work sought to characterize the fruiting patterns of the native Caatinga (dryland) cactus Cereus jamacaru, identify its principal dispersers, and evaluate the effects of seed passage through digestive tract of dispersers on its germination. Cereus jamacaru present an annual fruiting pattern and fruiting peaks occurred during June/2009 and February/2010. A total of 135 visits by nine species of resident Caatinga bird species were recorded. The most frequent visiting bird species were Paroaria dominicana and Euphonia chlorotica. Length of bird visits varied from 15 seconds to 4 minutes and seeds removed by birds travelled 10.6 ± 11.2 m until dispersers make the first landing perch, in some cases more than 40 meters away. Germination tests show birds had a high quantity of viable seeds of C. jamacaru in its feces. Seeds that passed through the digestive tract of birds showed a similar germinability of the seeds of the control group. However, the seeds dispersed by birds showed lowest mean germination time related to the control group seeds. This study highlights the potential role of birds as seed dispersers of C. jamacaru, swallowing the whole seeds and defecating intact seeds, accelerating the germination process and transporting seeds away from the mother plant.

Enumeration of viable and non-viable larvated Ascaris eggs with quantitative PCR

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Aims: The goal of the study was to further develop an incubation-qPCR method for quantifying viable Ascaris eggs. The specific objectives were to characterize the detection limit and number of template copies per egg, determine the specificity of the method, and test the method w...

Importance of earthworm-seed interactions for the composition and structure of plant communities: A review

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Forey, Estelle; Barot, Sébastien; Decaëns, Thibaud; Langlois, Estelle; Laossi, Kam-Rigne; Margerie, Pierre; Scheu, Stefan; Eisenhauer, Nico

2011-11-01

Soil seed bank composition and dynamics are crucial elements for the understanding of plant population and community ecology. Earthworms are increasingly recognized as important dispersers and predators of seeds. Through direct and indirect effects they influence either positively or negatively the establishment and survival of seeds and seedlings. Seedling establishment is affected by a variety of earthworm-mediated mechanisms, such as selective seed ingestion and digestion, acceleration or deceleration of germination, and seed transport. Earthworm casts deposited on the soil surface and the entrance of earthworm burrows often contain viable seeds and constitute important regeneration niches for plant seedlings and therefore likely favour specific seed traits. However, the role of earthworms as seed dispersers, mediators of seed bank dynamics and seed predators has not been considered in concert. The overall effect of earthworms on plant communities remains little understood. Most knowledge is based on laboratory studies on temperate species and future work has to explore the biological significance of earthworm-seed interactions under more natural conditions. In this review we summarize the current knowledge on earthworm-seed interactions and discuss factors determining these interactions. We highlight that this interaction may be an underappreciated, yet major driving force for the dynamics of soil seed banks and plant communities which most likely have experienced co-evolutionary processes. Despite the experimental bias, we hypothesize that the knowledge gathered in the present review is of crucial relevance for restoration and conservation ecology. For instance, as earthworms emerge as successful and ubiquitous invaders in various ecosystems, the summarized information might serve as a basis for realistic estimations and modelling of consequences on native plant communities. We depict promising directions of future research and point to the need to consider

Bone repair by cell-seeded 3D-bioplotted composite scaffolds made of collagen treated tricalciumphosphate or tricalciumphosphate-chitosan-collagen hydrogel or PLGA in ovine critical-sized calvarial defects.

Science.gov (United States)

Haberstroh, Kathrin; Ritter, Kathrin; Kuschnierz, Jens; Bormann, Kai-Hendrik; Kaps, Christian; Carvalho, Carlos; Mülhaupt, Rolf; Sittinger, Michael; Gellrich, Nils-Claudius

2010-05-01

The aim of this study was to investigate the osteogenic effect of three different cell-seeded 3D-bioplotted scaffolds in a ovine calvarial critical-size defect model. The choice of scaffold-materials was based on their applicability for 3D-bioplotting and respective possibility to produce tailor-made scaffolds for the use in cranio-facial surgery for the replacement of complex shaped boneparts. Scaffold raw-materials are known to be osteoinductive when being cell-seeded [poly(L-lactide-co-glycolide) (PLGA)] or having components with osteoinductive properties as tricalciumphosphate (TCP) or collagen (Col) or chitosan. The scaffold-materials PLGA, TCP/Col, and HYDR (TCP/Col/chitosan) were cell-seeded with osteoblast-like cells whether gained from bone (OLB) or from periost (OLP). In a prospective and randomized design nine sheep underwent osteotomy to create four critical-sized calvarial defects. Three animals each were assigned to the HYDR-, the TCP/Col-, or the PLGA-group. In each animal, one defect was treated with a cell-free, an OLB- or OLP-seeded group-specific scaffold, respectively. The fourth defect remained untreated as control (UD). Fourteen weeks later, animals were euthanized for histo-morphometrical analysis of the defect healing. OLB- and OLP-seeded HYDR and OLB-seeded TCP/Col scaffolds significantly increased the amount of newly formed bone (NFB) at the defect bottom and OLP-seeded HYDR also within the scaffold area, whereas PLGA-scaffolds showed lower rates. The relative density of NFB was markedly higher in the HYDR/OLB group compared to the corresponding PLGA group. TCP/Col had good stiffness to prepare complex structures by bioplotting but HYDR and PLGA were very soft. HYDR showed appropriate biodegradation, TCP/Col and PLGA seemed to be nearly undegraded after 14 weeks. 3D-bioplotted, cell-seeded HYDR and TCP/Col scaffolds increased the amount of NFB within ovine critical-size calvarial defects, but stiffness, respectively, biodegradation of

Internalisation and multiple phosphorylation of γ-Conglutin, the lupin seed glycaemia-lowering protein, in HepG2 cells

International Nuclear Information System (INIS)

Capraro, Jessica; Magni, Chiara; Faoro, Franco; Maffi, Dario; Scarafoni, Alessio; Tedeschi, Gabriella; Maffioli, Elisa; Parolari, Anna; Manzoni, Cristina; Lovati, Maria Rosa; Duranti, Marcello

2013-01-01

Highlights: •A glycaemia-reducing lupin seed protein is internalized by HepG2 cells. •The protein accumulates in the cytosol in an intact form. •The internalized protein is multiply phosphorylated. -- Abstract: Lupin seed γ-Conglutin is a protein capable of reducing glycaemia in mammalians and increasing glucose uptake by model cells. This work investigated whether γ-Conglutin is internalised into the target cells and undergoes any covalent change during the process, as a first step to understanding its mechanism of action. To this purpose, γ-Conglutin-treated and untreated HepG2 cells were submitted to confocal and transmission electron microscopy. Immune-revelation of γ-Conglutin at various intervals revealed its accumulation inside the cytosol. In parallel, 2D-electrophoresis of the cell lysates and antibody reaction of the blotted maps showed the presence of the protein intact subunits inside the treated cells, whilest no trace of the protein was found in the control cells. However, γ-Conglutin-related spots with an unexpectedly low pI were also observed in the maps. These spots were excised, trypsin-treated and submitted to MS/MS spectrometric analysis. The presence of phosphorylated amino acids was detected. These findings, by showing that γ-Conglutin is internalised by HepG2 cells in an intact form and is modified by multiple phosphorylation, open the way to the understanding of the lupin γ-Conglutin insulin-mimetic activity

Auxin production in the endosperm drives seed coat development in Arabidopsis

Science.gov (United States)

Figueiredo, Duarte D; Batista, Rita A; Roszak, Pawel J; Hennig, Lars; Köhler, Claudia

2016-01-01

In flowering plants, seed development is initiated by the fusion of the maternal egg and central cells with two paternal sperm cells, leading to the formation of embryo and endosperm, respectively. The fertilization products are surrounded by the maternally derived seed coat, whose development prior to fertilization is blocked by epigenetic regulators belonging to the Polycomb Group (PcG) protein family. Here we show that fertilization of the central cell results in the production of auxin and most likely its export to the maternal tissues, which drives seed coat development by removing PcG function. We furthermore show that mutants for the MADS-box transcription factor AGL62 have an impaired transport of auxin from the endosperm to the integuments, which results in seed abortion. We propose that AGL62 regulates auxin transport from the endosperm to the integuments, leading to the removal of the PcG block on seed coat development. DOI: http://dx.doi.org/10.7554/eLife.20542.001 PMID:27848912

Repeated drought alters resistance of seed bank regeneration in baldcypress swamps of North America

Science.gov (United States)

Lei, Ting; Middleton, Beth A.

2018-01-01

Recurring drying and wetting events are likely to increase in frequency and intensity in predicted future droughts in the central USA and alter the regeneration potential of species. We explored the resistance of seed banks to successive droughts in 53 sites across the nine locations in baldcypress swamps in the southeastern USA. Along the Mississippi River Alluvial Valley and northern Gulf of Mexico, we investigated the capacity of seed banks to retain viable seeds after successive periods of drying and wetting in a greenhouse study. Mean differences in species richness and seed density were compared to examine the interactions of successive droughts, geographical location and water regime. The results showed that both species richness and total density of germinating seedlings decreased over repeated drought trials. These responses were more pronounced in geographical areas with higher annual mean temperature. In seed banks across the southeastern swamp region, most species were exhausted after Trial 2 or 3, except for semiaquatic species in Illinois and Tennessee, and aquatic species in Texas. Distinct geographical trends in seed bank resistance to drought demonstrate that climate-induced drying of baldcypress swamps could influence the regeneration of species differently across their ranges. Despite the health of adult individuals, lack of regeneration may push ecosystems into a relict status. Seed bank depletion by germination without replenishment may be a major conservation threat in a future with recurring droughts far less severe than megadrought. Nevertheless, the protection of moist refugia might aid conservation.

Germination and soil seed bank traits of Podocarpus angustifolius (Podocarpaceae: an endemic tree species from Cuban rain forests

Directory of Open Access Journals (Sweden)

Pablo Ferrandis

2011-09-01

Full Text Available Podocarpus angustifolius is an endangered recalcitrant-seeded small tree, endemic to mountain rain forests in the central and Pinar del Río regions in Cuba. In this study, the germination patterns of P. angustifolius seeds were evaluated and the nature of the soil seed bank was determined. Using a weighted two-factor design, we analyzed the combined germination response to seed source (i.e. freshly matured seeds directly collected from trees versus seeds extracted from soil samples and pretreatment (i.e. seed water-immersion for 48h at room temperature. Germination was delayed for four weeks (≈30 days in all cases, regardless of both factors analyzed. Moreover, nine additional days were necessary to achieve high germination values (in the case of fresh, pretreated seeds. These results overall may indicate the existence of a non-deep simple morphophysiological dormancy in P. angustifolius seeds. The water-immersion significantly enhanced seed germination, probably as a result of the hydration of recalcitrant seeds. Although germination of seeds extracted from soil samples was low, probably due to aging and pathogen effects throughout the time of burial, the study revealed the existence of a persistent soil seed bank (at least short-termed of ≈42 viable seeds per m² in the upper 10cm of soil. Such a record is noteworthy since references to persistent soil seed banks in recalcitrant-seeded species are scarce in the literature. The population consequences derived from the formation of persistent soil seed banks in this endangered species are discussed. Rev. Biol. Trop. 59 (3: 1061-1069. Epub 2011 September 01.

The antifungal properties of a 2S albumin-homologous protein from passion fruit seeds involve plasma membrane permeabilization and ultrastructural alterations in yeast cells.

Science.gov (United States)

Agizzio, Ana Paula; Da Cunha, Maura; Carvalho, André O; Oliveira, Marco Antônio; Ribeiro, Suzanna F F; Gomes, Valdirene M

2006-10-01

Different types of antimicrobial proteins were purified from plant seeds, including chitinases, β-1,3-glucanases, defensins, thionins, lipid transfer proteins and 2S albumins. It has become clear that these groups of proteins play an important role in the protection of plants from microbial infection. Recent results from our laboratory have shown that the defense-related proteins from passion fruit seeds, named Pf1 and Pf2 (which show sequence homology with 2S albumins), inhibit fungal growth and glucose-stimulated acidification of the medium by Saccharomyces cerevisiae cells. The aim of this study was to determine whether 2S albumins from passion fruit seeds induce plasma membrane permeabilization and cause morphological alterations in yeast cells. Initially, we used an assay based on the uptake of SYTOX Green, an organic compound that fluoresces upon interaction with nucleic acids and penetrates cells with compromised plasma membranes, to investigate membrane permeabilization in S. cerevisiae cells. When viewed with a confocal laser microscope, S. cervisiae cells showed strong SYTOX Green fluorescence in the cytosol, especially in the nuclei. 2S albumins also inhibited glucose-stimulated acidification of the medium by S. cerevisiae cells, which indicates a probable impairment of fungal metabolism. The microscopical analysis of the yeast cells treated with 2S albumins demonstrated several morphological alterations in cell shape, cell surface, cell wall and bud formation, as well as in the organization of intracellular organelles. Copyright © 2006 Elsevier Ireland Ltd. All rights reserved.

Local evolution of seed flotation in Arabidopsis.

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Susana Saez-Aguayo

2014-03-01

Full Text Available Arabidopsis seeds rapidly release hydrophilic polysaccharides from the seed coat on imbibition. These form a heavy mucilage layer around the seed that makes it sink in water. Fourteen natural Arabidopsis variants from central Asia and Scandinavia were identified with seeds that have modified mucilage release and float. Four of these have a novel mucilage phenotype with almost none of the released mucilage adhering to the seed and the absence of cellulose microfibrils. Mucilage release was modified in the variants by ten independent causal mutations in four different loci. Seven distinct mutations affected one locus, coding the MUM2 β-D-galactosidase, and represent a striking example of allelic heterogeneity. The modification of mucilage release has thus evolved a number of times independently in two restricted geographical zones. All the natural mutants identified still accumulated mucilage polysaccharides in seed coat epidermal cells. Using nuclear magnetic resonance (NMR relaxometry their production and retention was shown to reduce water mobility into internal seed tissues during imbibition, which would help to maintain seed buoyancy. Surprisingly, despite released mucilage being an excellent hydrogel it did not increase the rate of water uptake by internal seed tissues and is more likely to play a role in retaining water around the seed.

Glyoxal Crosslinking of Cell-Seeded Chitosan/Collagen Hydrogels for Bone Regeneration

Science.gov (United States)

Wang, Limin; Stegemann, Jan P.

2011-01-01

Chitosan and collagen are natural biomaterials that have been used extensively in tissue engineering, both separately and as composite materials. Most methods to fabricate chitosan/collagen composites use freeze drying and chemical crosslinking to create stable porous scaffolds, which subsequently can be seeded with cells. In this study, we directly embedded human bone marrow stem cells (hBMSC) in chitosan/collagen materials by initiating gelation using β-glycerophosphate at physiological temperature and pH. We further examined the use of glyoxal, a dialdehyde with relatively low toxicity, to crosslink these materials and characterized the resulting changes in matrix and cell properties. The cytocompatibility of glyoxal and the crosslinked gels were investigated in terms of hBMSC metabolic activity, viability, proliferation, and osteogenic differentiation. These studies revealed that glyoxal was cytocompatible at concentrations below about 1 mM for periods of exposure up to 15 h, though the degree of cell spreading and proliferation were dependent on matrix composition. Glyoxal-crosslinked matrices were stiffer and compacted less than uncrosslinked controls. It was further demonstrated that hBMSC can attach and proliferate in 3D matrices composed of 50/50 chitosan/collagen, and that these materials supported osteogenic differentiation in response to stimulation. Such glyoxal-crosslinked chitosan/collagen composite materials may find utility as cell delivery vehicles for enhancing the repair of bone defects. PMID:21345389

Effects of gamma radiations with or without chemical mutagens on rose seeds

International Nuclear Information System (INIS)

Lata, P.

1988-01-01

A preliminary experiment with seeds from 4 diploids, 17 tetraploids, 2 hexaploids and 1 octoploid roses confirmed that seed collection during early autumn under climatical conditions of London, gives a high percentage of viable seeds than those collected during late autumn. Seed stratification for about 100 days enhances germination when carried out under wet, purified sand. Harmful effects of radiations were recorded on seed germination, seedling survival, growth and flowering, after 0-120 Kr exposures of gamma rays. Rose cultivars were found to be more sensitive to radiations than the Rosa spp. The LD50 for diploid R. rugosa typica was between 20-30 Kr and for allo-tetraploid cultivar Aunty Dora, it was 8 Kr. No significant effect of radiations on seedling growth was recorded. Flowering in seedlings, raised from cultivar seeds, produced flowers during the first year of germination as compared with R. spp. seedlings which flowered after 3 to 4 years of germination. Among the chemical mutagens, Hydroxylamine produced least harmful effects on seed germination and survivals than those produced by Ethyl methane sulphonate; used alone or combined with low or high doses of radiations. The average seedling heights and number of leaves per seedlings were not affected. Mutagens were responsible for bringing about a loss in colour intensity which was more frequent when radiations and mutagens were used together. Real flower colour changes were rare and were attributed to the pollinating parent or mutagen or both. Present experiments indicate that although there are initial difficulties with seed germination but the resulting seedlings with increased genetic variability can generously reward a rose breeder in introducing new roses with rare combination of desirable characters which are difficult to accomplish merely by induced somatic mutations

Effects of pulsed magnetic field treatment of soybean seeds on calli growth, cell damage, and biochemical changes under salt stress.

Science.gov (United States)

Radhakrishnan, Ramalingam; Leelapriya, Thasari; Kumari, Bollipo Diana Ranjitha

2012-12-01

The effects of magnetic field (MF) treatments of soybean seeds on calli growth, cell damage, and biochemical changes under salt stress were investigated under controlled conditions. Soybean seeds were exposed to a 1.0 Hz sinusoidal uniform pulsed magnetic field (PMF) of 1.5 µT for 5 h/day for 20 days. Non-treated seeds were considered as controls. For callus regeneration, the embryonic axis explants were taken from seeds and inoculated in a saline medium with a concentration of 10 mM NaCl for calli growth analysis and biochemical changes. The combined treatment of MF and salt stress was found to significantly increase calli fresh weight, total soluble sugar, total protein, and total phenol contents, but it decreased the ascorbic acid, lipid peroxidation, and catalase activity of calli from magnetically exposed seeds compared to the control calli. PMF treatment significantly improved calli tolerance to salt stress in terms of an increase in flavonoid, flavone, flavonole, alkaloid, saponin, total polyphenol, genistein, and daidzein contents under salt stress. The results suggest that PMF treatment of soybean seeds has the potential to counteract the adverse effects of salt stress on calli growth by improving primary and secondary metabolites under salt stress conditions. Copyright © 2012 Wiley Periodicals, Inc.

High-Density Spot Seeding for Tissue Model Formation

Science.gov (United States)

Marquette, Michele L. (Inventor); Sognier, Marguerite A. (Inventor)

2016-01-01

A model of tissue is produced by steps comprising seeding cells at a selected concentration on a support to form a cell spot, incubating the cells to allow the cells to partially attach, rinsing the cells to remove any cells that have not partially attached, adding culture medium to enable the cells to proliferate at a periphery of the cell spot and to differentiate toward a center of the cell spot, and further incubating the cells to form the tissue. The cells may be C2C12 cells or other subclones of the C2 cell line, H9c2(2-1) cells, L6 cells, L8 cells, QM7 cells, Sol8 cells, G-7 cells, G-8 cells, other myoblast cells, cells from other tissues, or stem cells. The selected concentration is in a range from about 1 x 10(exp 5) cells/ml to about 1 x 10(exp 6) cells/ml. The tissue formed may be a muscle tissue or other tissue depending on the cells seeded.

Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture

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Restu Syamsul Hadi

2014-08-01

Full Text Available Background Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC. Methods In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4 expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1 proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test. Results Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05. Calculated viable cell numbers increased 1.8 and 5.1-fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity Conclusions Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.

Successful Isolation of Viable Adipose-Derived Stem Cells from Human Adipose Tissue Subject to Long-Term Cryopreservation: Positive Implications for Adult Stem Cell-Based Therapeutics in Patients of Advanced Age

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Sean M. Devitt

2015-01-01

Full Text Available We examined cell isolation, viability, and growth in adipose-derived stem cells harvested from whole adipose tissue subject to different cryopreservation lengths (2–1159 days from patients of varying ages (26–62 years. Subcutaneous abdominal adipose tissue was excised during abdominoplasties and was cryopreserved. The viability and number of adipose-derived stem cells isolated were measured after initial isolation and after 9, 18, and 28 days of growth. Data were analyzed with respect to cryopreservation duration and patient age. Significantly more viable cells were initially isolated from tissue cryopreserved 2 years, irrespective of patient age. However, this difference did not persist with continued growth and there were no significant differences in cell viability or growth at subsequent time points with respect to cryopreservation duration or patient age. Mesenchymal stem cell markers were maintained in all cohorts tested throughout the duration of the study. Consequently, longer cryopreservation negatively impacts initial live adipose-derived stem cell isolation; however, this effect is neutralized with continued cell growth. Patient age does not significantly impact stem cell isolation, viability, or growth. Cryopreservation of adipose tissue is an effective long-term banking method for isolation of adipose-derived stem cells in patients of varying ages.

Quality of raw cow milk in Republic of Macedonia determined through the testing of somatic cell count and total viable count

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Angelovski Ljupco

2008-11-01

Full Text Available Somatic cells count and total viable count are criteria used to estimate the compliance of raw cow milk with the Book of rules for demands for safety and hygiene and procedures for official controls of milk and milk products, Official Gazette of RM 157/2007. According to the given demands, raw milk operators are obliged to conduct all procedures and to guarantee that milk is in compliance with the criteria laid down in Book of rules. At the same time, Republic of Macedonia have to fulfill EU criteria laid down in Directive 92/46 (Council directive 92/46/EEC laying down the health rules for the production and placing on the market of raw milk, heat-treated milk and milkbased products for quality of raw milk as part of implementation of community legislation and milk production. The independent laboratory for milk quality control at FVM-Skopje, in frame of its activities in the period February- August 2008 has conducted a study for obtaining preliminary results for the situation with raw milk quality produced in R. of Macedonia for somatic cells counts and total viable count. In the study we analyzed 2065 samples for TVC and 1625 samples for SCC of raw milk samples produced in different parts of the country. From the tested samples only 41,8% fulfill criteria for SCC and 41,45% criteria for TVC lay down in Book of rules for 2008. Assessment of the results in light of Council Directive it is obvious that only 42,7% of the samples for SCC and 10,7% for TVC fulfill the criteria of Council Directive having in mind different requirements vs. Book of rules.

A comparison of the structure of ovules and seeds in Stemona (Stemonaceae) and Pentastemona (Pentastemonaceae)

NARCIS (Netherlands)

Bouman, F.; Devente, N.

1992-01-01

Stemona and Pentastemona differ clearly in the size and structure of their ovules and seeds. Especially the development, shape, and cell wall thickenings of the endotestal cells, and by consequence the origin of the seed ridges, show marked differences. The embryological and seed anatomical

Amniotic membrane seeded with mesenchymal adipose-derived stem cell for coverage of wound in third degree burn: An experimental study

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Mohammad Javad Fatemi

2014-09-01

Conclusion: Acellular amnion seeded with adipose-derived stem cell can result in faster wound healing and better histopathology characteristic. The amnion as a scaffold and the fat derived stem cells as healing accelerator are recommended for coverage of the 3rd degree burn wounds after excision and it may reduce the need for skin graft.

Radiation protective nursing intervene of 125I seed implantation in non-small cell lung carcinoma guided by CT

International Nuclear Information System (INIS)

Fu Li; Zhang Zuncheng; Yu Zhaochen; Zheng Guangjun; Tian Meirong

2009-01-01

Objective: To research radiation protective nursing intervene and important notice of 125 I seeds minimally invasive implantation in non-small cell lung carcinoma (NSCLC) by CT. Methods: Under the system of therapy planning system (TPS) and posologic validation, 125 I seeds were implanted in 89 cases of NSCLC patients. The consistent radiation protective nursing intervene was used in perioperative period management. The operative successful rate, therapeutic effect and complication rate, therapeutic effect and complication rate was observed. Results: The scientific radiation protective nursing intervene can ensure that the radioactive dose distribution of 125 I seed implantation brachytherapy is consistent with the principles of effective and minimally invasive. The operative successful rate was 100%. The local control rate and 1 year survival rate respectively was 97.4% and 92.2%. But the early and later incidence rate of radioactive damaging effect was 14.6% and 1.1% respectively. Leakage of radioactive contamination has not occurred. Conclusion: The consistent TPS and posologic validation 125 I seeds implantation integrated scientific radiation protective nursing intervene. It is very important to improve the therapeutic effect of NSCLC and reduce the incidence of complications. (authors)

A Combination of Histological, Physiological, and Proteomic Approaches Shed Light on Seed Desiccation Tolerance of the Basal Angiosperm Amborella trichopoda.

Science.gov (United States)

Villegente, Matthieu; Marmey, Philippe; Job, Claudette; Galland, Marc; Cueff, Gwendal; Godin, Béatrice; Rajjou, Loïc; Balliau, Thierry; Zivy, Michel; Fogliani, Bruno; Sarramegna-Burtet, Valérie; Job, Dominique

2017-07-28

Desiccation tolerance allows plant seeds to remain viable in a dry state for years and even centuries. To reveal potential evolutionary processes of this trait, we have conducted a shotgun proteomic analysis of isolated embryo and endosperm from mature seeds of Amborella trichopoda , an understory shrub endemic to New Caledonia that is considered to be the basal extant angiosperm. The present analysis led to the characterization of 415 and 69 proteins from the isolated embryo and endosperm tissues, respectively. The role of these proteins is discussed in terms of protein evolution and physiological properties of the rudimentary, underdeveloped, Amborella embryos, notably considering that the acquisition of desiccation tolerance corresponds to the final developmental stage of mature seeds possessing large embryos.

Primordial black holes as seeds of magnetic fields in the universe

Science.gov (United States)

Safarzadeh, Mohammadtaher

2018-06-01

Although it is assumed that magnetic fields in accretion disks are dragged from the interstellar medium, the idea is likely not applicable to primordial black holes (PBHs) formed in the early universe. Here we show that magnetic fields can be generated in initially unmagnetized accretion disks around PBHs through the Biermann battery mechanism, and therefore provide the small scale seeds of magnetic field in the universe. The radial temperature and vertical density profiles of these disks provide the necessary conditions for the battery to operate naturally. The generated seed fields have a toroidal structure with opposite sign in the upper and lower half of the disk. In the case of a thin accretion disk around a rotating PBH, the field generation rate increases with increasing PBH spin. At a fixed r/risco, where r is the radial distance from the PBH and risco is the radius of the innermost stable circular orbit, the battery scales as M-9/4, where M is the PBH's mass. The very weak dependency of the battery on accretion rate, makes this mechanism a viable candidate to provide seed fields in an initially unmagnetized accretion disk, following which the magnetorotational instability could take over.

Complementing endozoochorous seed dispersal patterns by donkeys and goats in a semi-natural island ecosystem.

Science.gov (United States)

Treitler, Julia Tabea; Drissen, Tim; Stadtmann, Robin; Zerbe, Stefan; Mantilla-Contreras, Jasmin

2017-12-19

Endozoochory is, in grazing systems, a substantial vector for seed dispersal. It can play an important role in vegetation dynamics, especially in colonization processes through seed input on the vegetation and on the soil seed bank. We investigated the endozoochorous seed input of donkeys and goats on a semi-natural island ecosystem in the Mediterranean. Through germination experiments, we assessed the viable seed content of the dung of these grazing animals to estimate their suitability and efficiency for seed dispersal of the vegetation types of the island. We show different dispersal patterns of donkeys and goats. Goats disperse a high number of diaspores from shrubs while donkeys disperse more diaspores of grasses. In addition, goats disperse plants of greater growth height and donkeys plants of shorter height. These dispersal patterns are in accordance with the vegetation types of which donkeys and goats disperse indicator species. Both, donkeys and goats, feed on and disperse species of the vegetation types, open grassland and temporarily wet grassland. In addition, goats feed on and disperse diagnostic species of the semi-open maquis and preforest formations. Overall, our results show that donkeys and goats are complementing each other in their endozoochorous seed dispersal potential. This emphasizes the importance of both grazing animals for the vegetation dynamics of the semi-natural island ecosystem. Therefore, the adaption of the goat management to a traditional land management based on directed transhumance might maintain and enrich vegetation types.

Identification of a seed coat-specific promoter fragment from the Arabidopsis MUCILAGE-MODIFIED4 gene.

Science.gov (United States)

Dean, Gillian H; Jin, Zhaoqing; Shi, Lin; Esfandiari, Elahe; McGee, Robert; Nabata, Kylie; Lee, Tiffany; Kunst, Ljerka; Western, Tamara L; Haughn, George W

2017-09-01

The Arabidopsis seed coat-specific promoter fragment described is an important tool for basic and applied research in Brassicaceae species. During differentiation, the epidermal cells of the Arabidopsis seed coat produce and secrete large quantities of mucilage. On hydration of mature seeds, this mucilage becomes easily accessible as it is extruded to form a tightly attached halo at the seed surface. Mucilage is composed mainly of pectin, and also contains the key cell wall components cellulose, hemicellulose, and proteins, making it a valuable model for studying numerous aspects of cell wall biology. Seed coat-specific promoters are an important tool that can be used to assess the effects of expressing biosynthetic enzymes and diverse cell wall-modifying proteins on mucilage structure and function. Additionally, they can be used for production of easily accessible recombinant proteins of commercial interest. The MUCILAGE-MODIFIED4 (MUM4) gene is expressed in a wide variety of plant tissues and is strongly up-regulated in the seed coat during mucilage synthesis, implying the presence of a seed coat-specific region in its promoter. Promoter deletion analysis facilitated isolation of a 308 base pair sequence (MUM4 0.3Pro ) that directs reporter gene expression in the seed coat cells of both Arabidopsis and Camelina sativa, and is regulated by the same transcription factor cascade as endogenous MUM4. Therefore, MUM4 0.3Pro is a promoter fragment that serves as a new tool for seed coat biology research.

Irradiation effect on the seed vigor, SOD activity and MDA content in germinating seeds of yellow-seeded and black-seeded rape seed (Brassica napus L.)

International Nuclear Information System (INIS)

Han Jixiang; Hu Danhong; Liu Houli

1993-01-01

Seeds of a set of near-isogenic lines (Brassica napus L.) with different seed coat color from yellow to black were irradiated by 60 Co γ-rays of 150 krad. Seed vigor, superoxide dismutase (SOD) and malondialdehyde (MDA) in germinating seeds were analysed. In these characters, no significant difference between yellow-seeded lines (YLs) and black-seeded lines (BLs) showed before irradiation. But after irradiation, SOD activity in YLs was lower than that in BLs. While MDA content in YLs was obviously higher that that in DLs. As a result of irradiation, seed vigor of YLs was lower than that in BLs. these results indicated that the irradiation resistance of rape seed was related to the level of SOD as well as protective structure or substances in seed coat and that the radiosensitivity of YLs was higher than that of DLs

Modulating and modeling aggregation of cell-seeded microcarriers in stirred culture system for macrotissue engineering.

Science.gov (United States)

Mei, Yang; Luo, Houyong; Tang, Qiang; Ye, Zhaoyang; Zhou, Yan; Tan, Wen-Song

2010-11-01

A recently developed protocol, "microtissue assembly" holds great promise to address the issue of limited mass transfer within engineered large tissue replacements (macrotissues), wherein small "building blocks" (microtissues) are prepared and then assembled into macrotissues. Previous studies suggested that aggregation behavior of microcarrier-based microtissues were very important for macrotissue engineering. However, a systematic study on the aggregation behavior of microtissues is still missing. In this study, to examine the aggregation behavior of microtissues, effects of key operation parameters in dynamic culture including cell seeding density, microcarrier concentration, L-ascorbic acid 2-phosphate (V(c)) and agitating speed were investigated. The aggregation process could be divided into three phases (i.e., lag, growth and stable). Aggregation efficiency (S) was found to be modulated by cell seeding density, microcarrier concentration, addition of V(c) and agitating speed. A mathematical model correlating the operation parameters with S at different phases of aggregation was developed and experimentally proved to be able to predict S with varied operation parameters. In the end, a cylindrical macrotissue (diameter × height: 2.0 cm × 0.8 cm) with fairly good integrity and cellularity and uniform cell distribution was successfully engineered through perfusion assembling microtissues with controlled S under selected culture conditions. Our study showed that aggregation of microtissues could be precisely modulated, which would definitely facilitate engineering macrotissues with high quality. Copyright © 2010 Elsevier B.V. All rights reserved.

Effect of 211At treating pollen and stigma on generative cells and seed setting of rice

International Nuclear Information System (INIS)

Jin Jiannan; Mo Shangwu; Liu Ning; Zhou Maolun; Zhang Shuyuan; Chen Fang; Zhang Yizheng; Gao Maoguo

1998-01-01

Low specific radioactivity (7.4 kBq/ml) 211 At treating pollen and stigma can obviously affect morphological structures and physiological functions of pollen, stigma and ovule or embryo sac cells, and cause injury. Results showed that because of the radiation effects the seed setting rate of rice was decreased, and the development of some embryos were affected and others became abnormal

Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry

Science.gov (United States)

Majeed, Muhammed; Majeed, Shaheen; Nagabhushanam, Kalyanam; Punnapuzha, Ardra; Philip, Sheena

2018-01-01

Accurate enumeration of bacterial count in probiotic formulation is imperative to ensure that the product adheres to regulatory standards and citation in consumer product label. Standard methods like plate count, can enumerate only replicating bacterial population under selected culture conditions. Viable but non culturable bacteria (VBNC) retain characteristics of living cells and can regain cultivability by a process known as resuscitation. This is a protective mechanism adapted by bacteria to evade stressful environmental conditions. B. coagulans MTCC 5856(LactoSpore®) is a probiotic endospore which can survive for decades in hostile environments without dividing. In the present study, we explored the use of flow cytometry to enumerate the viable count of B. coagulans MTCC 5856 under acidic and alkaline conditions, high temperature and in commercial formulations like compressed tablets and capsules. Flow cytometry (FCM) was comparable to plate count method when the spores were counted at physiological conditions. We show that VBNC state is induced in B. coagulans MTCC 5856by high temperature and acidic pH. The cells get resuscitated under physiological conditions and FCM was sensitive to detect the VBNC spores. Flow cytometry showed excellent ability to assess the viable spore count in commercial probiotic formulations of B. coagulans MTCC 5856. The results establish Flow cytometry as a reliable method to count viable bacteria in commercial probiotic preparations. Sporulation as well as existence as VBNC could contribute to the extreme stability of B. coagulans MTCC 5856. PMID:29474436

Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry.

Directory of Open Access Journals (Sweden)

Muhammed Majeed

Full Text Available Accurate enumeration of bacterial count in probiotic formulation is imperative to ensure that the product adheres to regulatory standards and citation in consumer product label. Standard methods like plate count, can enumerate only replicating bacterial population under selected culture conditions. Viable but non culturable bacteria (VBNC retain characteristics of living cells and can regain cultivability by a process known as resuscitation. This is a protective mechanism adapted by bacteria to evade stressful environmental conditions. B. coagulans MTCC 5856(LactoSpore® is a probiotic endospore which can survive for decades in hostile environments without dividing. In the present study, we explored the use of flow cytometry to enumerate the viable count of B. coagulans MTCC 5856 under acidic and alkaline conditions, high temperature and in commercial formulations like compressed tablets and capsules. Flow cytometry (FCM was comparable to plate count method when the spores were counted at physiological conditions. We show that VBNC state is induced in B. coagulans MTCC 5856by high temperature and acidic pH. The cells get resuscitated under physiological conditions and FCM was sensitive to detect the VBNC spores. Flow cytometry showed excellent ability to assess the viable spore count in commercial probiotic formulations of B. coagulans MTCC 5856. The results establish Flow cytometry as a reliable method to count viable bacteria in commercial probiotic preparations. Sporulation as well as existence as VBNC could contribute to the extreme stability of B. coagulans MTCC 5856.

Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry.

Science.gov (United States)

Majeed, Muhammed; Majeed, Shaheen; Nagabhushanam, Kalyanam; Punnapuzha, Ardra; Philip, Sheena; Mundkur, Lakshmi

2018-01-01

Accurate enumeration of bacterial count in probiotic formulation is imperative to ensure that the product adheres to regulatory standards and citation in consumer product label. Standard methods like plate count, can enumerate only replicating bacterial population under selected culture conditions. Viable but non culturable bacteria (VBNC) retain characteristics of living cells and can regain cultivability by a process known as resuscitation. This is a protective mechanism adapted by bacteria to evade stressful environmental conditions. B. coagulans MTCC 5856(LactoSpore®) is a probiotic endospore which can survive for decades in hostile environments without dividing. In the present study, we explored the use of flow cytometry to enumerate the viable count of B. coagulans MTCC 5856 under acidic and alkaline conditions, high temperature and in commercial formulations like compressed tablets and capsules. Flow cytometry (FCM) was comparable to plate count method when the spores were counted at physiological conditions. We show that VBNC state is induced in B. coagulans MTCC 5856by high temperature and acidic pH. The cells get resuscitated under physiological conditions and FCM was sensitive to detect the VBNC spores. Flow cytometry showed excellent ability to assess the viable spore count in commercial probiotic formulations of B. coagulans MTCC 5856. The results establish Flow cytometry as a reliable method to count viable bacteria in commercial probiotic preparations. Sporulation as well as existence as VBNC could contribute to the extreme stability of B. coagulans MTCC 5856.

Seed viability of five wild Saudi Arabian species by germination and X-ray tests.

Science.gov (United States)

Al-Hammad, B A; Al-Ammari, B S

2017-09-01

Our objective was to evaluate the usefulness of the germination vs. the X-ray test in determining the initial viability of seeds of five wild species ( Moringa peregrina , Abrus precatorius , Arthrocnemum macrostachyum , Acacia ehrenbergiana and Acacia tortilis ) from Saudi Arabia. Usually several days were required to determine the viability of all five species via germination tests. However, X-ray test will give immediate results on filled/viable seeds. Seeds of all species, except Acacia ehrenbergiana and Acacia tortilis showed high viability in both germination (96-72% at 25/15 °C, 94-70% at 35/25 °C) and X-ray (100-80%) test. Furthermore, there was a general agreement between the germination (19%, 14% at 25/15 °C and 17% and 12% at 35/25 °C) and X-ray (8%, 4%) tests in which seed viability of Acacia ehrenbergiana and Acacia tortilis was very low due to insect damaged embryo as shown in X-ray analysis. Seeds of Abruspreca torius have physical dormancy, which was broken by scarification in concentrated sulfuric acid (10 min), and they exhibited high viability in both the germination (83% at 25/15 °C and 81% at 35/25 °C) and X-ray (96%) tests. Most of the nongerminated seeds of the five species except those of Acacia ehrenbergiana and Acacia tortilis , were alive as judged by the tetrazolium test (TZ). Thus, for the five species examined, the X-ray test was proved to be a good and rapid predictor of seed viability.

Re-induction of desiccation tolerance after germination of Cedrela fissilis Vell. seeds.

Science.gov (United States)

Masetto, Tathiana E; Faria, Jose M; Fraiz, Ana C R

2014-09-01

This work aimed to characterize the re-induction of desiccation tolerance (DT) in germinated seeds, using polyethylene glycol (PEG 8000). Cell changes were investigated through cytological assays (cell viability and transmission electronic microscopy) as well as DNA integrity during loss and re-establishment of DT. The loss of DT was characterized by drying germinated seeds with different radicle lengths (1, 2, 3, 4 and 5 mm) in silica gel, decreasing the moisture content to ten percentage points intervals, followed by pre-humidification (100% RH / 24 h) and rehydration. To re-induce DT, germinated seeds were treated for 72 h with PEG (-2.04 MPa) and PEG (-2.04 MPa) + ABA (100 µM) before dehydration. Germinated seeds did not tolerate desiccation to 10% moisture content, irrespectively of the radicle length. However, when incubated in PEG, those with 1 and 2 mm long radicle attained 71% and 29% survival, respectively. The PEG+ABA treatment was efficient to re-establish DT in seeds with 1 mm long radicles (100% survival). The ultrastructural assays of the cells of germinated seeds with 2 and 5 mm length confirmed the obtained physiological results. Germinated seeds of C. fissilis constitute a useful tool for desiccation tolerance investigations.

Re-induction of desiccation tolerance after germination of Cedrela fissilis Vell. seeds

Directory of Open Access Journals (Sweden)

TATHIANA E. MASETTO

2014-09-01

Full Text Available This work aimed to characterize the re-induction of desiccation tolerance (DT in germinated seeds, using polyethylene glycol (PEG 8000. Cell changes were investigated through cytological assays (cell viability and transmission electronic microscopy as well as DNA integrity during loss and re-establishment of DT. The loss of DT was characterized by drying germinated seeds with different radicle lengths (1, 2, 3, 4 and 5 mm in silica gel, decreasing the moisture content to ten percentage points intervals, followed by pre-humidification (100% RH / 24 h and rehydration. To re-induce DT, germinated seeds were treated for 72 h with PEG (-2.04 MPa and PEG (-2.04 MPa + ABA (100 µM before dehydration. Germinated seeds did not tolerate desiccation to 10% moisture content, irrespectively of the radicle length. However, when incubated in PEG, those with 1 and 2 mm long radicle attained 71% and 29% survival, respectively. The PEG+ABA treatment was efficient to re-establish DT in seeds with 1 mm long radicles (100% survival. The ultrastructural assays of the cells of germinated seeds with 2 and 5 mm length confirmed the obtained physiological results. Germinated seeds of C. fissilis constitute a useful tool for desiccation tolerance investigations.

Soil seed banks Banco de sementes do solo

Directory of Open Access Journals (Sweden)

P.J. Christoffoleti

1998-01-01

Full Text Available The goal of this literature review is to discuss some of the major aspects of the soil seed bank, from its characteristics down to methodological aspects of its determination. Soil seed bank is the reservoir of viable seeds or of vegetative propagules that are present in the soil and that are able to recompose a natural vegetation. In the agroecossystems the soil seed bank is related to weeds, and the knowledge of its size and composition in terms of species can be used in the prediction of future infestations, to built simulation models of population establishment through time and also the definition of soil and cultural management programs, in order to have a rational use of herbicides.O objetivo desta revisão é discorrer sobre alguns pontos importantes no estudo do banco de sementes no solo, desde as sua características até aspectos metodológicos para sua determinação. Denomina-se de banco de sementes a reserva destas ou de propágulos vegetativos viáveis presentes no solo e que são capazes de recompor uma vegetação. Nos sistemas agrícolas o banco está relacionado com o estudo das plantas daninhas; o conhecimento do seu tamanho e das espécies que o compõem poderá ser utilizado na previsão de infestações futuras, na construção de modelos de estabelecimentos populacionais no tempo e consequentemente na definição de programas de manejo de solo e culturais, visando racionalizar a utilização de herbicidas.

Nutraceutical potential of hemp (Cannabis sativa L.) seeds and sprouts.

Science.gov (United States)

Frassinetti, Stefania; Moccia, Eleonora; Caltavuturo, Leonardo; Gabriele, Morena; Longo, Vincenzo; Bellani, Lorenza; Giorgi, Gianluca; Giorgetti, Lucia

2018-10-01

In this study the antioxidant effect of Cannabis sativa L. seeds and sprouts (3 and 5 days of germination) was evaluated. Total polyphenols, flavonoids and flavonols content, when expressed on dry weight basis, were highest in sprouts; ORAC and DPPH (in vitro assays), CAA-RBC (cellular antioxidant activity in red blood cells) and hemolysis test (ex vivo assays) evidenced a good antioxidant activity higher in sprouts than in seeds. Untargeted analysis by high resolution mass spectrometry in negative ion mode allowed the identification of main polyphenols (caffeoyltyramine, cannabisin A, B, C) in seeds and of ω-6 (linoleic acid) in sprouts. Antimutagenic effect of seeds and sprouts extracts evidenced a significant decrease of mutagenesis induced by hydrogen peroxide in Saccharomyces cerevisiae D7 strain. In conclusion our results show that C. sativa seeds and sprouts exert beneficial effects on yeast and human cells and should be further investigated as a potential functional food. Copyright © 2018. Published by Elsevier Ltd.

Structural Properties of Zinc Oxide Nanorods Grown on Al-Doped Zinc Oxide Seed Layer and Their Applications in Dye-Sensitized Solar Cells

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Kyung Ho Kim

2014-03-01

Full Text Available We fabricated zinc oxide (ZnO nanorods (NRs with Al-doped ZnO (AZO seed layers and dye-sensitized solar cells (DSSCs employed the ZnO NRs between a TiO2 photoelectrode and a fluorine-doped SnO2 (FTO electrode. The growth rate of the NRs was strongly dependent on the seed layer conditions, i.e., thickness, Al dopant and annealing temperature. Attaining a large particle size with a high crystallinity of the seed layer was vital to the well-aligned growth of the NRs. However, the growth was less related to the substrate material (glass and FTO coated glass. With optimized ZnO NRs, the DSSCs exhibited remarkably enhanced photovoltaic performance, because of the increase of dye absorption and fast carrier transfer, which, in turn, led to improved efficiency. The cell with the ZnO NRs grown on an AZO seed layer annealed at 350 °C showed a short-circuit current density (JSC of 12.56 mA/cm2, an open-circuit voltage (VOC of 0.70 V, a fill factor (FF of 0.59 and a power conversion efficiency (PCE, η of 5.20% under air mass 1.5 global (AM 1.5G illumination of 100 mW/cm2.

Detection of viable but non-culturable Escherichia coli O157:H7 by PCR in combination with propidium monoazide.

Science.gov (United States)

Zhong, Junliang; Zhao, Xihong

2018-01-01

The aim of this study was to evaluate the applicability of the conventional PCR detection method combined with propidium monoazide (PMA) treatment for the detection of viable but non-culturable (VBNC) state Escherichia coli O157:H7 in ground beef meatballs. Under low temperature, E. coli O157:H7 cells were induced into the VBNC state in ground beef meatballs at - 20 °C after 152 days. The optimal PMA concentration of 5 µg/mL was obtained in beef meatball samples, which could completely inhibit the DNA amplification on dead cells (10 6  cells/mL) but with no inhibition on viable cells. The established PMA-PCR assay revealed that the VBNC counts exceeded 10 7  CFU/mL in artificial contamination beef samples, which could be used for semi-quantitative detection of VBNC cells in beef meatball samples. This study indicated that the PMA-PCR assay might be a potential method for detection of VBNC state E . coli O157:H7 cells in food products.

Peroxiredoxin Expression of Human Osteosarcoma Cells Is Influenced by Cold Atmospheric Plasma Treatment.

Science.gov (United States)

Gümbel, Denis; Gelbrich, Nadine; Napp, Matthias; Daeschlein, Georg; Kramer, Axel; Sckell, Axel; Burchardt, Martin; Ekkernkamp, Axel; Stope, Matthias B

2017-03-01

To evaluate the potential involvement of redox-specific signalling pathways in cold atmospheric plasma (CAP)-induced apoptosis on human osteosarcoma cells. Osteosarcoma cell lines were treated with CAP with or without antioxidative agents and seeded in cell culture plates. Cell proliferation was determined by counting viable cells. Carrier gas-treated cells served as control. Peroxiredoxin (PRX) 1-3 expression and secretion were assessed. CAP treatment exhibited strongly attenuated proliferation rates. This effect was significantly attenuated by the addition of N-acetylcysteine (NAC). CAP-treated cells exhibited an increase of PRX 1 and 2 10 sec after treatment. The ratio of oxidized to reduced PRX1 and PRX2 was significantly altered with increasing cellular concentration of the oxidized dimer. Antioxidant supplementation with NAC increases proliferation of CAP-treated osteosarcoma cells, implicating an involvement of redox signalling. Activation of PRX1 and -2 indicate CAP affects redox homeostasis. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Schwann cell-mediated delivery of glial cell line-derived neurotrophic factor restores erectile function after cavernous nerve injury.

Science.gov (United States)

May, Florian; Buchner, Alexander; Schlenker, Boris; Gratzke, Christian; Arndt, Christian; Stief, Christian; Weidner, Norbert; Matiasek, Kaspar

2013-03-01

To evaluate the time-course of functional recovery after cavernous nerve injury using glial cell line-derived neurotrophic factor-transduced Schwann cell-seeded silicon tubes. Sections of the cavernous nerves were excised bilaterally (5 mm), followed by immediate bilateral surgical repair. A total of 20 study nerves per group were reconstructed by interposition of empty silicon tubes and silicon tubes seeded with either glial cell line-derived neurotrophic factor-overexpressing or green fluorescent protein-expressing Schwann cells. Control groups were either sham-operated or received bilateral nerve transection without nerve reconstruction. Erectile function was evaluated by relaparotomy, electrical nerve stimulation and intracavernous pressure recording after 2, 4, 6, 8 and 10 weeks. The animals underwent re-exploration only once, and were killed afterwards. The nerve grafts were investigated for the maturation state of regenerating nerve fibers and the fascular composition. Recovery of erectile function took at least 4 weeks in the current model. Glial cell line-derived neurotrophic factor-transduced Schwann cell grafts restored erectile function better than green fluorescent protein-transduced controls and unseeded conduits. Glial cell line-derived neurotrophic factor-transduced grafts promoted an intact erectile response (4/4) at 4, 6, 8 and 10 weeks that was overall significantly superior to negative controls (P cell line-derived neurotrophic factor-transduced grafts compared with negative controls (P = 0.018) and unseeded tubes (P = 0.034). Return of function was associated with the electron microscopic evidence of preganglionic myelinated nerve fibers and postganglionic unmyelinated axons. Schwann cell-mediated delivery of glial cell line-derived neurotrophic factor presents a viable approach for the treatment of erectile dysfunction after cavernous nerve injury. © 2013 The Japanese Urological Association.

Preparation and Low Temperature Short-term Storage for Synthetic Seeds of Caladium bicolor

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Mehpara MAQSOOD

2015-03-01

Full Text Available An efficient somatic embryo encapsulation and in vitro plant regeneration technique were established with Caladium bicolor, an important ornamental plant.Tuber derived embryogenic callus (95.50% was obtained on Murashige and Skoog (MS medium amended with 0.5 mg L-1 α-Naphthalene acetic acid (NAA + 0.5 mg L-1 6-Benzyladenine (BA. The embryogenic callus later differentiated into somatic embryos in the same plant growth regulators (PGRs added medium (NAA and BA. The induced embryos matured and developed into plantlets in NAA and BA added media; maximum plantlets development was observed at 1.0 mg L-1 NAA + 1.0 mg L-1 BA supplemented medium. Synthetic seeds were produced by encapsulating embryos in gel containing 3.0% sucrose + 3.0% sodium alginate and 100 mM of calcium chloride.The highest synthetic seed germination (97.6% was observed on medium supplemented with 1.0 mg L-1 NAA + 1.0 mg L-1 BA. The synthetic seeds were kept at low temperatures for storage; the encapsulated beads were viable and demonstrated good germination even after 12 weeks of storage at 4 °C. The plantlet recovery frequency was however declined with time. The synthetic seed derived plantlets were morphologically similar to the mother plant.

(Turcz) Schisandraceae seed extracts and cisplatin on cytotoxicity ...

African Journals Online (AJOL)

Purpose: Schisandra chinensis is a plant used in traditional Chinese and Russian medicine. An S. chinensis seed extract was tested for its ability to potentiate the effects of the anticancer agent cisplatin in MCF-7 breast cancer cells. Methods: S. chinensis seeds were extracted with ethanol and the ethanol was evaporated ...

Organic leek seed production - securing seed quality

DEFF Research Database (Denmark)

Deleuran, Lise Christina; Boelt, Birte

2011-01-01

To maintain integrity in organic farming, availability of organically produced GM-free seed of varieties adapted to organic production systems is of vital impor-tance. Despite recent achievements, organic seed supply for a number of vegetable species is insufficient. Still, in many countries...... seeds. Tunnel production is a means of securing seed of high genetic purity and quality, and organic leek (Allium porrum L.) seed production was tested in tunnels in Denmark. The present trial focused on steckling size and in all years large stecklings had a positive effect on both seed yield...

Organic Leek Seed Production - Securing Seed Quality

DEFF Research Database (Denmark)

Deleuran, L C; Boelt, B

2011-01-01

To maintain integrity in organic farming, availability of organically produced GM-free seed of varieties adapted to organic production systems is of vital impor-tance. Despite recent achievements, organic seed supply for a number of vegetable species is insufficient. Still, in many countries...... seeds. Tunnel production is a means of securing seed of high genetic purity and quality, and organic leek (Allium porrum L.) seed production was tested in tunnels in Denmark. The present trial focused on steckling size and in all years large stecklings had a positive effect on both seed yield...

Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.

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Bee Luan Khoo

Full Text Available Circulating tumor cells (CTCs are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation.Here, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56 (Breast cancer samples: 12-1275 CTCs/ml; Lung cancer samples: 10-1535 CTCs/ml rapidly from clinically relevant blood volumes (7.5 ml under 5 min. Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM, fluorescence in-situ hybridization (FISH (EML4-ALK or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA, and demonstrate concordance with the original tumor-biopsy samples.We have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS or proteomic analysis.

Viable Mycobacterium avium ssp. paratuberculosis isolated from calf milk replacer.

Science.gov (United States)

Grant, Irene R; Foddai, Antonio C G; Tarrant, James C; Kunkel, Brenna; Hartmann, Faye A; McGuirk, Sheila; Hansen, Chungyi; Talaat, Adel M; Collins, Michael T

2017-12-01

When advising farmers on how to control Johne's disease in an infected herd, one of the main recommendations is to avoid feeding waste milk to calves and instead feed calf milk replacer (CMR). This advice is based on the assumption that CMR is free of viable Mycobacterium avium ssp. paratuberculosis (MAP) cells, an assumption that has not previously been challenged. We tested commercial CMR products (n = 83) obtained from dairy farms around the United States by the peptide-mediated magnetic separation (PMS)-phage assay, PMS followed by liquid culture (PMS-culture), and direct IS900 quantitative PCR (qPCR). Conventional microbiological analyses for total mesophilic bacterial counts, coliforms, Salmonella, coagulase-negative staphylococci, streptococci, nonhemolytic Corynebacterium spp., and Bacillus spp. were also performed to assess the overall microbiological quality of the CMR. Twenty-six (31.3%) of the 83 CMR samples showed evidence of the presence of MAP. Seventeen (20.5%) tested positive for viable MAP by the PMS-phage assay, with plaque counts ranging from 6 to 1,212 pfu/50 mL of reconstituted CMR (average 248.5 pfu/50 mL). Twelve (14.5%) CMR samples tested positive for viable MAP by PMS-culture; isolates from all 12 of these samples were subsequently confirmed by whole-genome sequencing to be different cattle strains of MAP. Seven (8.4%) CMR samples tested positive for MAP DNA by IS900 qPCR. Four CMR samples tested positive by both PMS-based tests and 5 CMR samples tested positive by IS900 qPCR plus one or other of the PMS-based tests, but only one CMR sample tested positive by all 3 MAP detection tests applied. All conventional microbiology results were within current standards for whole milk powders. A significant association existed between higher total bacterial counts and presence of viable MAP indicated by either of the PMS-based assays. This represents the first published report of the isolation of viable MAP from CMR. Our findings raise concerns

Endothelial cell labeling with indium-111-oxine as a marker of cell attachment to bioprosthetic surfaces

International Nuclear Information System (INIS)

Sharefkin, J.B.; Lather, C.; Smith, M.; Rich, N.M.

1983-01-01

Canine vascular endothelium labeled with indium-111-oxine was used as a marker of cell attachment to vascular prosthetic surfaces with complex textures. Primarily cultured and freshly harvested endothelial cells both took up the label rapidly. An average of 72% of a 32 micro Ci labeling dose was taken up by 1.5 X 10(6) cells in 10 min in serum-free medium. Over 95% of freshly labeled cells were viable by trypan blue tests and only 5% of the label was released after 1 h incubations at 37 degrees C. Labeled and unlabeled cells had similar rates of attachment to plastic dishes. Scanning electron microscopic studies showed that labeled cells retained their ability to spread on tissue culture dishes even at low (1%) serum levels. Labeled endothelial cells seeded onto Dacron or expanded polytetrafluoroethylene vascular prostheses by methods used in current surgical models could be identified by autoradiography of microscopic sections of the prostheses, and the efficiency of cell attachment to the prosthesis could be measured by gamma counting. Indium-111 labeling affords a simple and rapid way to measure initial cell attachment to, and distribution on, vascular prosthetic materials. The method could also allow measurement of early cell loss from a flow surface in vivo by using external gamma imaging

Empty seeds are not always bad: simultaneous effect of seed emptiness and masting on animal seed predation.

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Ramón Perea

Full Text Available Seed masting and production of empty seeds have often been considered independently as different strategies to reduce seed predation by animals. Here, we integrate both phenomena within the whole assemblage of seed predators (both pre and post-dispersal and in two contrasting microsites (open vs. sheltered to improve our understanding of the factors controlling seed predation in a wind-dispersed tree (Ulmus laevis. In years with larger crop sizes more avian seed predators were attracted with an increase in the proportion of full seeds predated on the ground. However, for abundant crops, the presence of empty seeds decreased the proportion of full seeds predated. Empty seeds remained for a very long period in the tree, making location of full seeds more difficult for pre-dispersal predators and expanding the overall seed drop period at a very low cost (in dry biomass and allocation of C, N and P. Parthenocarpy (non-fertilized seeds was the main cause of seed emptiness whereas seed abortion was produced in low quantity. These aborted seeds fell prematurely and, thus, could not work as deceptive seeds. A proportion of 50% empty seeds significantly reduced ground seed predation by 26%. However, a high rate of parthenocarpy (beyond 50% empty seeds did not significantly reduce seed predation in comparison to 50% empty seeds. We also found a high variability and unpredictability in the production of empty seeds, both at tree and population level, making predator deception more effective. Open areas were especially important to facilitate seed survival since rodents (the main post-dispersal predators consumed seeds mostly under shrub cover. In elm trees parthenocarpy is a common event that might work as an adaptive strategy to reduce seed predation. Masting per se did not apparently reduce the overall proportion of seeds predated in this wind-dispersed tree, but kept great numbers of seeds unconsumed.

Iron and ferritin accumulate in separate cellular locations in Phaseolus seeds

DEFF Research Database (Denmark)

Cvitanich, Cristina; Przybylowicz, Wojciech J; Urbanski, Dorian Fabian

2010-01-01

and will assist in the production of staples with increased bioavailable iron. Results Here we reveal the distribution of iron in seeds of three Phaseolus species including thirteen genotypes of P. vulgaris, P. coccineus, and P. lunatus. We showed that high concentrations of iron accumulate in cells surrounding...... the provascular tissue of P. vulgaris and P. coccineus seeds. Using the Perls' Prussian blue method, we were able to detect iron in the cytoplasm of epidermal cells, cells near the epidermis, and cells surrounding the provascular tissue. In contrast, the protein ferritin that has been suggested as the major iron...... to P. vulgaris and P. coccineus, we did not observe iron accumulation in the cells surrounding the provascular tissues of P. lunatus cotyledons. A novel iron-rich genotype, NUA35, with a high concentration of iron both in the seed coat and cotyledons was bred from a cross between an Andean...

Activation of Arabidopsis seed hair development by cotton fiber-related genes.

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Xueying Guan

Full Text Available Each cotton fiber is a single-celled seed trichome or hair, and over 20,000 fibers may develop semi-synchronously on each seed. The molecular basis for seed hair development is unknown but is likely to share many similarities with leaf trichome development in Arabidopsis. Leaf trichome initiation in Arabidopsis thaliana is activated by GLABROUS1 (GL1 that is negatively regulated by TRIPTYCHON (TRY. Using laser capture microdissection and microarray analysis, we found that many putative MYB transcription factor and structural protein genes were differentially expressed in fiber and non-fiber tissues. Gossypium hirsutum MYB2 (GhMYB2, a putative GL1 homolog, and its downstream gene, GhRDL1, were highly expressed during fiber cell initiation. GhRDL1, a fiber-related gene with unknown function, was predominately localized around cell walls in stems, sepals, seed coats, and pollen grains. GFP:GhRDL1 and GhMYB2:YFP were co-localized in the nuclei of ectopic trichomes in siliques. Overexpressing GhRDL1 or GhMYB2 in A. thaliana Columbia-0 (Col-0 activated fiber-like hair production in 4-6% of seeds and had on obvious effects on trichome development in leaves or siliques. Co-overexpressing GhRDL1 and GhMYB2 in A. thaliana Col-0 plants increased hair formation in ∼8% of seeds. Overexpressing both GhRDL1 and GhMYB2 in A. thaliana Col-0 try mutant plants produced seed hair in ∼10% of seeds as well as dense trichomes inside and outside siliques, suggesting synergistic effects of GhRDL1 and GhMYB2 with try on development of trichomes inside and outside of siliques and seed hair in A. thaliana. These data suggest that a different combination of factors is required for the full development of trichomes (hairs in leaves, siliques, and seeds. A. thaliana can be developed as a model a system for discovering additional genes that control seed hair development in general and cotton fiber in particular.

El efecto de la disponibilidad de agua durante el crecimiento de Lycopersicon chilense sobre la capacidad de sus semillas para germinar a distintas temperaturas y concentraciones de manitol y NaCl Effect of water availability during the growth of Lycopersicon chilense on the capacity of their seeds to germinate at different temperatures and concentrations of manitol and NaCl

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CARLOS MALDONADO

2002-12-01

plantas de L. chilense producen semillas viables bajo las condiciones de déficit hídrico estudiadas. Sin embargo, la capacidad de germinar está limitada a un rango estrecho de condiciones ambientales. Se postulan algunas hipótesis para explicar su presencia en ambientes desérticos.The number of seeds produced by a plant can be limited by adverse environmental factors, which can also affect the seed development originating non-viable seeds. In the I Region, Chile we have observed that Lycopersicon chilense, a tomato species endemic to Atacama Desert, has high fruit and seed production, but a low plant recruitment. We hypothesize that this could be due to two factors: (a mother plants produce non-viable seeds because of the adverse conditions where they grow or (b seed are viable but the environmental conditions are not suitable for seed germination. We determined the germination capacity of L. chilense seeds, subjected to different temperatures and mannitol and NaCl concentration, from mother plants growing with plenty water supply (+H2O and other groups growing with water deficit (-H2O. Water supply affected the number of seeds germinated and the time needed by the seeds to germinate under different temperatures and water potential. Seeds from -H2O plants were able to germinate at lower water potentials and in shorter time than seeds from +H2O plants in the case of mannitol. The contrary occurred with NaCl, in which a higher seed germination and shorter time for germination occurred in seeds coming from +H2O plants. Seed germination of L chilense progressively decrease at water potentials lower than -0,5 MPa, being this decrease more evident with NaCl; in both cases there was a germination delay. The optimum temperature for seed germination was between 15 and 25 °C. Seeds of L. chilense did not germinate at temperatures lower than 8 °C or higher than 35 °C. Data show that although L. chilense plants produce viable seeds under the various water conditions studied