WorldWideScience

Sample records for viability cultures frozen

  1. The Effect of MSG (Monosodium Glutamate Addition on The Quality of Yoghurt Frozen Culture Starter Viewed Viability, pH Value and Acidity

    Directory of Open Access Journals (Sweden)

    Aris Sri Widati

    2012-02-01

    Full Text Available The objective of this study was to investigate wether the effect of percentage monosodium glutamat addition on the quality of yoghurt frozen culture starter viewed viability, pH value and acidity.The experimental design used in this study was Randomised Complete Design and the treatment were four levels of monosodium glutamate concentration respectively 0% (without monosodium glutamat 10%, 15% and 20% from medium. Each treatment were three times replicated. The research result showed that the difference of monosodium glutamate concentration  did not gave a significant effect (P>0.05 on viability of yoghurt frozen culture starter and acidity of yoghurt made by frozen culture starter but it gave a significant effect (P<0.05 on pH value. It can be concluded that different monosodium glutamate concentration had a different quality on frozen culture starter yoghurt. The addition of monosodium glutamate up to 20% necessarily indicate increase on quality of yoghurt frozen culture starter. Keywords: culture starter yoghurt, freezing, cryoprotectant

  2. The Effect of MSG (Monosodium Glutamate) Addition on The Quality of Yoghurt Frozen Culture Starter Viewed Viability, pH Value and Acidity

    OpenAIRE

    Aris Sri Widati; Abdul Manab; Teguh Hadi Waluyo

    2012-01-01

    The objective of this study was to investigate wether the effect of percentage monosodium glutamat addition on the quality of yoghurt frozen culture starter viewed viability, pH value and acidity.The experimental design used in this study was Randomised Complete Design and the treatment were four levels of monosodium glutamate concentration respectively 0% (without monosodium glutamat) 10%, 15% and 20% from medium. Each treatment were three times replicated. The research result showed that th...

  3. Non-invasive viability assessment of day-4 frozen-thawed human embryos using near infrared spectroscopy

    NARCIS (Netherlands)

    Vergouw, C.G.; Botros, L.L.; Judge, K.; Henson, M.; Roos, P.; Kostelijk, E.H.; Schats, R.; Twisk, J.W.; Hompes, P.G.A.; Sakkas, D.; Lambalk, C.B.

    2011-01-01

    This study investigated if metabolomic profiling of culture media using near infrared (NIR) spectroscopy was related to live-birth rates after single-embryo transfer of frozen-thawed embryos. Analysis of culture media of frozen-thawed embryos was performed by NIR spectroscopy. A viability score was

  4. A new bone banking technique to maintain osteoblast viability in frozen human iliac cancellous bone.

    Science.gov (United States)

    Oh, Jung-Hwan; Zöller, Joachim E; Kübler, Alexander

    2002-06-01

    The aim of this study was to develop a new cryopreservation technique to maintain the osteoblast viability in frozen iliac bone and to prove cell viability using cell culture techniques. Human iliac cancellous bones were frozen with and without 10% Me(2)SO at -80 degrees C. The tubes were kept in a -80 degrees C freezer for at least 2 days. After the storage period, the frozen bone was thawed by placing the tube in a 37 degrees C water bath. A serial enzymatic digestion technique using 0.2% collagenase was employed to isolate osteoblast-like cells from the bone. The cells that were released were inoculated into tissue culture flasks containing DMEM supplemented with 10% FCS. They were incubated at 37 degrees C in a humidified atmosphere of 95% air and 5% CO(2). Cells of the second passage were plated at a density of 5 x 10(3)cells/cm(2) in a 24-well plate and used for characterization. For characterization, WST-1 assay, determination of alkaline phosphatase, Type I collagen assay, osteocalcin assay, and von Kossa staining were used. The assays were performed at 3, 6, 9, and 12 days after plating the cells. Based on the results of this study, we conclude that the osteoblast-like cells in the frozen bone can survive, only when the bone is frozen with cryoprotectants to prevent injury during freezing and thawing.

  5. Frozen and fresh ovarian tissue require different culture media to promote in vitro development of bovine preantral follicles.

    Science.gov (United States)

    Castro, Simone Vieira; Carvalho, Adeline Andrade; Silva, Cleidson Manoel Gomes; Santos, Francielli Weber; Campello, Cláudio Cabral; de Figueiredo, José Ricardo; Rodrigues, Ana Paula Ribeiro

    2014-10-01

    The aim of this study was to evaluate the efficiency of different media in the in vitro culture of bovine preantral follicles that were used either fresh or following slow freezing treatment. Frozen and fresh noncultured or cultured ovarian fragments were processed for histological, viability, and cell proliferation analyses. For cryopreservation, a solution containing 1.5 M ethylene glycol was frozen in a programmable biological freezer. After thawing, a portion of the samples was destined for frozen controls. The remainder were cultured in vitro for 5 days in three media: α-MEM, McCoy, or M199. Samples from these culture media were collected on days 1 and 5 for quantification of reactive oxygen species (ROS) and for hormonal assays. In fresh-cultured tissues, the percentage of morphologically normal follicles was significantly higher when cultured in M199 compared to that in the other media. In frozen-cultured tissues, McCoy medium was significantly superior to the other media, and was the only treatment that helped in maintaining the viability similar to fresh and frozen controls. Upon quantification of the nucleolus organizer region, we observed greater proliferation of granulosa cells in the frozen-cultured tissues with McCoy medium, and lesser proliferation in fresh-cultured tissues only with α-MEM. In frozen-cultured tissues, ROS levels were highest at day 1 and progressively reduced during culture, independent of the media used. In conclusion, under the conditions used in this study, the M199 and McCoy media are recommended for the culture of follicles derived from fresh and frozen ovarian tissues, respectively.

  6. RELATIONSHIP BETWEEN MOTILITY AND VIABILITY PARAMETERS OF FROZEN-THAWED BULL SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    Eliška Špaleková

    2013-02-01

    Full Text Available The aim of this study was to determine relationship between parameters of spermatozoa motility (total motility - TM and progressive movement - PM and viability of bull frozen-thawed spermatozoa (dead spermatozoa ratio, apoptotic spermatozoa ratio and plasma membrane integrity. Motility parameters were evaluated using computer-assisted semen analysis (CASA. Parameters of spermatozoa viability were analysed using fluorescent dyes PNA-FITC (plasma membrane, Yo-Pro-1 and propidium iodide (PI. All bulls (n=6 were divided into two groups. First group (n=3 A – better bulls with total motility after thawing over 40% and the second group (n=3 B – with total motility lower than 40%. It was observed significantly (P<0.001 higher TM and PM in group A. No significant differences in velocity parameters and ALH between the group A and B were detected. Occurrence of spermatozoa with disrupted membranes, dead/necrotic spermatozoa and apoptotic spermatozoa was significantly lower in the group A. Bulls in the group A showed significantly higher cleavage rate of embryos. These motility and viability characteristics are associated with a higher embryo cleavage rate in in vitro fertilizatioThe aim of this study was to determine relationship between parameters of spermatozoa motility (total motility - TM and progressive movement - PM and viability of bull frozen-thawed spermatozoa (dead spermatozoa ratio, apoptotic spermatozoa ratio and plasma membrane integrity. Motility parameters were evaluated using computer-assisted semen analysis (CASA. Parameters of spermatozoa viability were analysed using fluorescent dyes PNA-FITC (plasma membrane, Yo-Pro-1 and propidium iodide (PI. All bulls (n=6 were divided into two groups. First group (n=3 A – better bulls with total motility after thawing over 40% and the second group (n=3 B – with total motility lower than 40%. It was observed significantly (P<0.001 higher TM and PM in group A. No significant differences in

  7. Abundance, viability and culturability of Antarctic bacteria

    Digital Repository Service at National Institute of Oceanography (India)

    LokaBharathi, P.A.; DeSouza, M.J.B.D.; Nair, S.; Chandramohan, D.

    The viability of total number of bacteria decide the mineralisation rate in any ecosystem and ultimately the fertility of the region. This study aims at establishing the extent of viability in the standing stock of the Antarctic bacterial population...

  8. Human ES cells: Starting Culture from Frozen Cells

    OpenAIRE

    Trish, Erin; Dimos, John; Eggan, Kevin

    2006-01-01

    Here we demonstrate how our lab begins a HuES human embryonic stem cell line culture from a frozen stock. First, a one to two day old ten cm plate of approximately one (to two) million irradiated mouse embryonic fibroblast feeder cells is rinsed with HuES media to remove residual serum and cell debris, and then HuES media added and left to equilibrate in the cell culture incubator. A frozen vial of cells from long term liquid nitrogen storage or a -80C freezer is sourced and quickly submer...

  9. Probiotic culture survival and implications in fermented frozen yogurt characteristics.

    Science.gov (United States)

    Davidson, R H; Duncan, S E; Hackney, C R; Eigel, W N; Boling, J W

    2000-04-01

    Low-fat ice cream mix was fermented with probiotic-supplemented and traditional starter culture systems and evaluated for culture survival, composition, and sensory characteristics of frozen product. Fermentations were stopped when the titratable acidity reached 0.15% greater than the initial titratable acidity (end point 1) or when the pH reached 5.6 (end point 2). Mix was frozen and stored for 11 wk at -20 degrees C. The traditional yogurt culture system contained the strains Streptococcus salivarius ssp. thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. The probiotic-supplemented system contained the traditional cultures as well as Bifidobacterium longum and Lactobacillus acidophilus. We compared recovery of Bifodobacterium by three methods, a repair-detection system with roll-tubes and plates on modified bifid glucose medium and plates with maltose + galactose reinforced clostridial medium. Culture bacteria in both systems did not decrease in the yogurt during frozen storage. The roll-tube method with modified bifid glucose agar and repair detection system provided at least one-half log10 cfu/ml higher recovery of B. longum compared with recoveries using modified bifid glucose agar or maltose + galactose reinforced clostridial agar on petri plates. No change in concentrations of lactose or protein for products fermented with either culture system occurred during storage. Acid flavor was more intense when product was fermented to pH 5.6, but yogurt flavor was not intensified. The presence of probiotic bacteria in the supplemented system seemed to cause no differences in protein and lactose concentration and sensory characteristics.

  10. Human ES cells: starting culture from frozen cells.

    Science.gov (United States)

    Trish, Erin; Dimos, John; Eggan, Kevin

    2006-11-09

    Here we demonstrate how our lab begins a HuES human embryonic stem cell line culture from a frozen stock. First, a one to two day old ten cm plate of approximately one (to two) million irradiated mouse embryonic fibroblast feeder cells is rinsed with HuES media to remove residual serum and cell debris, and then HuES media added and left to equilibrate in the cell culture incubator. A frozen vial of cells from long term liquid nitrogen storage or a -80 C freezer is sourced and quickly submerged in a 37 C water bath for quick thawing. Cells in freezing media are then removed from the vial and placed in a large volume of HuES media. The large volume of HuES media facilitates removal of excess serum and DMSO, which can cause HuES human embryonic stem cells to differentiate. Cells are gently spun out of suspension, and then re-suspended in a small volume of fresh HuES media that is then used to seed the MEF plate. It is considered important to seed the MEF plate by gently adding the HuES cells in a drop wise fashion to evenly disperse them throughout the plate. The newly established HuES culture plate is returned to the incubator for 48 hrs before media is replaced, then is fed every 24 hours thereafter.

  11. Viability of human corneal keratocytes during organ culture

    DEFF Research Database (Denmark)

    Møller-Pedersen, T; Møller, H J

    1996-01-01

    The viability of human corneal keratocytes was assessed during four weeks of 'closed system' organ culture at 31 degrees C. After 28 days of culturing, the entire keratocyte population was still alive and viable because all cells incorporated uridine; a parameter for RNA-synthesis. During the fir...... of keratan sulphate proteoglycan suggested that approximately 1% of the total content was lost during the period. In conclusion, our current organ culture technique can maintain a viable keratocyte population for four weeks; a viable stroma can be grafted within this period....

  12. Viability of Frozen-Thawed Epididymal Sperm of Garut Ram Cryopreserved with Modified Tris Extender

    Directory of Open Access Journals (Sweden)

    MUHAMMAD RIZAL

    2005-06-01

    Full Text Available Sperm collected from cauda epididymis is a source of male gametes. The purposes of this study was to evaluate an quality of frozen-thawed sperm of garut ram which was collected from cauda epididymis and cryopreserved with modified Tris extender, i.e: Tris extender (control, KT, Tris extender + 60 mM lactose (LS, and Tris extender + 60 mM lactose + 0.05% glutathione (GL. Quality of collected sperm including concentration, motility, live sperm, abnormality, cytoplasmic droplet, intact acrosomal cap (IAC, and intact plasma membrane (IPM were evaluated. Results showed that mean of sperm concentration, percentages of motility, live sperm, abnormality, cytoplasmic droplet, IAC, and IPM of fresh epididymal sperm were 13,993.33 million/ml, 70.83, 82.83, 10.83, 8.5, 85.83, and 81.33%, respectively. Sperm quality after equilibration for LS and GL were significantly (P<0.05 higher than KT. Mean percentages of post thawing sperm motility, live sperm, IAC, and IPM for GL (45, 54.5, 47.83, and 48.83% were significantly (P<0.05 higher than LS (40, 49.17, 43.83, and 44.5%, and KT (35, 42.5, 39.17, and 41.5%. Mean percentages of post thawing sperm motility, live sperm, IAC, and IPM for LS were significantly (P<0.05 higher than those of KT. Hence, frozen-thawed epididymal sperm of garut ram after slaughter and cryopreserved with Tris extender + 60 mM lactose (LS and Tris extender + 60 mM lactose + 0.05% glutathione (GL possibly can be used for artificial insemination (AI or in vitro embryo production program.

  13. Wastewater treatment to enhance the economic viability of microalgae culture.

    Science.gov (United States)

    Pires, J C M; Alvim-Ferraz, M C M; Martins, F G; Simões, M

    2013-08-01

    Microalgae culture is still not economically viable and it presents some negative environmental impacts, concerning water, nutrient and energy requirements. In this context, this study aims to review the recent advances on microalgal cultures in wastewaters to enhance their economic viability. We focused on three different culture concepts: (1) suspended cell systems, (2) cell immobilization, and (3) microalgae consortia. Cultures with suspended cells are the most studied. The nutrient removal efficiencies are usually high for wastewaters of different sources. However, biomass harvesting is difficult and a costly process due to the small cell size and lower culture density. On the other hand, the cell immobilization systems showed to be the solution for this problem, having as main limitation the nutrient diffusion from bulk to cells, which results in a reduced nutrient removal efficiency. The consortium between microalgae and bacteria enhances the growth of both microorganisms. This culture concept showed to be a promising technology to improve wastewater treatment, regarding not only nutrient removal but also biomass harvesting by bioflocculation. The aggregation mechanism must be studied in depth to find the process parameters that would lead to an effective and cheap harvesting process.

  14. Sperm preparation after freezing improves motile sperm count, motility, and viability in frozen-thawed sperm compared with sperm preparation before freezing-thawing process.

    Science.gov (United States)

    Palomar Rios, A; Gascón, A; Martínez, J V; Balasch, S; Molina Botella, I

    2017-10-09

    The aim of this study is to evaluate which cryopreservation protocol, freezing before or after swim-up, optimizes cryopreservation outcomes in terms of motile sperm count, motility, morphology, and viability, and also to establish whether sperm viability could be assessed based on sperm motility. Fifty-three fresh and 53 swim-up prepared samples were considered for the first experiment. In parallel, total motility evaluation by CASA system (computer-assisted sperm analyzer) and hypoosmotic swelling test (HOS-test) was performed in each sample to compare the viability results of both methods. In the second experiment, 21 normozoospermic semen samples and 20 semen samples from male factor patients were included. After fresh ejaculate evaluation, the semen sample of each patient was divided into two aliquots, one of them was frozen before swim-up and the other was frozen after swim-up. Motility, sperm count, morphology, and viability were evaluated after thawing. A linear regression model allows prediction of HOS-test viability results based on total motility: HOS = 1.38 + 0.97 · TM (R (2) = 99.10, residual mean squares = 9.51). Freezing before sperm selection leads to higher total and progressive motility, total motile sperm count, and viability rates than when sperm selection is performed before freezing (P < 0.005 in all cases). In fact, sperm selection prior to freezing reaches critical values when subfertile patients are considered. To conclude, total motility evaluation can predict HOS-test viability results, resulting in a more objective and less time-consuming method to assess viability. In addition, sperm freezing prior to swim-up selection must be considered in order to achieve better outcomes after thawing, especially in patients presenting poor sperm baseline.

  15. Evaluation of different cryoprotective agents in maintenance of viability of Helicobacter pylori in stock culture media

    Directory of Open Access Journals (Sweden)

    Daryoush Davoudi Oskouei

    2010-12-01

    Full Text Available Four different cryoprotective supplemented stock media were evaluated for maintaining better survival and recovery of H. pylori type strain NCTC 11637 at two different maintenance temperatures of -20°C and -80°C after one month preservation as frozen stocks. The spread plate colony count method was used to investigate the recovery rate of H. pylori from equally inoculated bacterial suspensions in differently prepared stock cultures. After the preservation of H. pylori for one month in different cryoprotectant-supplemented stock media, the recovery rates for -20°C obtained for stock cultures supplemented with dimethyl sulfoxide (DMSO, polyethylene glycol (PEG, glycerol and glycerol+sucrose, as well as controls with and without human serum alone were 7.13, 6.97, 7.93, 7.99, 6.95 and 0.0 log CFU/ml, respectively. Maintenance of bacteria at -80°C gave statistically higher recovery rates compared to preservation at -20°C with the values of 8.55, 8.24, 8.59, 8.66, 8.01 and 0.0 log CFU/ml for these above mentioned stock cultures. The stock cultures supplemented with glycerol+sucrose and glycerol showed the highest recovery rates, 7.99 and 7.93 for -20°C vs. 8.66 and 8.59 for -80°C respectively, which were statistically different from the others. Our study revealed that H. pylori type strain NCTC 11637 could be better preserved at -80°C than -20°C. The best stock media which supported viability or culturability of bacteria were brain heart infusion broth (BHI+glycerol+human serum and BHI+glycerol+sucrose+human serum, where the latter yielded the higher recovery rate.

  16. Viability of dielectrophoretically trapped neuronal cortical cells in culture

    NARCIS (Netherlands)

    Heida, Tjitske; Vulto, P; Rutten, Wim; Marani, Enrico

    2001-01-01

    Negative dielectrophoretic trapping of neural cells is an efficient way to position neural cells on the electrode sites of planar micro-electrode arrays. The preservation of viability of the neural cells is essential for this approach. This study investigates the viability of postnatal cortical rat

  17. Non-invasive metabolomic profiling of embryo culture media and morphology grading to predict implantation outcome in frozen-thawed embryo transfer cycles.

    Science.gov (United States)

    Li, Xiong; Xu, Yan; Fu, Jing; Zhang, Wen-Bi; Liu, Su-Ying; Sun, Xiao-Xi

    2015-11-01

    Assessment of embryo viability is a crucial component of in vitro fertilization and currently relies largely on embryo morphology and cleavage rate. Because morphological assessment remains highly subjective, it can be unreliable in predicting embryo viability. This study investigated the metabolomic profiling of embryo culture media using near-infrared (NIR) spectroscopy for predicting the implantation potential of human embryos in frozen-thawed embryo transfer (FET) cycles. Spent embryo culture media was collected on day 4 after thawed embryo transfer (n = 621) and analysed using NIR spectroscopy. Viability scores were calculated using a predictive multivariate algorithm of fresh embryos with known pregnancy outcomes. The mean viability indices of embryos resulting in clinical pregnancy following FET were significantly higher than those of non-implanted embryos and differed between the 0, 50, and 100 % implantation groups. Notably, the 0 % group index was significantly lower than the 100 % implantation group index (-0.787 ± 0.382 vs. 1.064 ± 0.331, P  0.05). NIR metabolomic profiling of thawed embryo culture media is independent of morphology and correlates with embryo implantation potential in FET cycles. The viability score alone or in conjunction with morphologic grading is a more objective marker for implantation outcome in FET cycles than morphology alone.

  18. Viability after thawing and dilution of simultaneously cryopreserved vaccinal Babesia bovis and Babesia bigemina strains cultured in vitro.

    Science.gov (United States)

    Mangold, A J; Vanzini, V R; Echaide, I E; de Echaide, S T; Volpogni, M M; Guglielmone, A A

    1996-02-01

    A live, frozen experimental vaccine containing Babesia bovis and Babesia bigemina multiplied in vitro was stored in liquid nitrogen after simultaneous cryopreservation using glycerol as cryoprotectant. The viability of the vaccine was tested by inoculating (subcutaneously) three groups of seven steers each, 2, 12 and 24 h after thawing at 40 degrees C and dilution to obtain a dose of 2 x 10(7) of each organism. All vaccinated cattle developed detectable parasitaemia in thin and/or thick blood smears. No statistically significant differences in the prepatent period were detected amongst the cattle groups (analysis of variance). This prepatent period was 12.3 days for B. bovis and 8.4 days for B. bigemina. Vaccinal organisms derived from in vitro culture systems may replace antigens obtained from in vivo culture to produce vaccine against cattle babesiosis.

  19. Equine ovarian tissue viability after cryopreservation and in vitro culture

    Science.gov (United States)

    The efficiency of several cryoprotective agents were compared using both slow-freezing and vitrification methods. Results indicate that the viability of ovarian tissue cells increases when DMSO (slow-freezing) and ethylene glycol (vitrification) are used....

  20. Conservation science in a terrorist age: the impact of airport security screening on the viability and DNA integrity of frozen felid spermatozoa.

    Science.gov (United States)

    Gloor, Kayleen T; Winget, Doug; Swanson, William F

    2006-09-01

    In response to growing terrorism concerns, the Transportation Security Administration now requires that all checked baggage at U.S. airports be scanned through a cabinet x-ray system, which may increase risk of radiation damage to transported biologic samples and other sensitive genetic material. The objective of this study was to investigate the effect of these new airport security regulations on the viability and DNA integrity of frozen felid spermatozoa. Semen was collected from two domestic cats (Felis silvestris catus) and one fishing cat (Prionailurus viverrinus), cryopreserved in plastic freezing straws, and transferred into liquid nitrogen dry shippers for security screening. Treatment groups included frozen samples from each male scanned once or three times using a Transportation Security Administration-operated cabinet x-ray system, in addition to non-scanned samples (i.e., negative control) and samples previously scanned three times and exposed to five additional high-intensity x-ray bursts (i.e., positive control). Dosimeters placed in empty dry shippers were used to quantify radiation exposure. Following treatment, straws were thawed and spermatozoa analyzed for post-thaw motility (percentage motile and rate of progressive movement), acrosome status, and DNA integrity using single-cell gel electrophoresis (i.e., the comet assay). Dosimeter measurements determined that each airport screening procedure produced approximately 16 mrem of radiation exposure. Our results indicated that all levels of radiation exposure adversely affected (P 0.05) among treatment groups. Results also showed that the amount of double-stranded DNA damage was greater (P < 0.05) in sperm samples from both cat species scanned three times compared to samples scanned once or negative controls. Findings suggest that new airport security measures may cause radiation-induced damage to frozen spermatozoa and other valuable biologic samples transported on passenger aircraft and that

  1. Viabilidade de células de levedura em massas congeladas de pão francês Leavening cells viability in frozen French bread dough

    Directory of Open Access Journals (Sweden)

    Carolina Pereira Kechinski

    2010-05-01

    Full Text Available A elaboração e o armazenamento de pães crus congelados têm se mostrado vantajosos para os pontos de venda e para o consumidor final. O primeiro, pela redução da perda e menor exigência de espaço físico e mão-de-obra especializada, e o segundo, por disponibilizar pães sempre frescos. O presente trabalho objetivou verificar a influência do armazenamento sobre a viabilidade das células de levedura em massas congeladas de pão francês. Para tanto, as massas foram elaboradas seguindo a metodologia de massa direta: os ingredientes (pré-mistura para pão francês, fermento fresco e água foram misturados até o desenvolvimento completo do glúten; então foi dividida em porções de 60g e modelada. O congelamento foi realizado em ultracongelador a -32°C, e o armazenamento foi feito em câmaras com temperaturas de -18°C e -40°C. Para a estimativa de vida de prateleira, analisou-se a cinética da redução da viabilidade das leveduras. Esse teste foi realizado em placas de Petri contendo meio Sabouraud acrescido de cloranfenicol 0,2% a 28°C durante cinco dias. A cinética de redução das células de levedura seguiu uma reação de primeira ordem, cujas constantes de velocidades foram 0,027 dias-1 (R²=0,931 e 0,026 dias-¹ (R²=0,938 para as temperaturas de -18°C e -40°C, respectivamente. Para ambas as temperaturas, a redução em 50% da população de leveduras se deu em aproximadamente 25 dias.The preparation and storage of frozen bread has proved to be advantageous for the market and to the final consumer. First because of loss reduction and need for smaller physical storage space and workforce, second because it provides always fresh bread. This study aimed to investigate the influence of storage on the viability of yeast cells in frozen French bread. In order to that, frozen French breads were prepared by the direct-dough method: the ingredients (premix for French bread, fresh yeast and water were mixed until the dough was

  2. Non-destructive monitoring of viability in an ex vivo organ culture model of osteochondral tissue.

    Science.gov (United States)

    Elson, K M; Fox, N; Tipper, J L; Kirkham, J; Hall, R M; Fisher, J; Ingham, E

    2015-06-30

    Organ culture is an increasingly important tool in research, with advantages over monolayer cell culture due to the inherent natural environment of tissues. Successful organ cultures must retain cell viability. The aim of this study was to produce viable and non-viable osteochondral organ cultures, to assess the accumulation of soluble markers in the conditioned medium for predicting tissue viability. Porcine femoral osteochondral plugs were cultured for 20 days, with the addition of Triton X-100 on day 6 (to induce necrosis), camptothecin (to induce apoptosis) or no toxic additives. Tissue viability was assessed by the tissue destructive XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxyanilide tetrazolium salt) assay method and LIVE/DEAD® staining of the cartilage at days 0, 6 and 20. Tissue structure was assessed by histological evaluation using haematoxylin & eosin and safranin O. Conditioned medium was assessed every 3-4 days for glucose depletion, and levels of lactate dehydrogenase (LDH), alkaline phosphatase (AP), glycosaminoglycans (GAGs), and matrix metalloproteinase (MMP)-2 and MMP-9. Necrotic cultures immediately showed a reduction in glucose consumption, and an immediate increase in LDH, GAG, MMP-2 and MMP-9 levels. Apoptotic cultures showed a delayed reduction in glucose consumption and delayed increase in LDH, a small rise in MMP-2 and MMP-9, but no significant effect on GAGs released into the conditioned medium. The data showed that tissue viability could be monitored by assessing the conditioned medium for the aforementioned markers, negating the need for tissue destructive assays. Physiologically relevant whole- or part-joint organ culture models, necessary for research and pre-clinical assessment of therapies, could be monitored this way, reducing the need to sacrifice tissues to determine viability, and hence reducing the sample numbers necessary.

  3. Long-term viability and differentiation of bovine oviductal monolayers: bidimensional versus three-dimensional culture.

    Science.gov (United States)

    Gualtieri, R; Mollo, V; Braun, S; Barbato, V; Fiorentino, I; Talevi, R

    2012-10-15

    Different in vitro models have been developed to study the interaction of gametes and embryos with the maternal tract. In cattle, the interaction of the oviduct with gametes and embryos have been classically studied using oviductal explants or monolayers (OMs). Explants are well differentiated but have to be used within 24 h after collection, whereas OMs can be used for a longer time after cell confluence but dedifferentiate during culture, losing cell polarity and ciliation. Herein, OMs were cultured either in M199 plus 10% fetal calf serum or in a semidefined culture medium (Gray's medium), in an immersed condition on collagen-coated coated microporous polyester or polycarbonate inserts under air-liquid interface conditions. The influence of culture conditions on long-term viability and differentiation of OMs was evaluated through scanning electron microscopy, localization of centrin and tubulin at the confocal laser scanning microscope, and assessment of maintenance of viability of sperm bound to OMs. Findings demonstrated that OMs cultured in an immersed condition with Gray's medium retain a better morphology, do not exhibit signs of crisis at least until 3 wks postconfluence, and maintain the viability of bound sperm significantly better than parallel OMs cultured in M199 plus 10% fetal calf serum. OM culture with Gray's medium in air-liquid interface conditions on porous inserts promotes cell polarity, ciliation, and maintenance of bound sperm viability at least until 3 wks postconfluence. In conclusion, oviduct culture in Gray's medium in an immersed or air-liquid condition allows long-term culture and, in the latter case, also ciliation of bovine OMs, and may represent in vitro systems that mimick more closely the biological processes modulated by the oviduct in vivo. Copyright © 2012 Elsevier Inc. All rights reserved.

  4. Comparative studies of cellular viability levels on 2D and 3D in vitro culture matrices.

    Science.gov (United States)

    Gargotti, M; Lopez-Gonzalez, U; Byrne, H J; Casey, A

    2017-09-18

    In this study, the cellular viability and function of immortalized human cervical and dermal cells are monitored and compared in conventional 2D and two commercial 3D membranes, Collagen and Geltrex, of varying working concentration and volume. Viability was monitored with the aid of the Alamar Blue assay, cellular morphology was monitored with confocal microscopy, and cell cycle studies and cell death mechanism studies were performed with flow cytometry. The viability studies showed apparent differences between the 2D and 3D culture systems, the differences attributed in part to the physical transition from 2D to 3D environment causing alterations to effective resazurin concentration, uptake and conversion rates, which was dependent on exposure time, but also due to the effect of the membrane itself on cellular function. These effects were verified by flow cytometry, in which no significant differences in viable cell numbers between 2D and 3D systems were observed after 24 h culture. The results showed the observed effect was different after shorter exposure periods, was also dependent on working concentration of the 3D system and could be mediated by altering the culture vessel size. Cell cycle analysis revealed cellular function could be altered by growth on the 3D substrates and the alterations were noted to be dependent on 3D membrane concentration. The use of 3D culture matrices has been widely interpreted to result in "improved viability levels" or "reduced" toxicity or cellular "resistance" compared to cells cultured on traditional 2D systems. The results of this study show that cellular health and viability levels are not altered by culture in 3D environments, but their normal cycle can be altered as indicated in the cell cycle studies performed and such variations must be accounted for in studies employing 3D membranes for in vitro cellular screening.

  5. Viability and physiological state transitions of Rhizopus oligosporus sporangiospores in tempe starter culture

    NARCIS (Netherlands)

    Thanh, N.V.; Rombouts, F.M.; Nout, M.J.R.

    2007-01-01

    The viability and various physiological characteristics of individual sporangiospores of Rhizopus oligosporus in tempe starter cultures that had been stored for 8, 10, 16 and 30 months were examined by flow cytometry in combination with fluorescent dyes. Besides live, dead, and dormant spores we

  6. Towards an understanding of the influence of national culture on organisational viability: An exploratory study

    Directory of Open Access Journals (Sweden)

    Awuzie Bankole O.

    2017-06-01

    Full Text Available ABSTRACT Viability connotes a system’s ability to become ultra-stable through effective self-regulation of its internal processes and information processing among its subsystems. Applying this to an infrastructure delivery system (IDS context, this study proposes that an IDS can successfully deliver on client requirements only if they attain and maintain viability. Research into the influence of National Culture (NC on an IDS’s viability appears to be lacking; hence this study. Adopting a multi-case study, qualitative research design, this study explores three IDSs involved in the delivery of infrastructure projects in two different NC contexts; Nigeria and the United Kingdom. 25 semi-structured interviews were conducted across the cases to provide for an in-depth understanding of existing interactions between participants in these delivery systems: client/project sponsor; main contractor and sub-contractors and to understand the influence of the prevailing national culture on such interactions, if any. Findings indicate that NC in project delivery environments influence the ability of IDSs to attain viability, especially as it pertains to the sustenance of Team Quality Attributes (TWQ within the system. Based on these findings, it is expected that in modelling IDSs for viability, adequate consideration should be given to the prevailing NC by project managers and planners.

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

    Science.gov (United States)

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

    2017-01-01

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

  8. A simple colony-formation assay in liquid medium, termed 'tadpoling', provides a sensitive measure of Saccharomyces cerevisiae culture viability.

    Science.gov (United States)

    Welch, Aaron Z; Koshland, Douglas E

    2013-12-01

    Here we describe the first high-throughput amenable method of quantifying Saccharomyces cerevisiae culture viability. Current high-throughput methods of assessing yeast cell viability, such as flow cytometry and SGA analysis, do not measure the percentage viability of a culture but instead measure cell vitality or colony fitness, respectively. We developed a method, called tadpoling, to quantify the percentage viability of a yeast culture, with the ability to detect as few as one viable cell amongst ~10(8) dead cells. The most important feature of this assay is the exploitation of yeast colony formation in liquid medium. Utilizing a microtiter dish, we are able to observe a range of viability of 100% to 0.0001%. Comparison of tadpoling to the traditional plating method to measure yeast culture viability reveals that, for the majority of Saccharomyces species analyzed there is no significant difference between the two methods. In comparison to flow cytometry using propidium iodide, the high-throughput method of measuring yeast culture viability, tadpoling is much more accurate at culture viabilities < 1%. Thus, we show that tadpoling provides an easy, inexpensive, space-saving method, amenable to high-throughput screens, for accurately measuring yeast cell viability. Copyright © 2013 John Wiley & Sons, Ltd.

  9. The influence of micronutrients in cell culture: a reflection on viability and genomic stability.

    Science.gov (United States)

    Arigony, Ana Lúcia Vargas; de Oliveira, Iuri Marques; Machado, Miriana; Bordin, Diana Lilian; Bergter, Lothar; Prá, Daniel; Henriques, João Antonio Pêgas

    2013-01-01

    Micronutrients, including minerals and vitamins, are indispensable to DNA metabolic pathways and thus are as important for life as macronutrients. Without the proper nutrients, genomic instability compromises homeostasis, leading to chronic diseases and certain types of cancer. Cell-culture media try to mimic the in vivo environment, providing in vitro models used to infer cells' responses to different stimuli. This review summarizes and discusses studies of cell-culture supplementation with micronutrients that can increase cell viability and genomic stability, with a particular focus on previous in vitro experiments. In these studies, the cell-culture media include certain vitamins and minerals at concentrations not equal to the physiological levels. In many common culture media, the sole source of micronutrients is fetal bovine serum (FBS), which contributes to only 5-10% of the media composition. Minimal attention has been dedicated to FBS composition, micronutrients in cell cultures as a whole, or the influence of micronutrients on the viability and genetics of cultured cells. Further studies better evaluating micronutrients' roles at a molecular level and influence on the genomic stability of cells are still needed.

  10. The Influence of Micronutrients in Cell Culture: A Reflection on Viability and Genomic Stability

    Science.gov (United States)

    Arigony, Ana Lúcia Vargas; de Oliveira, Iuri Marques; Bordin, Diana Lilian; Prá, Daniel; Pêgas Henriques, João Antonio

    2013-01-01

    Micronutrients, including minerals and vitamins, are indispensable to DNA metabolic pathways and thus are as important for life as macronutrients. Without the proper nutrients, genomic instability compromises homeostasis, leading to chronic diseases and certain types of cancer. Cell-culture media try to mimic the in vivo environment, providing in vitro models used to infer cells' responses to different stimuli. This review summarizes and discusses studies of cell-culture supplementation with micronutrients that can increase cell viability and genomic stability, with a particular focus on previous in vitro experiments. In these studies, the cell-culture media include certain vitamins and minerals at concentrations not equal to the physiological levels. In many common culture media, the sole source of micronutrients is fetal bovine serum (FBS), which contributes to only 5–10% of the media composition. Minimal attention has been dedicated to FBS composition, micronutrients in cell cultures as a whole, or the influence of micronutrients on the viability and genetics of cultured cells. Further studies better evaluating micronutrients' roles at a molecular level and influence on the genomic stability of cells are still needed. PMID:23781504

  11. The Influence of Micronutrients in Cell Culture: A Reflection on Viability and Genomic Stability

    Directory of Open Access Journals (Sweden)

    Ana Lúcia Vargas Arigony

    2013-01-01

    Full Text Available Micronutrients, including minerals and vitamins, are indispensable to DNA metabolic pathways and thus are as important for life as macronutrients. Without the proper nutrients, genomic instability compromises homeostasis, leading to chronic diseases and certain types of cancer. Cell-culture media try to mimic the in vivo environment, providing in vitro models used to infer cells' responses to different stimuli. This review summarizes and discusses studies of cell-culture supplementation with micronutrients that can increase cell viability and genomic stability, with a particular focus on previous in vitro experiments. In these studies, the cell-culture media include certain vitamins and minerals at concentrations not equal to the physiological levels. In many common culture media, the sole source of micronutrients is fetal bovine serum (FBS, which contributes to only 5–10% of the media composition. Minimal attention has been dedicated to FBS composition, micronutrients in cell cultures as a whole, or the influence of micronutrients on the viability and genetics of cultured cells. Further studies better evaluating micronutrients' roles at a molecular level and influence on the genomic stability of cells are still needed.

  12. Viability of 'Candidatus Liberibacter asiaticus' prolonged by addition of citrus juice to culture medium.

    Science.gov (United States)

    Parker, Jennifer K; Wisotsky, Sarah R; Johnson, Evan G; Hijaz, Faraj M; Killiny, Nabil; Hilf, Mark E; De La Fuente, Leonardo

    2014-01-01

    Huanglongbing, or citrus greening disease, is associated with infection by the phloem-limited bacterium 'Candidatus Liberibacter asiaticus'. Infection with 'Ca. L. asiaticus' is incurable; therefore, knowledge regarding 'Ca. L. asiaticus' biology and pathogenesis is essential to develop a treatment. However, 'Ca. L. asiaticus' cannot currently be successfully cultured, limiting its study. To gain insight into the conditions conducive for growth of 'Ca. L. asiaticus' in vitro, 'Ca. L. asiaticus' inoculum obtained from seed of fruit from infected pomelo trees (Citrus maxima 'Mato Buntan') was added to different media, and cell viability was monitored for up to 2 months using quantitative polymerase chain reaction in conjunction with ethidium monoazide. Media tested included one-third King's B (K), K with 50% juice from the infected fruit, K with 50% commercially available grapefruit juice, and 100% commercially available grapefruit juice. Results show that juice-containing media dramatically prolong viability compared with K in experiments reproduced during 2 years using different juice sources. Furthermore, biofilm formed at the air-liquid interface of juice cultures contained 'Ca. L. asiaticus' cells, though next-generation sequencing indicated that other bacterial genera were predominant. Chemical characterization of the media was conducted to discuss possible factors sustaining 'Ca. L. asiaticus' viability in vitro, which will contribute to future development of a culture medium for 'Ca. L. asiaticus'.

  13. Effect of Selenate on Viability and Selenomethionine Accumulation of Chlorella sorokiniana Grown in Batch Culture

    Science.gov (United States)

    Vílchez, Carlos; Torronteras, Rafael; Vigara, Javier; Gómez-Jacinto, Veronica; Janzer, Nora; Gómez-Ariza, José-Luis; Márová, Ivana

    2014-01-01

    The aim of this work was to study the effect of Se(+VI) on viability, cell morphology, and selenomethionine accumulation of the green alga Chlorella sorokiniana grown in batch cultures. Culture exposed to sublethal Se concentrations of 40 mg·L−1 (212 μM) decreased growth rates for about 25% compared to control. A selenate EC50 value of 45 mg·L−1 (238.2 μM) was determined. Results showed that chlorophyll and carotenoids contents were not affected by Se exposure, while oxygen evolution decreased by half. Ultrastructural studies revealed granular stroma, fingerprint-like appearance of thylakoids which did not compromise cell activity. Unlike control cultures, SDS PAGE electrophoresis of crude extracts from selenate-exposed cell cultures revealed appearance of a protein band identified as 53 kDa Rubisco large subunit of Chlorella sorokiniana, suggesting that selenate affects expression of the corresponding chloroplast gene as this subunit is encoded in the chloroplast DNA. Results revealed that the microalga was able to accumulate up to 140 mg·kg−1 of SeMet in 120 h of cultivation. This paper shows that Chlorella sorokiniana biomass can be enriched in the high value aminoacid SeMet in batch cultures, while keeping photochemical viability and carbon dioxide fixation activity intact, if exposed to suitable sublethal concentrations of Se. PMID:24688385

  14. Effect of Selenate on Viability and Selenomethionine Accumulation of Chlorella sorokiniana Grown in Batch Culture

    Directory of Open Access Journals (Sweden)

    Živan Gojkovic

    2014-01-01

    Full Text Available The aim of this work was to study the effect of Se(+VI on viability, cell morphology, and selenomethionine accumulation of the green alga Chlorella sorokiniana grown in batch cultures. Culture exposed to sublethal Se concentrations of 40 mg·L−1 (212 μM decreased growth rates for about 25% compared to control. A selenate EC50 value of 45 mg·L−1 (238.2 μM was determined. Results showed that chlorophyll and carotenoids contents were not affected by Se exposure, while oxygen evolution decreased by half. Ultrastructural studies revealed granular stroma, fingerprint-like appearance of thylakoids which did not compromise cell activity. Unlike control cultures, SDS PAGE electrophoresis of crude extracts from selenate-exposed cell cultures revealed appearance of a protein band identified as 53 kDa Rubisco large subunit of Chlorella sorokiniana, suggesting that selenate affects expression of the corresponding chloroplast gene as this subunit is encoded in the chloroplast DNA. Results revealed that the microalga was able to accumulate up to 140 mg·kg−1 of SeMet in 120 h of cultivation. This paper shows that Chlorella sorokiniana biomass can be enriched in the high value aminoacid SeMet in batch cultures, while keeping photochemical viability and carbon dioxide fixation activity intact, if exposed to suitable sublethal concentrations of Se.

  15. Viability Assessment of Genipa americana L. (Rubiaceae Embryonic Axes after Cryopreservation Using In Vitro Culture

    Directory of Open Access Journals (Sweden)

    Izulmé Rita Imaculada Santos

    2016-01-01

    Full Text Available Embryonic axes excised from seeds of Genipa americana L. desiccated to different water contents were successfully cryopreserved by rapidly plunging seed samples directly into liquid nitrogen. Control and cryopreserved embryonic axes were excised and grown in WPM culture medium for viability assessment. All control embryonic axes (−LN2 excised from fully hydrated seeds (43.89% moisture content germinated after 21 days of culture in vitro. These high germination percentages persisted even after the water content of the seeds was as low as 6.79%. After freezing in liquid nitrogen high germination percentages, 93%, 96%, and 93%, were observed for embryonic axes excised from seeds dehydrated to 13.26%, 9.57%, and 6.79 moisture content, respectively. The cryopreservation technique described here is recommended for long term conservation of G. americana germplasm.

  16. Effects of Trypsinization on Viability of Equine Chondrocytes in Cell Culture

    Directory of Open Access Journals (Sweden)

    B. C. Sutradhar, J. Park, G. Hong, S. H. Choi and G. Kim*

    2010-10-01

    Full Text Available Trypsin is an essential reagent for routine cell culture work. In the cultivation of mammalian cells, it has been extensively used for cell isolation from tissues or cell dislodging in subculturing. It may damage the cell membrane in contact of cells during long trypsinization. However, there is no specific report on time-dependent effect of trypsinization on cells. In the present study, we investigated the time dependent effects of trypsinization on equine chondrocytes. Cell viability after trypsinization with 0.25% trypsin-EDTA for 5 to 60 minutes was quantified by trypan blue exclusion assay, propidium iodide-Hoechst double staining, flow cytometry analysis and XTT assay. The results showed that trypsin-EDTA decreased the proliferation of equine chondrocytes depending on the exposure time of trypsinization. After 20 and 60 minutes of trypsinization, the cell membranes were strongly affected and the percentages of viable cells reduced to 91% and 85% respectively detected by trypan blue exclusion assay. Similar results were observed both in flow cytometric evaluation and propidium iodide-Hoechst double staining. The XTT assay result also showed decreased cell viability with the extended time of trypsinization. In order to minimize the time dependant cytotoxicity of trypsinization, as minimum as short time exposure is suggestive that maximizes live cell isolation from tissue as well as subculture of equine chondrocytes or other cells.

  17. Detached leaf culture: viability to evaluate 2,4-D toxicity symptoms in cotton apex leaves.

    Science.gov (United States)

    da C Centurion, Maria Aparecida P; Correia, Núbia M; Sanches, Vivaldo César R

    2005-01-01

    To study the viability of detached leaf culture technique, studies were carried out with detached leaves from cotton apex (true trilobed leaves). The prepared leaves were sprayed with 2,4-D amine and ester, at rates of 10, 30, 70, and 100% of the recommended doses. Detached leaves without herbicide spray were used as controls. Simultaneously, a greenhouse experiment was conducted with the same treatments as used for the detached leaves experiment. Toxicity was measured through a 0-to-5 grading according to the percentage of affected leaf area in the detached leaves experiment or examining the affected rate of whole plant as indicated in the greenhouse. Results showed that the ester form of the herbicide induced earlier and more severe toxicity symptoms in detached leaves and greenhouse grown plants. Positive and significant correlations (p cotton plants grown in the greenhouse (r= - 0.92 and -0.92, respectively).

  18. Effect of diluent type on viability of yeasts enumerated from foods or pure culture.

    Science.gov (United States)

    Mian, M A; Fleet, G H; Hocking, A D

    1997-04-01

    The effects of seven diluent types on the viability of yeasts enumerated from foods and in pure culture were studied. The diluents were laboratory glass distilled water; saline water (0.85% NaCl), sodium phosphate buffer (0.1 M, pH 7.0), 0.1% peptone, 0.1% yeast extract, 0.1% peptone in 0.1 M sodium phosphate buffer, pH 7.0, and 0.1% malt extract. For all foods studied, dilution in 0.1% peptone gave the highest counts, with saline and phosphate buffer diluents giving lower counts than those obtained with distilled water. When seven species of yeast were enumerated in pure culture, highest counts were obtained using 0.1% peptone as the diluent and, with three exceptions, all species gave higher counts when diluted in diluents other than distilled water. When yeast suspensions were held in diluents for up to 2 h before plating, cell death occurred. The extent of death was highest in distilled water, saline and phosphate buffer diluents. Cell death also occurred in 0.1% peptone, yeast extract and malt extract, but to a lesser degree.

  19. Redefining the effect of salt on thermophilic starter cell viability, culturability and metabolic activity in cheese.

    Science.gov (United States)

    Hickey, C D; Fallico, V; Wilkinson, M G; Sheehan, J J

    2018-02-01

    This study investigated the differential effect of salt concentration in the outside and inside layers of brine salted cheeses on viability, culturability and enzyme activity of starter bacteria. The high-salt environment of the outside layer caused a sharp decrease in L. helveticus viability as measured by traditional plate counts. Remarkably, this was associated with lower release of intracellular enzymes (LDH), reduced levels of proteolysis and larger membrane integrity as measured by flow cytometry (FC) following classical Live/Dead staining. FC analysis of light scattering properties highlighted a significant reduction in size and granularity of the microbiota located in the cheese surface, suggestive of cell shrinkage and condensation of internal macromolecules probably due to hyperosmotic stress. The microbiota of the cheese surface were found to experience greater oxidative stress, as measured by FC analysis of the total levels of reactive oxygen species, compared to that of the interior layer. These results lead us to postulate that the physiology and health status of the microbiota were significantly different in the outer and inner layers of the cheese. The hyperosmotic environment of the outer layer resulted in reduced cell lysis, as measurable by assays based upon membrane integrity, but rather triggered cell death via mechanisms involving cell shrinkage and ROS-mediated damage of vital intracellular components. This study challenges the current thinking on how salt controls microbial activity in ripening cheese, especially in cheeses which are brine salted as local variations in biochemical ripening indices can differ significantly from the outside to the inside of a ripening cheese. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Disruption of Functional Activity of Mitochondria during MTT Assay of Viability of Cultured Neurons.

    Science.gov (United States)

    Surin, A M; Sharipov, R R; Krasil'nikova, I A; Boyarkin, D P; Lisina, O Yu; Gorbacheva, L R; Avetisyan, A V; Pinelis, V G

    2017-06-01

    The MTT assay based on the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium in the cell cytoplasm to a strongly light absorbing formazan is among the most commonly used methods for determination of cell viability and activity of NAD-dependent oxidoreductases. In the present study, the effects of MTT (0.1 mg/ml) on mitochondrial potential (ΔΨm), intracellular NADH, and respiration of cultured rat cerebellum neurons and isolated rat liver mitochondria were investigated. MTT caused rapid quenching of NADH autofluorescence, fluorescence of MitoTracker Green (MTG) and ΔΨm-sensitive probes Rh123 (rhodamine 123) and TMRM (tetramethylrhodamine methyl ester). The Rh123 signal, unlike that of NADH, MTG, and TMRM, increased in the nucleoplasm after 5-10 min, and this was accompanied by the formation of opaque aggregates of formazan in the cytoplasm and neurites. Increase in the Rh123 signal indicated diffusion of the probe from mitochondria to cytosol and nucleus due to ΔΨm decrease. Inhibition of complex I of the respiratory chain decreased the rate of formazan formation, while inhibition of complex IV increased it. Inhibition of complex III and ATP-synthase affected only insignificantly the rate of formazan formation. Inhibition of glycolysis by 2-deoxy-D-glucose blocked the MTT reduction, whereas pyruvate increased the rate of formazan formation in a concentration-dependent manner. MTT reduced the rate of oxygen consumption by cultured neurons to the value observed when respiratory chain complexes I and III were simultaneously blocked, and it suppressed respiration of isolated mitochondria if substrates oxidized by NAD-dependent dehydrogenases were used. These results demonstrate that formazan formation in cultured rat cerebellum neurons occurs primarily in mitochondria. The initial rate of formazan formation may serve as an indicator of complex I activity and pyruvate transport rate.

  1. Simulated Microgravity and 3D Culture Enhance Induction, Viability, Proliferation and Differentiation of Cardiac Progenitors from Human Pluripotent Stem Cells

    Science.gov (United States)

    Jha, Rajneesh; Wu, Qingling; Singh, Monalisa; Preininger, Marcela K.; Han, Pengcheng; Ding, Gouliang; Cho, Hee Cheol; Jo, Hanjoong; Maher, Kevin O.; Wagner, Mary B.; Xu, Chunhui

    2016-01-01

    Efficient generation of cardiomyocytes from human pluripotent stem cells is critical for their regenerative applications. Microgravity and 3D culture can profoundly modulate cell proliferation and survival. Here, we engineered microscale progenitor cardiac spheres from human pluripotent stem cells and exposed the spheres to simulated microgravity using a random positioning machine for 3 days during their differentiation to cardiomyocytes. This process resulted in the production of highly enriched cardiomyocytes (99% purity) with high viability (90%) and expected functional properties, with a 1.5 to 4-fold higher yield of cardiomyocytes from each undifferentiated stem cell as compared with 3D-standard gravity culture. Increased induction, proliferation and viability of cardiac progenitors as well as up-regulation of genes associated with proliferation and survival at the early stage of differentiation were observed in the 3D culture under simulated microgravity. Therefore, a combination of 3D culture and simulated microgravity can be used to efficiently generate highly enriched cardiomyocytes. PMID:27492371

  2. Using simulation to study difficult clinical issues: prenatal counseling at the threshold of viability across american and dutch cultures

    NARCIS (Netherlands)

    Geurtzen, R.; Hogeveen, M.; Rajani, A.K.; Chitkara, R.; Antonius, T.A.; Heijst, A.F. van; Draaisma, J.M.; Halamek, L.P.

    2014-01-01

    OBJECTIVE: Prenatal counseling at the threshold of viability is a challenging yet critically important activity, and care guidelines differ across cultures. Studying how this task is performed in the actual clinical environment is extremely difficult. In this pilot study, we used simulation as a

  3. Evaluation of viability and proliferative activity of human urothelial cells cultured onto xenogenic tissue-engineered extracellular matrices.

    LENUS (Irish Health Repository)

    Davis, Niall F

    2011-04-01

    To evaluate the viability and proliferative activity of human urothelial cells (HUCs) cultured on tissue-engineered extracellular matrix scaffolds and to assess the potential of extracellular matrixes to support the growth of HUCs in their expected in vivo urine environment.

  4. Enrichment of probiotic ice cream with different dietary fibers: Structural characteristics and culture viability.

    Science.gov (United States)

    Akalın, A S; Kesenkas, H; Dinkci, N; Unal, G; Ozer, E; Kınık, O

    2018-01-01

    This study evaluated the effect of 5 dietary fibers (apple, orange, oat, bamboo, and wheat) on the physicochemical, rheological, and textural characteristics; sensory properties; and culture viability of probiotic ice cream stored at -18°C for 180 d. The presence of orange and apple fibers increased the titratable acidity, decreased the lightness (color) value of the ice creams, and enhanced the red and yellow coloration. Compared with the control sample, the consistency indices and apparent viscosities of the experimental samples increased with the addition of all dietary fibers except oat fiber. The highest viscosity was obtained in the sample fortified with apple fiber, whereas the ice cream containing orange fiber showed the highest hardness after d 60 of storage. The addition of orange and apple fibers significantly increased melting resistance; however, panelists did not generally like these samples in terms of taste-flavor. All ice creams had viable counts of Lactobacillus acidophilus of ≥7 log cfu/g during storage except the samples with orange and bamboo fiber. Bifidobacterium lactis counts were also found to be >6 log cfu/g in those samples until d 150 of storage. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  5. High-content assays for characterizing the viability and morphology of 3D cancer spheroid cultures.

    Science.gov (United States)

    Sirenko, Oksana; Mitlo, Trisha; Hesley, Jayne; Luke, Steve; Owens, Windsor; Cromwell, Evan F

    2015-09-01

    There is an increasing interest in using three-dimensional (3D) spheroids for modeling cancer and tissue biology to accelerate translation research. Development of higher throughput assays to quantify phenotypic changes in spheroids is an active area of investigation. The goal of this study was to develop higher throughput high-content imaging and analysis methods to characterize phenotypic changes in human cancer spheroids in response to compound treatment. We optimized spheroid cell culture protocols using low adhesion U-bottom 96- and 384-well plates for three common cancer cell lines and improved the workflow with a one-step staining procedure that reduces assay time and minimizes variability. We streamlined imaging acquisition by using a maximum projection algorithm that combines cellular information from multiple slices through a 3D object into a single image, enabling efficient comparison of different spheroid phenotypes. A custom image analysis method was implemented to provide multiparametric characterization of single-cell and spheroid phenotypes. We report a number of readouts, including quantification of marker-specific cell numbers, measurement of cell viability and apoptosis, and characterization of spheroid size and shape. Assay performance was assessed using established anticancer cytostatic and cytotoxic drugs. We demonstrated concentration-response effects for different readouts and measured IC50 values, comparing 3D spheroid results to two-dimensional cell cultures. Finally, a library of 119 approved anticancer drugs was screened across a wide range of concentrations using HCT116 colon cancer spheroids. The proposed methods can increase performance and throughput of high-content assays for compound screening and evaluation of anticancer drugs with 3D cell models.

  6. Effect of culture age, protectants, and initial cell concentration on viability of freeze-dried cells of Metschnikowia pulcherrima.

    Science.gov (United States)

    Spadaro, Davide; Ciavorella, Annalisa Alessandra; Lopez-Reyes, Jorge Giovanny; Garibaldi, Angelo; Gullino, Maria Lodovica

    2010-10-01

    The effect of freeze-drying using different lyoprotectants at different concentrations on the viability and biocontrol efficacy of Metschnikowia pulcherrima was evaluated. The effects of initial yeast cell concentration and culture age on viability were also considered. Yeast cells grown for 36 h were more resistant to freeze-drying than were 48 h cells. An initial concentration of 10⁸ cells·mL⁻¹ favoured the highest survival after freeze-drying. When maltose (25%, m/v) was used as protectant, a high cell viability was obtained (64.2%). Cells maintained a high viability after 6 months of storage at 4 °C. The biocontrol efficacy of freeze-dried cells was similar to the activity of fresh cells on 'Gala' apples and was slightly lower on 'Golden Delicious' apples. After optimizing freeze-drying conditions, the viability of M. pulcherrima cells was similar to that obtained in other studies. The results constitute a first step towards the commercial development of M. pulcherrima as a biocontrol agent.

  7. Effect of freezing and thawing rates on the post-thaw viability of boar spermatozoa frozen in FlatPacks and Maxi-straws.

    Science.gov (United States)

    Eriksson, B M; Rodriguez-Martinez, H

    2000-11-01

    The effects of different freezing and thawing rates on the post-thaw motility and membrane integrity of boar spermatozoa, processed as split samples in Maxi-straws or flat PET-plastic packages (FlatPack) were studied. A programmable freezing device was used to obtain freezing rates of either 20, 50 or 80 degrees C/min. Thawing of the samples was performed in a bath of circulating water; for 40s at 50 degrees C or 27s at 70 degrees C for Maxi-straws and 23s at 35 degrees C, 13s at 50 degrees C or 8s at 70 degrees C for the FlatPacks. Sperm motility was assessed both visually and with a computer assisted semen analysis (CASA) apparatus, while plasma membrane integrity was assessed using the fluorescent probes Calcein AM and ethidium homodimer-1. Temperature changes during freezing and thawing were monitored in both forms of packaging. Values for motile spermatozoa, sperm velocity and lateral head displacement variables were significantly (pstraws, with superior results at higher thawing rates. Freezing at 50 degrees C/min yielded better motility than 20 or 80 degrees C/min, although the effect was rather small. Neither freezing rate nor thawing rate had any effect on membrane integrity (p>0.05). A significant boar effect was seen for several parameters. The most striking difference in temperature courses between containers was a 4-5-fold lowering of the thawing rate, between -20 and 0 degrees C, in the center of the Maxi-straw, compared with the FlatPack. This is apparently due to the insulating effect of the thawed water in the periphery of the Maxi-straw. The improvement in sperm motility seen when using the FlatPack appears to be related to the rapid thawing throughout the sample, which decreases the risk of cell damage due to recrystallization during thawing. Since sperm motility patterns have been reported to be correlated with fertility both in vitro and in vivo it is speculated that the use of the FlatPack might improve the results when using frozen

  8. Effect of starter culture and inulin addition on microbial viability, texture, and chemical characteristics of whole or skim milk Kefir

    Directory of Open Access Journals (Sweden)

    Flávia Daiana Montanuci

    2012-12-01

    Full Text Available The effect of inulin addition and starters (Kefir grains or commercial starter culture on the microbial viability, texture, and chemical characteristics of Kefir beverages prepared with whole or skim milk was evaluated during refrigerated storage. The type of starter did not influence microbial viability during the storage of the beverages, but the chemical and textural changes (decreases in pH, lactose concentration, and inulin and increased acidity, firmness, and syneresis were more pronounced in the formulations fermented with grains than those fermented with the starter culture. The addition of inulin did not influence acidity or viability of lactic acid bacteria, but in general, its effect on the survival of acetic acid bacteria, Lactococcus and yeasts, firmness, and syneresis depended on the type of milk and starter culture used. Generally, the yeast, acetic acid bacteria, and Leuconostoc counts increased or remained unchanged, while the total population of lactic acid bacteria and Lactococcus were either reduced by 1 to 2 logs or remained unchanged during storage.

  9. Increased viability but decreased culturability of Mycobacterium avium subsp. paratuberculosis in macrophages from inflammatory bowel disease patients under Infliximab treatment.

    Science.gov (United States)

    Nazareth, Nair; Magro, Fernando; Appelberg, Rui; Silva, Jani; Gracio, Daniela; Coelho, Rosa; Cabral, José Miguel; Abreu, Candida; Macedo, Guilherme; Bull, Tim J; Sarmento, Amélia

    2015-12-01

    Mycobacterium avium subsp. paratuberculosis (MAP) has long been implicated as a triggering agent in Crohn's disease (CD). In this study, we investigated the growth/persistence of both M. avium subsp. hominissuis (MAH) and MAP, in macrophages from healthy controls (HC), CD and ulcerative colitis patients. For viability assessment, both CFU counts and a pre16SrRNA RNA/DNA ratio assay (for MAP) were used. Phagolysosome fusion was evaluated by immunofluorescence, through analysis of LAMP-1 colocalization with MAP. IBD macrophages were more permissive to MAP survival than HC macrophages (a finding not evident with MAH), but did not support MAP active growth. The lower MAP CFU counts in macrophage cultures associated with Infliximab treatment were not due to increased killing, but possibly to elevation in the proportion of intracellular dormant non-culturable MAP forms, as MAP showed higher viability in those macrophages. Increased MAP viability was not related to lack of phagolysosome maturation. The predominant induction of MAP dormant forms by Infliximab treatment may explain the lack of MAP reactivation during anti-TNF therapy of CD but does not exclude the possibility of MAP recrudescence after termination of therapy.

  10. Frozen Shoulder

    Science.gov (United States)

    ... surgery is one measure prescribed to prevent frozen shoulder. Symptoms Pain from frozen shoulder is usually dull or aching. It is typically ... disease and when you move your arm. The pain is usually located over the outer ... er discussing your symptoms and medical history, your doctor will examine your ...

  11. CELL VIABILITY IN X-RAY IRRADIATED HUMAN LENS EPITHELIAL CELL (HLEC CULTURE BY USING TRYPAN BLUE EXCLUSION TEST

    Directory of Open Access Journals (Sweden)

    Rajesh

    2015-12-01

    Full Text Available INTRODUCTION Cataract is one of major cause of blindness, 44% cases of blindness is due to cataract only. In developing countries such as India cataract is more common and it manifest earlier in life than the most developed countries of the world. Patients receiving therapeutic radiation may have developed cataracts as well as radiation retinopathy and optic nerve damage. In this study we are aimed to compare effects of X-ray irradiation on cell viability of HLEC (Human lens epithelial cell culture by using trypan blue dye exclusion test against the unexposed control HLEC culture. Total 20 cataract patients from Dept. of Ophthalmology of Lata Mangeshkar Hospital, Nagpur were included in this study. The capsulorhexis sample from cataract surgery taken for study from eye OT. The capsulorhexis sample without any x-ray exposure taken as control. For culture of HLEC’s RPMI 1640 medium containing 10% fetal calf serum used. The HLEC cultures were X-ray irradiated with increasing count of skull limit parameters of range 150kV, 250mAs and 50mA for 5sec using GE® 650 M.A. X-ray machine. The Trypan blue dye exclusion test was used to analyze cell viability. The slides were analyzed using Olympus® BX51 Research microscope with oil immersion and 100 were observed per sample. The cell viability was calculated using Epi info® (version 6.0, as subject means with standard deviation. Statistical significance will be defined at P≤0.001. Out of 100 cases the 12 were female and 8 were male. Mean age of both male and female patients were 62.45±3.18(SD. A positive linear correlation was found (r:0.819, p<0.001 between the No. of X-Ray exposure and death cells per 100 observed LECs.

  12. Viability of Baylisascaris procyonis Eggs

    OpenAIRE

    Shira C Shafir; Sorvillo, Frank J.; Sorvillo, Teresa; Eberhard, Mark L.

    2011-01-01

    Infection with Baylisascaris procyonis roundworms is rare but often fatal and typically affects children. We attempted to determine parameters of viability and methods of inactivating the eggs of these roundworms. Loss of viability resulted when eggs were heated to 62°C or desiccated for 7 months but not when frozen at –15°C for 6 months.

  13. Viability and contribution to proteolysis of an adjunct culture of Lactobacillus plantarum in two model cheese systems: Cheddar cheese‐type and soft‐cheese type

    National Research Council Canada - National Science Library

    Milesi, M.M; McSweeney, P.L.H; Hynes, E.R

    2008-01-01

    Aims:  The influence of the cheese‐making process, ripening conditions and primary starter on the viability and proteolytic activity of an adjunct culture of Lactobacillus plantarum I91 was assessed in two miniature cheese models...

  14. Hydrochloric acid alters the effect of L-glutamic acid on cell viability in human neuroblastoma cell cultures.

    Science.gov (United States)

    Croce, Nicoletta; Bernardini, Sergio; Di Cecca, Stefano; Caltagirone, Carlo; Angelucci, Francesco

    2013-07-15

    l-Glutamic acid (l-glutamate) is used to induce excitotoxicity and test neuroprotective compounds in cell cultures. However, because l-glutamate powder is nearly insoluble in water, many manufacturers recommend reconstituting l-glutamate in hydrochloric acid (HCl) prior to successive dilutions. Nevertheless, HCl, even at low concentrations, may alter the pH of the cell culture medium and interfere with cell activity. Thus, the aim of this study was to evaluate whether the reconstitution of l-glutamate powder in HCl alters its capacity to induce neurotoxicity in different human neuroblastoma cell lines. SH-SY5Y, IMR-32 and SK-N-BE(2) cells were exposed to various concentrations of l-glutamate, which was either reconstituted in HCl (1M) or post re-equilibrated to the pH of the culture medium (7.5). After 24 and 48h of incubation, changes in the cell viability of treated versus untreated cells were evaluated. The effect of an identical amount of HCl present in the l-glutamate dilutions on neuroblastoma cell survival was also investigated. Our data showed that the neurotoxicity of glutamate reconstituted in HCl was comparable to that of HCl alone. Moreover, the pH variations induced by glutamate or HCl in the culture medium were similar. When the pH of the glutamate stock solution was re-equilibrated, l-glutamate induced variation in cell viability to a lower extent and after a longer incubation time. This study demonstrated that HCl used to reconstitute l-glutamate powder might alter the effect of glutamate itself in neuroblastoma cell cultures. Thus, this information might be useful to scientists who use l-glutamate to induce excitotoxicity or to test neuroprotective agents. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Viability and growth of feline preantral follicles in vitro cultured with insulin growth factor and epidermal growth factor supplemented medium.

    Science.gov (United States)

    Alves, A E; Padilha-Nakaghi, L C; Pires-Butler, E A; Apparicio, M; Silva, Nam; Motheo, T F; Vicente, Wrr; Luvoni, G C

    2017-04-01

    In vitro culture of ovarian preantral follicles has emerged as a reproductive technology aimed at obtaining large amount of oocytes for in vitro embryo production. The addition of growth factors (GF) in the in vitro culture of preantral follicles of different species has provided superior results of follicular development, antrum formation and proliferation of granulosa cells. However, there are only few reports regarding the use of these factors on feline preantral follicle in vitro culture. Thus, the aim of this study was to investigate the effect of a combination of IGF-1 and EGF on in vitro viability and growth of preantral follicles and enclosed oocytes collected from domestic cats. A total of 64 follicles characterized by multilayer granulosa cells were isolated and individually cultured for 6 days (T6) in minimum essential medium supplemented with IGF-1+ EGF (100 ng/ml each) or without (control). A higher percentage of follicles were viable after culture with GF than without, and an increase in size when IGF-1+ EGF were added to the medium (170 ± 32.4 μm (T0) vs. 201 ± 22.3 μm (T6); p  .05). These data suggest that the addition of IGF-1 and EGF to the culture medium promotes the in vitro development of preantral follicles of cats. © 2016 Blackwell Verlag GmbH.

  16. The effects of embryo culture media on the birthweight of singletons via fresh or frozen-thawed embryo transfer: a large-scale retrospective study.

    Science.gov (United States)

    Gu, Fang; Deng, Mingfen; Gao, Jun; Wang, Zilian; Ding, Chenhui; Xu, Yanwen; Zhou, Canquan

    2016-09-19

    Embryo culture media used for IVF treatment might affect fetal growth and thus birthweight of the newborns. A retrospective study was conducted in South China using data from 2370 singleton neonates born after IVF/ICSI between 2009 and 2012. Two culture media, i.e., either Vitrolife or SAGE were used as embryo culture media during the study period. Neonates' birthweights were compared between the two embryo culture media groups. Among the 2370 singletons, 1755 cases came from fresh cleavage embryo transfer while 615 were from frozen-thawed cleavage embryo transfer. Within the fresh embryo transfer newborns, no statistical difference was observed in either birthweight (mean ± SD: 3196.0 ± 468.9 versus 3168.4 ± 462.0g, p > 0.05) or adjusted birthweight controlled for gestational age and gender (z-score mean ± SD: 0.11 ± 1.02 versus 0.11 ± 0.99 g, P > 0.05) between the Vitrolife (n = 419) and the SAGE group (n = 1336). Likewise within frozen embryo transfer neotates, no statistical difference of the birthweight (3300.6 ± 441.3 vs.3256.0 ± 466.7 g, P > 0.05) and adjusted birthweight (0.30 ± 0.99 g versus 0.29 ± 0.97 g, P > 0.05) was found between the Vitrolife (n = 202) and the SAGE group (n = 413). The sex ratio [OR1.17, 95 % CI (0.94-1.46)/OR1.1, 95 % CI (0.78-1.54)], rate of small for gestational age [OR1.14, 95 % CI (0.82-1.59)/OR1.06, 95 % CI (0.56-2.02)] and large for gestational age [OR1.07, 95 % CI (0.64-1.76)/OR0.98, 95 % CI (0.47-2.02)] in fresh and frozen-thawed subgourps are all comparable respectively between the two culture media. No group differences were found in the rate of low birthweight and macosomia. Multiple linear regression analysis demonstrated that maternal weight, gestational age, frozen-thawed embryo transfer and infant gender were significantly related to neonatal birthweight (P cultured in SAGE or Vitrolife media after fresh or frozen-thawed cleavage

  17. Using simulation to study difficult clinical issues: prenatal counseling at the threshold of viability across American and Dutch cultures.

    Science.gov (United States)

    Geurtzen, Rosa; Hogeveen, Marije; Rajani, Anand K; Chitkara, Ritu; Antonius, Timothy; van Heijst, Arno; Draaisma, Jos; Halamek, Louis P

    2014-06-01

    Prenatal counseling at the threshold of viability is a challenging yet critically important activity, and care guidelines differ across cultures. Studying how this task is performed in the actual clinical environment is extremely difficult. In this pilot study, we used simulation as a methodology with 2 aims as follows: first, to explore the use of simulation incorporating a standardized pregnant patient as an investigative methodology and, second, to determine similarities and differences in content and style of prenatal counseling between American and Dutch neonatologists. We compared counseling practice between 11 American and 11 Dutch neonatologists, using a simulation-based investigative methodology. All subjects performed prenatal counseling with a simulated pregnant patient carrying a fetus at the limits of viability. The following elements of scenario design were standardized across all scenarios: layout of the physical environment, details of the maternal and fetal histories, questions and responses of the standardized pregnant patient, and the time allowed for consultation. American subjects typically presented several treatment options without bias, whereas Dutch subjects were more likely to explicitly advise a specific course of treatment (emphasis on partial life support). American subjects offered comfort care more frequently than the Dutch subjects and also discussed options for maximal life support more often than their Dutch colleagues. Simulation is a useful research methodology for studying activities difficult to assess in the actual clinical environment such as prenatal counseling at the limits of viability. Dutch subjects were more directive in their approach than their American counterparts, offering fewer options for care and advocating for less invasive interventions. American subjects were more likely to offer a wider range of therapeutic options without providing a recommendation for any specific option.

  18. Effect of co-culture of mesenchymal stem/stromal cells with pancreatic islets on viability and function outcomes: a systematic review and meta-analysis.

    Science.gov (United States)

    de Souza, Bianca Marmontel; Bouças, Ana Paula; Oliveira, Fernanda Dos Santos de; Reis, Karina Pires; Ziegelmann, Patrícia; Bauer, Andrea Carla; Crispim, Daisy

    2017-03-04

    The maintenance of viable and functional pancreatic islets is crucial for successful islet transplantation from brain-dead donors. To overcome islet quality loss during culture, some studies have co-cultured islets with mesenchymal stem/stromal cells (MSC). However, it is still uncertain if MSC-secreted factors are enough to improve islet quality or if a physical contact between MSCs and islets is needed. Therefore, we performed a systematic review and meta-analysis to clarify the effect of different culture contact systems of islets with MSCs on viability and insulin secretion outcomes. Pubmed and Embase were searched. Twenty studies fulfilled the eligibility criteria and were included in the qualitative synthesis and/or meta-analysis. For both outcomes, pooled weighted mean differences (WMD) between islet cultured alone (control group) and the co-culture condition were calculated. Viability mean was higher in islets co-cultured with MSCs compared with islet cultured alone [WMD = 18.08 (95% CI 12.59-23.57)]. The improvement in viability was higher in islets co-cultured in indirect or mixed contact with MSCs than in direct physical contact (P culture condition compared with islet cultured alone [WMD = 0.83 (95% CI 0.54-1.13)], independently of contact system. Results from the studies that were analyzed only qualitatively are in accordance with meta-analysis data. Co-culture of islets with MSCs has the potential for protecting islets from injury during culture period. Moreover, culture time appears to influence the beneficial effect of different methods of co-culture on viability and function of islets.

  19. VIABILITY AND ISOLATION OF MARINE-BACTERIA BY DILUTION CULTURE - THEORY, PROCEDURES, AND INITIAL RESULTS

    NARCIS (Netherlands)

    BUTTON, DK; SCHUT, F; QUANG, P; MARTIN, R; ROBERTSON, BR

    Dilution culture, a method for growing the typical small bacteria from natural aquatic assemblages, has been developed. Each of 11 experimental trials of the technique was successful. Populations are measured, diluted to a small and known number of cells, inoculated into unamended sterilized

  20. Impact of the freeze-drying process on product appearance, residual moisture content, viability, and batch uniformity of freeze-dried bacterial cultures safeguarded at culture collections.

    Science.gov (United States)

    Peiren, Jindrich; Hellemans, Ann; De Vos, Paul

    2016-07-01

    In this study, causes of collapsed bacterial cultures in glass ampoules observed after freeze-drying were investigated as well as the influence of collapse on residual moisture content (RMC) and viability. Also, the effect of heat radiation and post freeze-drying treatments on the RMC was studied. Cake morphologies of 21 bacterial strains obtained after freeze-drying with one standard protocol could be classified visually into four major types: no collapse, porous, partial collapse, and collapse. The more pronounced the collapse, the higher residual moisture content of the freeze-dried product, ranging from 1.53 % for non-collapsed products to 3.62 % for collapsed products. The most important cause of collapse was the mass of the inserted cotton plug in the ampoule. Default cotton plugs with a mass between 21 and 30 mg inside the ampoule did not affect the viability of freeze-dried Aliivibrio fischeri LMG 4414(T) compared to ampoules without cotton plugs. Cotton plugs with a mass higher than 65 mg inside the ampoule induced a full collapsed product with rubbery look (melt-back) and decreasing viability during storage. Heat radiation effects in the freeze-drying chamber and post freeze-drying treatments such as exposure time to air after freeze-drying and manifold drying time prior to heat sealing of ampoules influenced the RMC of freeze-dried products. To produce uniform batches of freeze-dried bacterial strains with intact cake structures and highest viabilities, inserted cotton plugs should not exceed 21 mg per ampoule. Furthermore, heat radiation effects should be calculated in the design of the primary drying phase and manifold drying time before heat sealing should be determined as a function of exposure time to air.

  1. Exposure to Music Alters Cell Viability and Cell Motility of Human Nonauditory Cells in Culture

    Directory of Open Access Journals (Sweden)

    Nathalia R. Lestard

    2016-01-01

    Full Text Available Although music is part of virtually all cultures in the world, little is known about how it affects us. Since the beginning of this century several studies suggested that the response to music, and to sound in general, is complex and might not be exclusively due to emotion, given that cell types other than auditory hair cells can also directly react to audible sound. The present study was designed to better understand the direct effects of acoustic vibrations, in the form of music, in human cells in culture. Our results suggest that the mechanisms of cell growth arrest and/or cell death induced by acoustic vibrations are similar for auditory and nonauditory cells.

  2. Viability of mycelial growth of Coprinus Comatus in culture medium based on organic residues

    OpenAIRE

    de Almeida, Lais Benes; Sales-Campos, Ceci; Melo de Carvalho, Cristiane Suely; de Almeida Minhoni, Marli Teixeira [UNESP; Nogueira de Andrade, Meire Cristina

    2012-01-01

    The objective of this work was to evaluate the mycelial growth of the Coprinus comatus strain CCO 01/01 in culture based on organic residues of Saccharum officinarum (sugarcane bagasse), Citrus sinensis (orange bagasse), Ananas comosus (pineapple residues) and Musa sp. (banana leaf), supplemented with wheat bran in the proportions of 0, 10 and 20%, kept at 27 degrees C. The mycelial growth of C. comatus was evaluated daily by measurement of the diameter of the colony during seven days of incu...

  3. Cat and dog primordial follicles enclosed in ovarian cortex sustain viability after in vitro culture on agarose gel in a protein-free medium.

    Science.gov (United States)

    Fujihara, M; Comizzoli, P; Wildt, D E; Songsasen, N

    2012-12-01

    Our objective was to examine the influences of differing media, protein supplementation and the microenvironment on cat vs dog primordial follicle viability in vitro. Ovarian cortical slices were cultured for 3, 9 or 15 days in α-minimum essential medium (α-MEM) or MEM supplemented with 10% fetal bovine serum (FBS), 10% knock-out serum replacement (KSR) or 0.1% polyvinyl alcohol (protein free). In a separate study, cat and dog ovarian tissues were cultured in protein-free α-MEM and MEM, respectively, in cell culture inserts, on 1.5% agarose gel or in 24-well cell culture plates (control). Follicle viability was assessed in both studies using calcein AM/ethidium homodimer and histological evaluation with haematoxylin/eosin staining. No cat follicle sustained viability beyond 9 days of in vitro culture in α-MEM compared to 37.5% of those incubated for 15 days in MEM in protein-free condition (p free condition at 15 days. Serum was detrimental (p free condition supported cat follicle viability, whereas the latter was superior (p culture insert and control groups after 3 and 9 days of culture. For the cat, the agarose gel better (p culture insert. Therefore, sustaining primordial follicle survival from intracortical ovarian slices requires a different in vitro microenvironment for the cat vs the dog. A key factor to enhancing survival of these early stage follicles in culture appears to be the use of agarose gel, which enhances follicle viability, perhaps by promoting gas exchange. © 2012 Blackwell Verlag GmbH.

  4. Real architecture For 3D Tissue (RAFT™) culture system improves viability and maintains insulin and glucagon production of mouse pancreatic islet cells.

    Science.gov (United States)

    Szebeni, Gabor J; Tancos, Zsuzsanna; Feher, Liliana Z; Alfoldi, Robert; Kobolak, Julianna; Dinnyes, Andras; Puskas, Laszlo G

    2017-04-01

    There is an unmet medical need for the improvement of pancreatic islet maintenance in culture. Due to restricted donor availability it is essential to ameliorate islet viability and graft engraftment. The aim of this study was to compare the standard tissue culture techniques with the advanced Real Architecture For 3D Tissue (RAFT™) culture system in terms of viability and hormone production. Here, we first report that islets embedded in RAFT™ collagen type I advanced tissue culture system maintain their tissue integrity better than in monolayer and suspension cultures. The Calcein violet assay and Annexin V/propidium-iodide staining show higher cell viability in the RAFT™ culture system. Quantitative real-time PCR data showed that RAFT™ increases insulin expression after 18 days in culture compared to traditional methods. Enhanced insulin and glucagon production was further verified by immunofluorescent staining in a time-course manner. These results indicate that RAFT™ tissue culture platform can be a promising tool to maintain pancreatic islet spheroid integrity and culture islets for downstream high throughput pharmacological studies ex vivo.

  5. Fermented sausage production using E. faecium as starter culture: Physicochemical and microbiological profile, sensorial acceptance and cellular viability

    Directory of Open Access Journals (Sweden)

    Catharina Calochi Pires de Carvalho

    2017-09-01

    Full Text Available Fermented sausages are defined as a mixture of lean meat and fat, curing salts, sucrose and spices, stuffed in a natural or artificial casing and submitted to fermentation and air-drying process. Starter culture and ripening process may affect the quality and acceptability of the final product. Current research evaluates the use of Enterococcus faecium as starter culture in fermented sausage production and its physicochemical and microbiological profile during maturation process, coupled to sausage sensory acceptance after ripening. Enterococcus faecium showed 10.9 log CFU g-1 and remained viable after the ripening period with 8.32 log CFU g-1. Fermented sausage was monitored during the ripening period by physicochemical (pH control, water activity and weight loss and microbiological (analysis of coagulase-positive Staphylococcus, coliforms and Salmonella spp. analyses. All tests complied with standards established by Brazilian legislation and did not interfere in final product quality. Results showed that E. faecium was resistant to curing salt and sodium chloride, maintaining its viability during ripening and conferring beneficial effects on fermented sausage technological characteristics. E. faecium also proved to be in vitro resistant to simulate passage through the human digestive tract. Fermented sausage containing E. faecium had better sensory acceptance than commercial sausage evaluated.

  6. Viability and proliferation of L929, tumour and hybridoma cells in the culture media containing sericin protein as a supplement or serum substitute.

    Science.gov (United States)

    Cao, Ting-Ting; Zhang, Yu-Qing

    2015-09-01

    Cell cultures often require the addition of animal serum and other supplements. In this study, silk sericin, a bioactive protein, recovered from the waste of silk floss production was hydrolysed into three pepsin-degraded sericin peptides with different ranges of molecular mass. Normal animal cells, tumour cells and hybridoma cells were cultured systematically in FBS culture media containing sericin as a supplement or serum substitute. The culture test and microscopic observation of L929 cells showed that the smaller molecular weight of the degraded sericin is most suitable for cell culture. The cell culture results showed that with the degradation of sericin, for normal mouse fibroblast L929 cells, addition of 0.75 % sericin into FBS culture medium yields cell viability that is superior to FBS culture medium alone. When all serum was replaced by sericin, cell viability in the sericin medium could reach about one half of that in FBS medium. When in a medium containing a mixture of FBS: sericin (6:4, v/v), the cell culture effect is about 80 %. For the cultures of four tumour and one hybridoma cells, regardless of the molecular weight range, these degraded sericin peptides could substitute all serum in FBS media. The cell viability and proliferation of these tumour and hybridoma cells are equivalent or superior to that in FBS medium. In other words, cell viability and proliferation of these tumour and hybridoma cells in sericin media are more preferable to serum media. The mechanism of the sericin protein to promote cell growth and proliferation will be further investigated later.

  7. Elaboration of fermented dairy beverages: acceptability and viability of probiotic cultures

    Directory of Open Access Journals (Sweden)

    Adriana Carla Santos de Menezes Ramos

    2013-12-01

    Full Text Available This work aims to develop formulations of fermented dairy beverages with probiotic cultures (Lactobacillus acidophilus, Streptococcus thermophilus e Bifidobacterium bifidum yellow monbin flavored, to characterize the chemical composition and to evaluate the stability during refrigerated storage for 28 days.The dairy beverages were developed from an experimental design 3x3, with two factors and three levels each: yellow monbin pulp (15, 20 and 25% and whey (20, 30 and 40%. Nine formulations were produced and submitted to a sensory acceptability test. Thereafter three selected formulations were evaluated based on their microbiological characteristics, proximate composition, pH and titratable acidity. As the acceptance was higher than 70% in all the formulations, were selected that had a higher proportion of whey. Thus, the formulations F3, F6 e F9 were selected. Considering the proximate composition, the selected formulations not differed regarding for carbohydrates. The dairy beverages showed stability with respect to pH and acidity during the shelf life of 28 days and satisfactory results as the investigation of pathogenic microorganisms within the standards established. Quantification of lactic acid bacteria evidenced high numbers to Lactobacillus acidophilus and Streptococcus thermophilus, values between 11.6 to 10.2 CFU log/mL and 8.9 to 11 CFU log/mL, respectively. The species Bifidobacterium bifidum presented less than 6 log/ mL CFU for the formulations. Based on these results, the selected dairy beverages formulations had nutrition, technology and sensorial feasibility.

  8. Comparative analysis of bacterial community composition in bulk tank raw milk by culture-dependent and culture-independent methods using the viability dye propidium monoazide.

    Science.gov (United States)

    Weber, Mareike; Geißert, Janina; Kruse, Myriam; Lipski, André

    2014-11-01

    Microbial diversity of 3 raw milk samples after 72 h of storage at 4 °C in a bulk tank was analyzed by culture-dependent and -independent methods. The culture-dependent approach was based on the isolation of bacteria on complex and selective media, chemotaxonomic differentiation of isolates, and subsequent identification by 16S rRNA gene sequencing. The culture-independent approach included the treatment of raw milk with the dye propidium monoazide before direct DNA extraction by mechanic and enzymatic cell lysis approaches, and cloning and sequencing of the 16S rRNA genes. The selective detection of viable bacteria improved the comparability between bacterial compositions of raw milk based on culture-dependent and -independent methods, which was the major objective of this study. Several bacterial species of the phyla Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria were detected by the culture-dependent method, whereas mainly bacteria of the phylum Proteobacteria as well as low proportions of the phyla Bacteroidetes and Actinobacteria were detected by the culture-independent method. This led to the conclusion that the phylum Firmicutes was strongly discriminated by the culture-independent approach. Generally, species richness detected by the culture-dependent method was higher than that detected by the culture-independent method for all samples. However, few taxa could be detected solely by the direct DNA-based method. In conclusion, the combination of culture-dependent and -independent methods led to the detection of the highest bacterial diversity for the raw milk samples analyzed. It was shown that DNA extraction from raw milk as the essential step in culture-independent methods causes the discrimination of taxa by incomplete cell lysis. Treatment of raw milk with the viability dye propidium monoazide was optimized for the application in raw milk without former removal of milk ingredients and proved to be a suitable tool to ensure comparability

  9. Cry1Ab treatment has no effects on viability of cultured porcine intestinal cells, but triggers Hsp70 expression.

    Directory of Open Access Journals (Sweden)

    Angelika Bondzio

    Full Text Available In vitro testing can contribute to reduce the risk that the use of genetically modified (GM crops and their proteins show unintended toxic effects. Here we introduce a porcine intestinal cell culture (IPEC-J2 as appropriate in vitro model and tested the possible toxic potential of Cry1Ab protein, commonly expressed in GM-maize. For comprehensive risk assessment we used WST-1 conversion and ATP content as metabolic markers for proliferation, lactate dehydrogenase release as indicator for cells with compromised membrane and transepithelial electrical resistance as parameter indicating membrane barrier function. The results were compared to the effects of valinomycin, a potassium ionophore, known to induce cytotoxic effects in most mammalian cell types. Whereas no toxicity was observed after Cry1Ab treatment, valinomycin induced a decrease in IPEC-J2 viability. This was confirmed by dynamic monitoring of cellular responses. Additionally, two dimensional differential in-gel electrophoresis was performed. Only three proteins were differentially expressed. The functions of these proteins were associated with responses to stress. The up-regulation of heat shock protein Hsp70 was verified by Western blotting as well as by enzyme-linked immunosorbent assay and may be related to a protective function. These findings suggest that the combination of in vitro testing and proteomic analysis may serve as a promising tool for mechanism based safety assessment.

  10. Short communication: Viability of culture organisms in honey-enriched acidophilus-bifidus-thermophilus (ABT)-type fermented camel milk.

    Science.gov (United States)

    Varga, L; Süle, J; Nagy, P

    2014-11-01

    The aim of this research was to monitor the survival during refrigerated storage of Lactobacillus acidophilus LA-5 (A), Bifidobacterium animalis ssp. lactis BB-12 (B), and Streptococcus thermophilus CHCC 742/2130 (T) in cultured dairy foods made from camel and, for comparison, cow milks supplemented with black locust (Robinia pseudoacacia L.) honey and fermented by an acidophilus-bifidus-thermophilus (ABT)-type culture. Two liters of dromedary camel milk and 2 L of cow milk were heated to 90 °C and held for 10 min, then cooled to 40 °C. One half of both types of milk was fortified with black locust honey at the rate of 5.0% (wt/vol), whereas the other half was devoid of honey and served as a control. The camel and cow milks with and without honey were subsequently inoculated with ABT-5 culture and were fermented at 37 °C until a pH value of 4.6 was reached. Thereafter, the probiotic fermented milks were cooled to 15 °C in ice water and were each separated into 18 fractions that were transferred in sterile, tightly capped centrifuge tubes. After 24 h of cooling at 8 °C (d 0), the samples were stored at refrigeration temperature (4 °C). Three tubes of all 4 products (i.e., fermented camel and cow milks with and without honey) were taken at each sampling time (i.e., following 0, 7, 14, 21, 28, and 35 d of storage), and the counts of characteristic microorganisms and those of certain spoilage microbes (yeasts, molds, coliforms, Escherichia coli) were enumerated. The entire experimental program was repeated twice. The results showed that addition of black locust honey at 5% to heat-treated camel and cow milks did not influence the growth and survival of starter streptococci during production and subsequent refrigerated storage of fermented ABT milks. In contrast, honey improved retention of viability of B. animalis ssp. lactis BB-12 in the camel milk-based product during storage at 4 °C up to 5 wk. No spoilage organisms were detected in any of the samples tested

  11. Viability Theory

    CERN Document Server

    Aubin, Jean-Pierre; Saint-Pierre, Patrick

    2011-01-01

    Viability theory designs and develops mathematical and algorithmic methods for investigating the adaptation to viability constraints of evolutions governed by complex systems under uncertainty that are found in many domains involving living beings, from biological evolution to economics, from environmental sciences to financial markets, from control theory and robotics to cognitive sciences. It involves interdisciplinary investigations spanning fields that have traditionally developed in isolation. The purpose of this book is to present an initiation to applications of viability theory, explai

  12. Viability and contribution to proteolysis of an adjunct culture of Lactobacillus plantarum in two model cheese systems: cheddar cheese-type and soft-cheese type.

    Science.gov (United States)

    Milesi, M M; McSweeney, P L H; Hynes, E R

    2008-09-01

    The influence of the cheese-making process, ripening conditions and primary starter on the viability and proteolytic activity of an adjunct culture of Lactobacillus plantarum I91 was assessed in two miniature cheese models, representative of Cremoso Argentino and Cheddar cheeses. Cheeses with and without adjunct culture were made under controlled microbiological conditions and sampled during ripening for physicochemical and microbiological analyses. The addition of lactobacilli neither contributed to acid production nor caused changes to the composition of the cheeses. The strain studied exhibited good development and survival and showed a similar growth pattern in both cheese matrices. The adjunct culture caused changes to secondary proteolysis of both cheese types, which were evidenced by modification of peptide profiles and the increase in the levels of some individual amino acids as well as the total content of free amino acids. The changes observed were consistent with the acceleration of proteolysis in the two cheese models assayed. Lactobacillus plantarum I91 has desirable and robust technological properties, which makes it a suitable adjunct culture for cheese-making. Other cultures and environmental conditions prevailing in the food may affect the viability of adjunct cultures and its biochemical activities; this is the first report describing the successful performance of an adjunct culture of Lact. plantarum I91 in two different model cheese systems.

  13. Establishment, maintenance and functional integrity of the blood-testis barrier in organotypic cultures of fresh and frozen/thawed prepubertal mouse testes.

    Science.gov (United States)

    Rondanino, C; Maouche, A; Dumont, L; Oblette, A; Rives, N

    2017-05-01

    Can the spatio-temporal formation of an intact blood-testis barrier (BTB), which is essential for the progression of spermatogenesis, be reproduced in cultures of fresh or frozen/thawed prepubertal mouse testes? Organotypic cultures allow the establishment and maintenance of major BTB components and the formation of a functional BTB in mouse testicular tissues. In vitro maturation of prepubertal testicular tissues is a promising approach to restore fertility in adult survivors of childhood cancer. Although gametes can be successfully obtained from prepubertal mouse testes in organotypic cultures, the spermatogenic yield remains low compared to in vivo controls. Mouse testicular tissues were frozen using controlled slow freezing (CSF) or solid surface vitrification (SSV) procedures. A total of 158 testes (fresh n = 58, CSF n = 58 or SSV n = 42) from 6 to 7 days postpartum (dpp) mice were cultured at 34°C in basal medium (α-MEM, 10% KnockOut Serum Replacement, 5 μg/ml gentamicin) at a gas-liquid interphase (under 20% O2), with or without 10-6 M retinol, for 9, 16 and 30 days. In addition, 32 testes from 6-7, 15-16, 22-23 and 36-37 dpp mice were used as in vivo controls. The mRNA levels of BTB genes (Claudin 3, Claudin 11, Zonula occludens 1 and Connexin-43), germ cell-specific genes (Sal-like protein 4, Kit oncogene, Stimulated by retinoic acid gene 8, Synaptonemal complex protein 3, Transition protein 1 and Protamine 2), markers of Sertoli cell immaturity/maturity (anti-Mullerian hormone, androgen receptor, cyclin-dependent kinase inhibitor 1b) and the androgen-regulated gene Reproductive homeobox 5 (Rhox5) were measured by quantitative RT-PCR (RT-qPCR). The localization of BTB proteins in seminiferous tubules was studied by immunohistochemistry and spermatogenic progression was evaluated histologically. The integrity of the BTB was assessed using a biotin tracer. Modest differences in Claudin 11 (Cldn11), Zonula occludens 1 (Zo-1), Connexin-43 (Cx43

  14. Effect of medium composition on the in vitro culture of bovine pre-antral follicles: morphology and viability do not guarantee functionality.

    Science.gov (United States)

    Rossetto, Rafael; Saraiva, Márcia Viviane Alves; dos Santos, Regiane Rodrigues; da Silva, Cleidson Manoel Gomes; Faustino, Luciana Rocha; Chaves, Roberta Nogueira; Brito, Ivina Rocha; Rodrigues, Giovanna Quintino; Lima, Isadora Machado Teixeira; Donato, Mariana Aragão Matos; Peixoto, Christina Alves; de Figueiredo, José Ricardo

    2013-05-01

    Summary This study investigated the effect of three different culture media (α minimum essential medium (α-MEM), McCoy or TCM199 during the in vitro culture (IVC) of bovine isolated pre-antral follicles. Pre-antral follicles greater than 150 μm in size were isolated and cultured for 0 (control), 8 or 16 days in one of the abovementioned culture media. Follicles were evaluated for survival, growth and antrum formation at days 8 and 16. The results showed that TCM199 was the most suitable medium to preserve follicular viability and ultrastructure, resulting in the highest rates of antrum formation. In conclusion, TCM199 promotes the in vitro development of isolated pre-antral follicles without hampering follicular functionality by sustaining in vitro growth and antrum formation.

  15. Effects of adult stocking density on egg production and viability in cultures of the calanoid copepod Acartia tonsa (Dana)

    DEFF Research Database (Denmark)

    Jepsen, Per Meyer; Andersen, Nikolaj; Holm, Thue

    2007-01-01

    The effect of stocking density of the calanoid copepod Acartia tonsa was evaluated in a 96 h rearing experiment. Possible density-dependent egg production and egg viability were analysed at stocking densities of 100, 200, 300, 400 and 600 adults L−1. Temperature, oxygen saturation and algal conce...

  16. Viability and molecular authentication of Coccidioides immitis strains from culture collection of the Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.

    Science.gov (United States)

    Bezerra, Claudia C F; de Lima, Renata F; Lazera, Márcia S; Wanke, Bodo; Borba, Cíntia M

    2006-01-01

    Twenty Coccidioides immitis strains were evaluated. Only 5 of the 20 strains kept under mineral oil maintained their viability while all 5 subcultures preserved in water remained viable and none of the 13 subcultures kept in soil were viable. A 519 bp PCR product from the csa gene confirmed the identity of the strains.

  17. Improved viability and activity of neutrophils differentiated from HL-60 cells by co-culture with adipose tissue-derived mesenchymal stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Park, Yoon Shin; Lim, Goh-Woon [Department of Pediatrics, Ewha Womans University, School of Medicine, Ewha Medical Research Center, Seoul (Korea, Republic of); Cho, Kyung-Ah; Woo, So-Youn; Shin, Meeyoung [Department of Microbiology, Ewha Womans University, School of Medicine, Ewha Medical Research Center, Seoul (Korea, Republic of); Yoo, Eun-Sun [Department of Pediatrics, Ewha Womans University, School of Medicine, Ewha Medical Research Center, Seoul (Korea, Republic of); Chan Ra, Jeong [Stem Cell Research Center, RNL BIO, Seoul 153-768 (Korea, Republic of); Ryu, Kyung-Ha, E-mail: ykh@ewha.ac.kr [Department of Pediatrics, Ewha Womans University, School of Medicine, Ewha Medical Research Center, Seoul (Korea, Republic of)

    2012-06-22

    Highlights: Black-Right-Pointing-Pointer Neutropenia is a principal complication of cancer treatment. Black-Right-Pointing-Pointer Co-culture of neutrophils with AD-MSC retained cell survival and proliferation and inhibited neutrophil apoptosis under serum starved conditions. Black-Right-Pointing-Pointer AD-MSC increased functions of neutrophil. Black-Right-Pointing-Pointer AD-MSC promoted the viability of neutrophils by enhancing respiratory burst through the expression of IFN-{alpha}, G-CSF, and TGF-{beta}. Black-Right-Pointing-Pointer AD-MSC can be used to improve immunity for neutropenia treatment. -- Abstract: Neutropenia is a principal complication of cancer treatment. We investigated the supportive effect of adipose tissue-derived mesenchymal stem cells (AD-MSCs) on the viability and function of neutrophils. Neutrophils were derived from HL-60 cells by dimethylformamide stimulation and cultured with or without AD-MSCs under serum-starved conditions to evaluate neutrophil survival, proliferation, and function. Serum starvation resulted in the apoptosis of neutrophils and decreased cell survival. The co-culture of neutrophils and AD-MSCs resulted in cell survival and inhibited neutrophil apoptosis under serum-starved conditions. The survival rate of neutrophils was prolonged up to 72 h, and the expression levels of interferon (IFN)-{alpha}, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor, and transforming growth factor (TGF)-{beta} in AD-MSCs were increased after co-culture with neutrophils. AD-MSCs promoted the viability of neutrophils by inhibiting apoptosis as well as enhancing respiratory burst, which could potentially be mediated by the increased expression of IFN-{alpha}, G-CSF, and TGF-{beta}. Thus, we conclude that the use of AD-MSCs may be a promising cell-based therapy for increasing immunity by accelerating neutrophil function.

  18. Phenotypic and Genetic Evaluation of the Influence of Pseudomonas aeruginosa Culture Fractions on the Human Mesenchymal Stem Cells Viability, Apoptotic Pathways and Cytokine Profile.

    Science.gov (United States)

    Holban, Alina Maria; Bleotu, Coralia; Chifiriuc, Mariana Carmen; Lazar, Veronica

    2017-01-01

    The objective of this study was to investigate the effects of P. aeruginosa PAO1 cellular and soluble culture fractions on human mesenchymal stem cells (MSCs) death signaling pathways and cytokine profile. The bone marrow isolated MSCs, incubated for different periods of time with one of the three P. aeruginosa PAO1 culture fractions, i.e. low density whole cultures, heat inactivated bacterial cultures sediments and sterile supernatants, were submitted to the following assays: i) fluorescence microscopy evaluation of cellular morphology and viability; ii) bax, caspase 9, relA and bcl-2 genes expression analysis by qRT-PCR; and iii) quantification of the level of IL-1β, IL-6, IL-8 and IL-10 cytokines released in the MSCs supernatants determined by ELISA. Results were statistically analyzed using the GraphPad In Stat software. The PAO1 whole cultures exhibited the most relevant influences, impacting on MSCs morphology and viability, interfering with apoptotic pathways and significantly stimulating the production of IL-1β and IL-10, while decreasing the production of IL-6 and IL-8. The culture supernatants increased the production of IL-1β and reduced the secretion of all other tested cytokines, while heat-inactivated bacterial cells significantly stimulated both IL-1β and IL-10 production. These data could suggest that in vivo, the fate of P. aeruginosa infection depends on the proportion between different bacterial culture fractions (i.e. the number of viable bacterial cells, the number of dead cells and the amount of bacterial soluble products accumulated locally) that could be influenced by the initial infective dose, by the host defense mechanisms, and also by the administered antimicrobial treatment that may thus interfere with the evolution and magnitude of the induced lesions. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  19. Preliminary viability studies of fibroblastic cells cultured on microcrystalline and nanocrystalline diamonds produced by chemical vapour deposition method

    Directory of Open Access Journals (Sweden)

    Ana Amélia Rodrigues

    2013-02-01

    Full Text Available Implant materials used in orthopedics surgery have demonstrated some disadvantages, such as metallic corrosion processes, generation of wear particles, inflammation reactions and bone reabsorption in the implant region. The diamond produced through hot-filament chemical vapour deposition method is a new potential biomedical material due to its chemical inertness, extreme hardness and low coefficient of friction. In the present study we analysis two samples: the microcrystalline diamond and the nanocrystalline diamond. The aim of this study was to evaluate the surface properties of the diamond samples by scanning electron microscopy, Raman spectroscopy and atomic force microscopy. Cell viability and morphology were assessed using thiazolyl blue tetrazolium bromide, cytochemical assay and scanning electron microscopy, respectively. The results revealed that the two samples did not interfere in the cell viability, however the proliferation of fibroblasts cells observed was comparatively higher with the nanocrystalline diamond.

  20. Preliminary viability studies of fibroblastic cells cultured on microcrystalline and nanocrystalline diamonds produced by chemical vapour deposition method

    Directory of Open Access Journals (Sweden)

    Ana Amélia Rodrigues

    2012-01-01

    Full Text Available Implant materials used in orthopedics surgery have demonstrated some disadvantages, such as metallic corrosion processes, generation of wear particles, inflammation reactions and bone reabsorption in the implant region. The diamond produced through hot-filament chemical vapour deposition method is a new potential biomedical material due to its chemical inertness, extreme hardness and low coefficient of friction. In the present study we analysis two samples: the microcrystalline diamond and the nanocrystalline diamond. The aim of this study was to evaluate the surface properties of the diamond samples by scanning electron microscopy, Raman spectroscopy and atomic force microscopy. Cell viability and morphology were assessed using thiazolyl blue tetrazolium bromide, cytochemical assay and scanning electron microscopy, respectively. The results revealed that the two samples did not interfere in the cell viability, however the proliferation of fibroblasts cells observed was comparatively higher with the nanocrystalline diamond.

  1. Cartilage storage at 4 °C with regular culture medium replacement benefits chondrocyte viability of osteochondral grafts in vitro.

    Science.gov (United States)

    Qi, Jianhong; Hu, Zunjie; Song, Hongqiang; Chen, Bin; Xie, Di; Zhou, Lu; Zhang, Yanming

    2016-09-01

    Maintenance of articular cartilage allografts in culture media is a common method of tissue storage; however, the technical parameters of graft storage remain controversial. In this study, we examined the optimal temperature and culture medium exchange rate for the storage of osteochondral allografts in vitro. Cylindrical osteochondral grafts (n = 120), harvested from the talar joint surface of ten Boer goats, were randomly classified into four groups and stored under the following conditions: Group A1 was maintained at 4 °C in culture medium that was refreshed every 2 days; Group A2 was maintained at 4 °C in the same culture medium, without refreshing; Group B1, was maintained at 37 °C in culture medium that was refreshed every 2 days; Group B2, was maintained at 37 °C in the same culture medium, without refreshing. Chondrocyte viability in the grafts was determined by ethidium bromide/fluorescein diacetate staining on days 7, 21, and 35. Proteoglycan content was measured by Safranin-O staining. Group A1 exhibited the highest chondrocyte survival rates of 90.88 %, 88.31 % and 78.69 % on days 7, 21, and 35, respectively. Safranin O staining revealed no significant differences between groups on days 21 and 35. These results suggest that storage of osteochondral grafts at 4 °C with regular culture medium replacement should be highly suitable for clinical application.

  2. Quantitative evaluation of viability- and apoptosis-related genes in Ascaris suum eggs under different culture-temperature conditions.

    Science.gov (United States)

    Yu, Yong-Man; Cho, You-Hang; Youn, Young-Nam; Quan, Juan Hua; Choi, In-Wook; Lee, Young-Ha

    2012-09-01

    Ascaris suum eggs are inactivated by composting conditions; however, it is difficult to find functional changes in heat-treated A. suum eggs. Here, unembryonated A. suum eggs were incubated at 20°C, 50°C, and 70°C in vitro, and the gene expression levels related to viability, such as eukaryotic translation initiation factor 4E (IF4E), phosphofructokinase 1 (PFK1), and thioredoxin 1 (TRX1), and to apoptosis, such as apoptosis-inducing factor 1 (AIF1) and cell death protein 6 (CDP6), were evaluated by real-time quantitative RT-PCR. No prominent morphological alterations were noted in the eggs at 20°C until day 10. In contrast, the eggs developed rapidly, and embryonated eggs and hatched larvae began to die, starting on day 2 at 50°C and day 1 at 70°C. At 20°C, IF4E, PFK1, and TRX1 mRNA expression was significantly increased from days 2-4; however, AIF1 and CDP6 mRNA expression was not changed significantly. IF4E, PFK1, and TRX1 mRNA expression was markedly decreased from day 2 at 50° and 70°C, whereas AIF1 and CDP6 mRNA expression was significantly increased. The expressions of HSP70 and HSP90 were detected for 9-10 days at 20°C, for 3-5 days at 50°C, and for 2 days at 70°C. Taken together, incremental heat increases were associated with the rapid development of A. suum eggs, decreased expression of genes related to viability, and earlier expression of apoptosis-related genes, and finally these changes of viability- and apoptosis-related genes of A. suum eggs were associated with survival of the eggs under temperature stress.

  3. Frozen spin targets

    CERN Document Server

    Parsons, A S L

    1976-01-01

    Describes six projects which use the frozen-spin principle: Helium-3 R.M.S. and longitudinally polarized frozen spin targets at Rutherford Laboratory, and the frozen spin targets at KEK, Saclay and the one used by the CERN-Helsinki collaboration. (7 refs).

  4. Viability and molecular authentication of Coccidioides spp. isolates from the Instituto de Medicina Tropical de São Paulo culture collection, Brazil

    Directory of Open Access Journals (Sweden)

    Sarah Desirée Barbosa Cavalcanti

    2013-02-01

    Full Text Available Coccidioidomycosis is an emerging fungal disease in Brazil; adequate maintenance and authentication of Coccidioides isolates are essential for research into genetic diversity of the environmental organisms, as well as for understanding the human disease. Seventeen Coccidioides isolates maintained under mineral oil since 1975 in the Instituto de Medicina Tropical de São Paulo (IMTSP culture collection, Brazil, were evaluated with respect to their viability, morphological characteristics and genetic features in order to authenticate these fungal cultures. Only five isolates were viable after almost 30 years, showing typical morphological characteristics, and sequencing analysis using Coi-F and Coi-R primers revealed 99% identity with Coccidioides genera. These five isolates were then preserved in liquid nitrogen and sterile water, and remained viable after two years of storage under these conditions, maintaining the same features.

  5. Viability and molecular authentication of Coccidioides spp. isolates from the Instituto de Medicina Tropical de São Paulo culture collection, Brazil.

    Science.gov (United States)

    Cavalcanti, Sarah Desirée Barbosa; Vidal, Mônica Scarpelli Martinelli; Sousa, Maria da Glória Teixeira de; Del Negro, Gilda Maria Barbaro

    2013-01-01

    Coccidioidomycosis is an emerging fungal disease in Brazil; adequate maintenance and authentication of Coccidioides isolates are essential for research into genetic diversity of the environmental organisms, as well as for understanding the human disease. Seventeen Coccidioides isolates maintained under mineral oil since 1975 in the Instituto de Medicina Tropical de São Paulo (IMTSP) culture collection, Brazil, were evaluated with respect to their viability, morphological characteristics and genetic features in order to authenticate these fungal cultures. Only five isolates were viable after almost 30 years, showing typical morphological characteristics, and sequencing analysis using Coi-F and Coi-R primers revealed 99% identity with Coccidioides genera. These five isolates were then preserved in liquid nitrogen and sterile water, and remained viable after two years of storage under these conditions, maintaining the same features.

  6. Impact of environmental factors on the culturability and viability of Listeria monocytogenes under conditions encountered in food processing plants.

    Science.gov (United States)

    Overney, Anaïs; Jacques-André-Coquin, Joséphine; Ng, Patricia; Carpentier, Brigitte; Guillier, Laurent; Firmesse, Olivier

    2017-03-06

    The ability of Listeria monocytogenes to adhere to and persist on surfaces for months or even years may be responsible for its transmission from contaminated surfaces to food products. Hence the necessity to find effective means to prevent the establishment of L. monocytogenes in food processing environments. The aim of this study was to assess, through a fractional experimental design, the environmental factors that could affect the survival of L. monocytogenes cells on surfaces to thereby prevent the persistence of this pathogen in conditions mimicking those encountered in food processing plants: culture with smoked salmon juice or meat exudate, use of two materials with different hygiene status, biofilm of L. monocytogenes in pure-culture or dual-culture with a Pseudomonas fluorescens strain, application of a drying step after cleaning and disinfection (C&D) and comparison of two strains of L. monocytogenes. Bacterial survival was assessed by culture, qPCR to quantify total cells, and propidium monoazide coupled with qPCR to quantify viable cells and highlight viable but non-culturable (VBNC) cells. Our results showed that failure to apply C&D causes cell persistence on surfaces. Moreover, the sanitation procedure leads only to a loss of culturability and appearance of VBNC populations. However, an additional daily drying step after C&D optimises the effectiveness of these procedures to reduce culturable populations. Our results reinforce the importance to use molecular tools to monitor viable pathogens in food processing plants to avoid underestimating the amounts of cells using only methods based on cell culture. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Comparison of false negative rates and limits of detection following macrofoam-swab sampling of Bacillus anthracis surrogates via Rapid Viability PCR and plate culture.

    Science.gov (United States)

    Hutchison, Janine R; Piepel, Greg F; Amidan, Brett G; Hess, Becky M; Sydor, Michael A; Deatherage Kaiser, Brooke L

    2018-01-21

    We evaluated the effects of Bacillus anthracis surrogates, low surface concentrations, surface materials, and assay methods on false-negative rate (FNR) and limit of detection (LOD 95 ) for recovering Bacillus spores using a macrofoam-swab sampling procedure. Bacillus anthracis Sterne or Bacillus atrophaeus Nakamura spores were deposited over a range of low target concentrations (2 - 500 coupon -1 ) onto glass, stainless steel, vinyl tile, and plastic. Samples were assayed using a modified Rapid Viability-PCR (mRV-PCR) method and the traditional plate culture method to obtain FNR and LOD 95 results. Mean FNRs tended to be lower for mRV-PCR compared to culturing, and increased as spore concentration decreased for all surface materials. Surface material, but not B. anthracis surrogate, influenced FNRs with the mRV-PCR method. The mRV-PCR LOD 95 was lowest for glass and highest for vinyl tile. LOD 95 values overall were lower for mRV-PCR than for the culture method. This study adds to the limited data on FNR and LOD 95 for mRV-PCR and culturing methods with low concentrations of B. anthracis sampled from various surface materials by the CDC macrofoam-swab method. These are key inputs for planning characterization and clearance studies for low contamination levels of B. anthracis. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  8. Short communication: The effects of frozen storage on the survival of probiotic microorganisms found in traditionally and commercially manufactured kefir.

    Science.gov (United States)

    O'Brien, K V; Aryana, K J; Prinyawiwatkul, W; Ordonez, K M Carabante; Boeneke, C A

    2016-09-01

    Kefir is a fermented milk traditionally made from a unique starter culture, which consists of numerous bacteria and yeast species bound together in an exopolysaccharide matrix produced by certain lactic acid bacteria. Many health benefits are associated with traditionally produced kefir; however, bulging and leaking packaging, caused by secondary yeast fermentation during storage, has limited large-scale manufacture. Commercial kefir products have been designed to reduce these effects by using a pure starter culture consisting of a mixture of bacteria and yeast species that give a flavor similar to traditional kefir, but some health benefits may be lost in commercial production due to reduced microbial diversity and lack of beneficial exopolysaccharides. In this study, traditional and commercial kefir was frozen to study the effects of frozen storage on the viability of probiotic bacteria over time. Traditional kefir was prepared by inoculating 1L of pasteurized whole goat milk with approximately 30g of kefir grains. Commercial kefir was prepared by inoculating 1L of full-fat, pasteurized goat milk with a commercial kefir starter. The milk was allowed to ferment at room temperature (24-28°C) until pH 4.6 was reached. Samples were frozen (-8 to -14°C) immediately following the completion of fermentation and were thawed and plated for lactobacilli, lactococci, and yeasts on d 0, 7, 14, and 30 of frozen storage. Lactobacilli, lactococci, and yeasts were significantly reduced in number during frozen storage; however, the traditionally produced kefir was shown to have significantly higher counts of bacteria and yeast at each sampling. We concluded that frozen storage and the development of frozen kefir products could eliminate most packaging concerns associated with the large-scale manufacture of traditionally produced kefir, resulting in increased production and marketability of this healthful product. Copyright © 2016 American Dairy Science Association. Published

  9. Cell viability, proliferation and extracellular matrix production of human annulus fibrosus cells cultured within PDLLA/Bioglass composite foam scaffolds in vitro.

    Science.gov (United States)

    Helen, Wilda; Gough, Julie E

    2008-03-01

    The objective of this study was to assess cell viability, attachment, morphology, proliferation, and collagen and sulphated glycosaminoglycan (s-GAG) production by human annulus fibrosus (HAF) cells cultured in vitro in poly(d,l-lactide) (PDLLA)/Bioglass composite foams. PDLLA foams with different percentages (0, 5 and 30wt.%) of Bioglass particles were prepared by thermally induced phase separation (TIPS) and characterized by scanning electron microscopy (SEM). HAF cell viability in the PDLLA/Bioglass foam was analysed using Live/Dead staining. HAF cell attachment was observed using SEM. An assessment of cell proliferation was conducted using the WST-1 assay. The level of s-GAG and collagen produced by HAF cells was quantified using the 1,9-dimethylmethylene blue (DMMB) assay and Sircoltrade mark assay after 4 weeks of culture. The presence of collagen types I and II within the PDLLA/Bioglass composite foams was analysed using immunohistochemistry. Live/dead staining showed that many viable HAF cells were present on the top surface of the foams as well as penetrating into the internal pore structure, suggesting that the PDLLA/Bioglass composite materials are non-toxic and that the presence of Bioglass particles within PDLLA scaffolds does not inhibit HAF cell growth. The SEM observations revealed that more clusters of HAF cells were attached to the pore walls of both the PDLLA/5BG foam and the PDLLA/30BG foam when compared with the PDLLA/0BG foam. WST-1 assay performed over a period of 4 weeks showed an increased tendency of HAF cells to proliferate within both the PDLLA/5BG foam and the PDLLA/30BG foam when compared with both the tissue culture plastic control and the PDLLA/0BG foam, indicating the presence of Bioglass in the foam has a positive effect on HAF cell proliferation. Sircoltrade mark and DMMB assays showed that HAF cells cultured within the PDLLA/30BG foam had a greater ability to deposit collagen and proteoglycan when compared with the control and

  10. Reduced engraftment and wound closure of cryopreserved cultured skin substitutes grafted to athymic mice.

    Science.gov (United States)

    Harriger, M D; Supp, A P; Swope, V B; Boyce, S T

    1997-09-01

    Cryopreservation of cultured skin substitutes is a requirement for establishment of banks of alternative materials for treatment of acute and chronic skin wounds. To determine whether cryopreservation of skin substitutes that contain cultured cells reduces their efficacy for wound closure, cell-biopolymer grafts were frozen, recovered into culture, and grafted to wounds on athymic mice. Grafts consisted of cultured human keratinocytes and fibroblasts attached to collagen-glycosaminoglycan substrates that were frozen in cell culture medium with 20% serum and 10% DMSO at a controlled rate and stored overnight in liquid nitrogen. After recovery into culture for 24 h, frozen or unfrozen (control) skin substitutes were grafted to full-thickness wounds on athymic mice. Wound area and surface electrical capacitance were measured at 2, 3, and 4 weeks after grafting at which time animals were sacrificed. Wounds were scored for presence of human cells by direct immunofluorescence staining with a monoclonal antibody to HLA-ABC. The data demonstrate that cell-biopolymer grafts are less efficacious after controlled-rate cryopreservation using 10% DMSO as a cryoprotectant. Frozen grafts at 4 weeks after surgery have significantly smaller wound areas, higher capacitance (wetter surface), and fewer healed wounds that contain human cells. The results suggest that these conditions for cryopreservation of cultured grafts reduce graft viability. Improved conditions for cryopreservation are required to maintain viability and efficacy of cultured skin substitutes after frozen storage. Copyright 1997 Academic Press.

  11. Inhibitory effects of multiwall carbon nanotubes with high iron impurity on viability and neuronal differentiation in cultured PC12 cells.

    Science.gov (United States)

    Meng, Li; Jiang, Aihua; Chen, Rui; Li, Chen-zhong; Wang, Liming; Qu, Ying; Wang, Peng; Zhao, Yuliang; Chen, Chunying

    2013-11-08

    The increasing use of carbon nanotubes (CNTs) in biomedical applications has garnered a great concern on their potential negative effects to human health. CNTs have been reported to potentially disrupt normal neuronal function and they were speculated to accumulate and cause brain damage, although a lot of distinct and exceptional properties and potential wide applications have been associated with this material in neurobiology. Fe impurities strapped inside the CNTs may be partially responsible for neurotoxicity generation. In the present study, we selected rat pheochromocytoma (PC12) cells to investigate and compare the effects of two kinds of multiwall carbon nanotubes (MWCNTs) with different concentrations of Fe impurities which usually come from the massive production of CNTs by chemical vapor deposition. Exposure to Fe-high MWCNTs can reduce cell viability and increase cytoskeletal disruption of undifferentiated PC12 cells, diminish the ability to form mature neurites, and then adversely influence the neuronal dopaminergic phenotype in NGF-treated PC-12 cells. The present results highlight the critical role of iron residue in the adverse response to MWCNTs exposure in neural cells. These findings provide useful information for understanding the toxicity and safe application of carbon nanotubes. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  12. Assessing the Viability of Team Learning with Remedial Students in a Lecture Based Japanese Higher Education Culture

    Science.gov (United States)

    Duncan, Brent

    2013-01-01

    Despite a culture with cooperation as a core value, (Nagao, Takashi, & Okuda, 2011) Japanese higher education generally uses rigid lecture-test teaching models that neither support nor condone small-group learning methods in the classroom. As a result, Japanese college students usually work outside the classroom to develop the collaborative…

  13. Effects of sanitation, freezing and frozen storage on enteric viruses in berries and herbs.

    Science.gov (United States)

    Butot, S; Putallaz, T; Sánchez, G

    2008-08-15

    Norovirus (NV) and hepatitis A virus (HAV) are foodborne enteric viruses associated with outbreaks of disease following consumption of fresh or frozen produce. Model experiments were performed to determine the effectiveness of certain commercial processes for the removal of enteric viruses that might be present in berries and herbs. The survival and persistence of HAV, NV, rotavirus (RV) and feline calicivirus (FCV), a surrogate for NV, in frozen produce over time were determined. Survival and inactivation of HAV, RV and FCV were assessed by viral culture and quantitative reverse transcription-PCR (RT-PCR), whereas NV persistence was determined by quantitative RT-PCR only. Freezing did not significantly reduce the viability of any of the viruses except the infectivity of FCV in strawberries. Frozen storage for 3 months had limited effects on HAV and RV survival in all tested food products, whereas in frozen raspberries and strawberries FCV infectivity showed the highest decay rate due to acid pH. To simulate postharvesting conditions, fresh berries and herbs were rinsed with tap, warm or chlorinated water or with a chlorine dioxide (ClO(2)) solution. Available chlorine at a concentration of 200 ppm and ClO(2) at 10 ppm reduced measurable enteric viruses in raspberry and parsley samples by less than 2 log(10) units.

  14. Apoptosis-resistant E1B-19K-expressing NS/0 myeloma cells exhibit increased viability and chimeric antibody productivity under perfusion culture conditions.

    Science.gov (United States)

    Mercille, S; Massie, B

    1999-06-05

    We have shown previously that recombinant NS/0 myelomas expressing sufficient amounts of E1B-19K were resistant to apoptosis occurring in the late phase of batch culture and under stressful conditions such as cultivation in glutamine-free medium or following heat shock. However, no significant increase in monoclonal antibodies (MAb) was observed during the prolonged stationary phase of these batch cultures. Here, we show that E1B-19K can enhance cell survival and improve MAb productivity in high cell density perfusion culture. Typically, lymphoid cells grown under steady state in perfusion exhibit decreasing viabilities with concomitant accumulation of apoptotic cells. By modulating the ability of these cells to resist to induction of apoptosis in low nutrient environment, a 3-fold decrease in specific death rate from 0.22 day-1 for NS/0 control to 0.07 day-1 for E1B-19K cells was achieved, resulting in a significant improvement in cell viability throughout perfusion. E1B-19K cells at the perfusion plateau phase also exhibited a 3-fold reduction in specific growth rate concomitant with a lower percentage of S and higher percentage of G1 phase cells. This was associated with a 40% decrease in specific oxygen consumption rate, likely related to a reduction in the specific consumption rates of limiting nutrient(s). Expression of E1B-19K consequently had a significant impact on the steady-state viable cell density, allowing maintenance of 11.5 x 10(6) E1B-19K cells/mL versus 5.9 x 10(6) control NS/0 cells/mL for the same amount of fresh medium brought into the system (half a volume per day). Whereas MAb concentrations found in perfusion culture of control NS/0 myelomas were almost 3-fold higher than those found in batch culture; in the case of E1B-19K-expressing myelomas, the MAb concentration in perfusion was more than 7-fold higher than in batch. This was attributable to the 2-fold increase in viable cell plateau and to a 40% increase in the perfusion to batch ratio

  15. Cultures of human colonic epithelial cells isolated from endoscopical biopsies from patients with inflammatory bowel disease. Effect of IFNgamma, TNFalpha and IL-1beta on viability, butyrate oxidation and IL-8 secretion

    DEFF Research Database (Denmark)

    Pedersen, G; Saermark, T; Bendtzen, K

    2000-01-01

    Cytokine-mediated impairment of viability and metabolic function of epithelial cells has been suggested as a possible early pathogenic event in the development of inflammatory bowel disease (IBD). It is currently unknown whether pro-inflammatory cytokines have a direct effect on human...... nontransformed colonic epithelial cells. We investigated the effects of TNFalpha, IFNgamma and IL-1beta on viability, short chain fatty acid (butyrate) oxidation and IL-8 secretion in human colonic epithelial cell cultures in vitro obtained from macroscopically normal mucosa from IBD patients and controls...... decreased to median 68% of unexposed cultures (P Cells from IBD patients were significantly less sensitive...

  16. Quantitative Analyses of Synergistic Responses between Cannabidiol and DNA-Damaging Agents on the Proliferation and Viability of Glioblastoma and Neural Progenitor Cells in Culture.

    Science.gov (United States)

    Deng, Liting; Ng, Lindsay; Ozawa, Tatsuya; Stella, Nephi

    2017-01-01

    Evidence suggests that the nonpsychotropic cannabis-derived compound, cannabidiol (CBD), has antineoplastic activity in multiple types of cancers, including glioblastoma multiforme (GBM). DNA-damaging agents remain the main standard of care treatment available for patients diagnosed with GBM. Here we studied the antiproliferative and cell-killing activity of CBD alone and in combination with DNA-damaging agents (temozolomide, carmustine, or cisplatin) in several human GBM cell lines and in mouse primary GBM cells in cultures. This activity was also studied in mouse neural progenitor cells (NPCs) in culture to assess for potential central nervous system toxicity. We found that CBD induced a dose-dependent reduction of both proliferation and viability of all cells with similar potencies, suggesting no preferential activity for cancer cells. Hill plot analysis indicates an allosteric mechanism of action triggered by CBD in all cells. Cotreatment regimens combining CBD and DNA-damaging agents produced synergistic antiproliferating and cell-killing responses over a limited range of concentrations in all human GBM cell lines and mouse GBM cells as well as in mouse NPCs. Remarkably, antagonistic responses occurred at low concentrations in select human GBM cell lines and in mouse GBM cells. Our study suggests limited synergistic activity when combining CBD and DNA-damaging agents in treating GBM cells, along with little to no therapeutic window when considering NPCs. Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.

  17. Combined effect of a nanoemulsion of D-limonene and nisin on Listeria monocytogenes growth and viability in culture media and foods.

    Science.gov (United States)

    Maté, Javier; Periago, Paula M; Palop, Alfredo

    2016-03-01

    The present work evaluated the antibacterial effect of nanoemulsions from natural compounds d-limonene and nisin against Listeria monocytogenes in tryptic soy broth growth medium, chicken broth, and vegetable cream. Experiments were performed by means of optical density growth curves and studies of viability in culture media and foods. Stability of nanoemulsions was evaluated by a Mastersizer 2000 equipment. Results showed greater effectiveness when applying D-limonene in form of nanoemulsion than when applying it directly, and when using both compounds together, both directly and in the form of nanoemulsion. Concentration of L. monocytogenes decreased at least in three log cycles in all the culture media and foods used within the first 90 min after the addition of the natural antimicrobials combined in form of nanoemulsion. Moreover, the growth of L. monocytogenes was inhibited with the combination of antimicrobials in the four weeks that the experiment lasted. Nanoemulsion technology would solve present problems of solubility and stability of oily antimicrobials in the food industry. © The Author(s) 2015.

  18. The oil-resin of the tropical rainforest tree Copaifera langsdorffii reduces cell viability, changes cell morphology and induces cell death in human endometriotic stromal cultures.

    Science.gov (United States)

    Henriques da Silva, Julianna; Borges, Vinicius Raphael de Almeida; Pereira, Leonardo da Cunha Boldrini; Ferrari, Renato; de Mattos, Rômulo Medina; Barros, Eliane Gouveia de Oliveira; Palmero, Celia Yelimar; Fernandes, Patricia Dias; de Carvalho, Patricia Ribeiro; Pereira de Sousa, Valeria; Cabral, Lucio Mendes; Nasciutti, Luiz Eurico

    2015-12-01

    The hormonal treatment for endometriosis frequently fails to completely eradicate endometriotic implants. A new therapeutic treatment is needed. This study investigates the in-vitro effect of Copaifera langsdorffii oil-resin on human eutopic and ectopic endometrium stromal cell cultures (EuESCs and EctESCs). A nanocomposite system containing the copaiba oil-resin (NanoCOR) was developed and acute toxicity test was performed. Endometrial stromal cells (ESCs) from non-endometriotics controls (CESCs), EuESCs and EctESCs were isolated and treated with different concentrations of NanoCOR, at different time intervals to evaluate its effect on cell morphology, proliferation, viability, necrosis and apoptosis induction. When treated with 50 μg/ml of NanoCOR, the morphology of EctESCs changed, as the actin microfilaments were disorganized, disassembled or disrupted. Moreover, at 24 h of treatment with NanoCOR, the EctESCs viability was inhibited, and a significant number of these cells underwent apoptosis. In EuESCs, these effects were observed only at 48 h. Finally, the treatment of EctESCs with NanoCOR increased the lactate dehydrogenase release into the extracellular medium more than in EuESCs. Our data indicate that NanoCOR has a greater impact on the behaviour of human endometriotic stromal cells than on the eutopic endometrium stromal cells, supporting the idea that NanoCOR should be further investigated as a novel and valuable alternative to treat endometriosis. © 2015 Royal Pharmaceutical Society.

  19. Culture viability of Sardina pilchardus (Fish, Teleost: Preliminary results of growth in captivity up to 18 months

    Directory of Open Access Journals (Sweden)

    José Iglesias

    2014-09-01

    Full Text Available Larvae of Sardina pilchardus were obtained in captivity from fertilized eggs captured in the wild and grown in a 10000-L tank. Isochrysis galbana, Artemia franciscana nauplii and live zooplankton were used as prey during the first two weeks; afterwards, the animals were fed on Artemia metanauplii enriched with Isochrysis galbana. A dry feed (Gemma 0.4 and 0.8 from Skretting S.A. (Burgos, España was supplied from the 3rd to the 18th month. The total length reached by sardines at one year of life was 162.02±9.49 mm, corresponding to a wet weight of 36.12±10.82 g. Total length of the last survivor individual at 18 months was 182.37 mm. An 18-month experiment of sardine culture is described for the first time, and the growth data reported can help to determine its potential as a candidate for marine aquaculture.

  20. Production of a Functional Frozen Yogurt Fortified with Bifidobacterium spp.

    Science.gov (United States)

    Muhammad, Zafarullah; Zhang, Qiu-Xue; Zhu, Zong-Tao

    2017-01-01

    Frozen dairy products have characteristics of both yogurt and ice cream and could be the persuasive carriers of probiotics. Functions of the frozen yogurt containing viable bifidobacterial cells are recognized and favored by the people of all ages. We developed a kind of yogurt supplemented by Bifidobacterium species. Firstly, five strains of Bifidobacterium spp. (Bifidobacterium bifidum ATCC 11547, Bifidobacterium longum ATCC 11549, Bifidobacterium infantis ATCC 11551, Bifidobacterium adolescentis ATCC 11550, and Bifidobacterium breve ATCC 11548) were evaluated based on the feasibility criteria of probiotics, comprising acid production, bile tolerance, and adhesion to epithelial cells. Formerly, we combined the optimum strains with yogurt culture (Lactobacillus delbrueckii subsp. bulgaricus EMCC 11102 and Streptococcus salivarius subsp. thermophilus EMCC 11044) for producing frozen yogurt. Finally, physiochemical properties and sensory evaluation of the frozen yogurt were investigated during storage of 60 days at −18°C. Results directed that Bifidobacterium adolescentis ATCC 11550 and Bifidobacterium infantis ATCC 11551 could be utilized with yogurt culture for producing frozen yogurt. Moreover, the frozen yogurt fermented by two bifidobacterial strains and yogurt culture gained the high evaluation in the physiochemical properties and sensory evaluation. In summary, our results revealed that there was no significant difference between frozen yogurt fermented by Bifidobacterium spp. and yogurt culture and that fermented by yogurt culture only. PMID:28691028

  1. Production of a Functional Frozen Yogurt Fortified with Bifidobacterium spp.

    Directory of Open Access Journals (Sweden)

    Amro Abdelazez

    2017-01-01

    Full Text Available Frozen dairy products have characteristics of both yogurt and ice cream and could be the persuasive carriers of probiotics. Functions of the frozen yogurt containing viable bifidobacterial cells are recognized and favored by the people of all ages. We developed a kind of yogurt supplemented by Bifidobacterium species. Firstly, five strains of Bifidobacterium spp. (Bifidobacterium bifidum ATCC 11547, Bifidobacterium longum ATCC 11549, Bifidobacterium infantis ATCC 11551, Bifidobacterium adolescentis ATCC 11550, and Bifidobacterium breve ATCC 11548 were evaluated based on the feasibility criteria of probiotics, comprising acid production, bile tolerance, and adhesion to epithelial cells. Formerly, we combined the optimum strains with yogurt culture (Lactobacillus delbrueckii subsp. bulgaricus EMCC 11102 and Streptococcus salivarius subsp. thermophilus EMCC 11044 for producing frozen yogurt. Finally, physiochemical properties and sensory evaluation of the frozen yogurt were investigated during storage of 60 days at −18°C. Results directed that Bifidobacterium adolescentis ATCC 11550 and Bifidobacterium infantis ATCC 11551 could be utilized with yogurt culture for producing frozen yogurt. Moreover, the frozen yogurt fermented by two bifidobacterial strains and yogurt culture gained the high evaluation in the physiochemical properties and sensory evaluation. In summary, our results revealed that there was no significant difference between frozen yogurt fermented by Bifidobacterium spp. and yogurt culture and that fermented by yogurt culture only.

  2. Production of a Functional Frozen Yogurt Fortified withBifidobacteriumspp.

    Science.gov (United States)

    Abdelazez, Amro; Muhammad, Zafarullah; Zhang, Qiu-Xue; Zhu, Zong-Tao; Abdelmotaal, Heba; Sami, Rokayya; Meng, Xiang-Chen

    2017-01-01

    Frozen dairy products have characteristics of both yogurt and ice cream and could be the persuasive carriers of probiotics. Functions of the frozen yogurt containing viable bifidobacterial cells are recognized and favored by the people of all ages. We developed a kind of yogurt supplemented by Bifidobacterium species. Firstly, five strains of Bifidobacterium spp. ( Bifidobacterium bifidum ATCC 11547, Bifidobacterium longum ATCC 11549, Bifidobacterium infantis ATCC 11551, Bifidobacterium adolescentis ATCC 11550, and Bifidobacterium breve ATCC 11548) were evaluated based on the feasibility criteria of probiotics, comprising acid production, bile tolerance, and adhesion to epithelial cells. Formerly, we combined the optimum strains with yogurt culture ( Lactobacillus delbrueckii subsp. bulgaricus EMCC 11102 and Streptococcus salivarius subsp. thermophilus EMCC 11044) for producing frozen yogurt. Finally, physiochemical properties and sensory evaluation of the frozen yogurt were investigated during storage of 60 days at -18°C. Results directed that Bifidobacterium adolescentis ATCC 11550 and Bifidobacterium infantis ATCC 11551 could be utilized with yogurt culture for producing frozen yogurt. Moreover, the frozen yogurt fermented by two bifidobacterial strains and yogurt culture gained the high evaluation in the physiochemical properties and sensory evaluation. In summary, our results revealed that there was no significant difference between frozen yogurt fermented by Bifidobacterium spp. and yogurt culture and that fermented by yogurt culture only.

  3. Aceitação sensorial e caracterização de frozen yogurt de leite de cabra com adição de cultura probiótica e prebiótico Sensorial acceptance and characterization of goat's milk frozen yogurt with addition of probiotic culture and prebiotic

    Directory of Open Access Journals (Sweden)

    Larissa de Lima Alves

    2009-12-01

    Full Text Available O leite de cabra é recomendado para pessoas intolerantes ao leite de vaca e/ou portadoras de desordens gastrintestinais. frozen yogurt elaborado a partir de iogurte de leite de cabra foi acrescido de 1% de prebiótico (inulina, 0,25% de cultura starter (Streptococcus thermophilus e Lactobacillus bulgaricus e 0,25% de cultura probiótica (Bifidobacterium animalis e Lactobacillus acidophilus para verificar seu potencial probiótico (após sete e 120 dias de armazenamento, sua qualidade físico-química e sua aceitação sensorial. Durante a estocagem, houve redução de células viáveis da flora microbiana, principalmente dos probióticos. Ainda assim, o produto apresenta potencialidade probiótica pela soma de B. animalis e L. acidophilus atingir o mínimo desejável até o fim do armazenamento. Os valores de extrato seco total (26,6±0,2%, proteína (3,0±0,2, gordura (2,6±0,1%, pH (5,5±0,01 e ácido láctico (0,2±0,01% encontram-se semelhantes aos relatados em outros trabalhos para o mesmo produto. A avaliação sensorial foi realizada por 50 provadores não treinados em teste com escala hedônica de sete níveis e intenção de compra. O produto recebeu bons escores para aparência global, cor, aroma e sabor; já textura e sabor residual obtiveram menores índices de aceitação. Na intenção de compra, 61% dos avaliadores declararam que comprariam o produto, contra 39% que não comprariam. Nessas condições, demonstra-se a viabilidade de elaboração de frozen yogurt de leite caprino acrescido de pre- e probióticos.Goat's milk is recommended for people afflicted with cow milk intolerance and gastro-intestinal disorders. frozen yogurt was elaborated with 1% prebiotic (inulin, 0.25% starter culture (Streptococcus thermophilus and Lactobacillus bulgaricus and 0.25% probiotic culture (Bifidobacterium animalis and Lactobacillus acidophilus to verify probiotic potential (after 7 and 120 days of storage, physical-chemical quality and

  4. Ketahanan dan viabilitas probiotik bakteri asam laktat selama proses pembuatan kultur kering dengan metode freeze dan spray drying [Survival and Viability of Lactid Acid Bacteria Probiotic during production of Dried Culture Using Freeze and Spray Drying Methods

    Directory of Open Access Journals (Sweden)

    Erni Harmayani 1

    2001-08-01

    Full Text Available Selection on 36 lactid aid bacteria isolate from various source (dadih, sausage, infant faces, gato, chinese leaf pickle, growol and yoghurt has been carried out based on their potency to reduce choresterol. Based on their ability to assimilate choresterol, conjugate bile solt, restency on bile salt and low pH, three isolates i.e. Lactobacillus sp. Dad 13, L. asidophillus D2 and L. plantarum Mut 7 have been chosen for further study. Viability of selected cultures during biomass production using coconut water with addition of 0.5% yeast extract, and during production of dried starter culture using freeze and spray dried were investigated. The results show that the growth patern of the three isolates selected were almost similar i.e. reaching maximum amount after 16 hours fermentation at 37°C. biomass production using coconut water produced 109 cfu/ml after 16-18 hours incubation at 37°C. decrease on viability after drying using freeze drier ranged between 0.5-2 log cycles, while that of storage of freeze dried culture during 4 weeks at -20°C caused descreasing in viability of 26-56%.

  5. Viability of Staphylococcus xylosus isolated from artisanal sausages for application as starter cultures in meat products Viabilidade de Staphylococcus xylosus isoladas de embutidos artesanais para aplicação como cultivos iniciadores em produtos cárneos

    Directory of Open Access Journals (Sweden)

    Ângela Maria Fiorentini

    2009-03-01

    Full Text Available Viability of Staphylococcus xylosus isolated from artisanal sausages for application as starter cultures in meat products Viability of Staphylococcus xylosus strains AD1 and U5isolated from natural fermented sausages was investigated as starter cultures in fermented sausages produced in the South Region of Brazil. The study demonstrated that the Staphylococcus xylosus strains AD1 and U5showed significant growth during fermentation, stability over freeze-dried process, negative reaction for staphylococcal enterotoxins and viability for using as a single-strain culture or associated with lactic acid bacteria for production of fermented sausages.Investigamos a viabilidade de cepas de Staphylococcus xylosus (AD1 e U5 isoladas de embutidos com fermentação natural, para aplicação como cultivos iniciadores em embutidos fermentados produzidos na Região Sul do Brasil. O estudo demonstrou que cepas de Staphylococcus xylosus (AD1 e U5 apresentaram crescimento significativo durante a fermentação, estabilidade no processo de liofilização e conservação, ausência de produção de enterotoxinas e viabilidade para aplicação como cultivo iniciador simples ou associado com bactérias lácticas na elaboração de embutidos fermentados.

  6. Influence of culture media in viability test of conidia of entomopathogenic fungi Influência de meios de cultura em teste de viabilidade de fungos entomopatogênicos

    Directory of Open Access Journals (Sweden)

    Edimara Aparecida Francisco

    2006-08-01

    Full Text Available This work aimed at investigatimg whether the culture medium used in viability tests affects the conidial germination of Lecanicillium lecanii, Beauveria bassiana and Paecilomyces fumosoroseus isolates. The tests were performed on microscope slides containing one of the culture media: agar-water (AW, minimal medium (MM, potato-dextrose agar (PDA, potato-dextrose-1% yeast extract agar (PDAY, Sabouraud-dextrose-yeast extract agar (SDAY, and complete medium (CM. Three areas per slide were delimited and 0.05ml of a 5.5 x 105 conidia ml-1 suspension was applied to each area. One bioassay was performed for each isolate. Germination was determined after 15 hours of incubation at 26±0.5°C. The culture media influenced the germination of the species studied, verifying within and inter specific variations. CM and PDA provided the highest germination of L. lecanii isolates and the lowest was obtained on SDAY and AW. The germination of B. bassiana isolates was favoured by CM, PDA and PDAY media, a fact not observed in AW and MM. P. fumosoroseus isolates showed the highest germination on CM and PDA media and the lowest on SDAY. However, some isolates presented high germination on nutrient-poor media (AW and MM.O presente trabalho objetivou investigar se meios de cultura utilizados em teste de viabilidade afetam a germinação de conídios de cinco isolados de Lecanicillium lecanii, cinco de Beauveria bassiana e quatro de Paecilomyces fumosoroseus. Os testes foram realizados em lâminas de microscopia contendo um dos seguintes meios de cultura: Ágar-água (AA, Meio Mínimo (MM, Batata, dextrose e ágar (BDA, Batata, dextrose, ágar e 1% de extrato de levedura (BDAL, Sabouraud, dextrose, ágar e extrato de levedura (SDAL e Meio Completo (MC. Delimitaram-se três áreas por lâmina e em cada uma aplicou-se 0,05mL de uma suspensão com concentração de 5,5 x 105 conídios ml-1. Para cada isolado foi realizado um bioensaio, com seis tratamentos e cinco repeti

  7. Estudo da viabilidade econômica do cultivo da babosa (aloe vera L. Study of economic viability of the Aloe vera L. culture

    Directory of Open Access Journals (Sweden)

    Dionizio Bernardino Bach

    2007-08-01

    Full Text Available Conduziu-se este este trabalho, com o objetivo de estudar a viabilidade econômica da babosa e verificar se o cultivo desta planta é ou não, uma alternativa economicamente viável ao produtor rural. Os dados relativos ao custo de produção, utilizados neste trabalho, foram obtidos junto às áreas de produção e instalações industriais da NATURAMA - Indústria e Comércio de Produtos Agropecuários Ltda, estabelecida no município de Paulo Lopes, SC. Na determinação do custo de produção de um hectare de babosa, em cultivo orgânico, foram consideradas todas as etapas, desde a obtenção das mudas até a conclusão do processamento, quando o produto final encontra-se apto ao consumo, e neste caso, é uma bebida em forma de suco. O processamento dos dados foi realizado utilizando uma planilha eletrônica desenvolvida especificamente para esse fim, a qual estimou os custos utilizando as metodologias do custo total (custos fixos e variáveis e do custo operacional total (custo operacional efetivo e depreciações. Pela análise da rentabilidade, conclui-se que a atividade é economicamente viável.The aim of this work is to study the economic viability of Aloe vera L. and verify if the culture of this plant is or not an alternative economically viable for the farmer. The data of production cost used in this work were gotten in the areas of production and industrial installations of NATURAMA- Industry and Trade of Farming Products LTDA, settled in Paulo Lopes, SC. In the determination of the production cost of one Aloe vera L hectare, all the phases were taken into consideration, from the getting of molts up to the conclusion of the processing, when the final product is ready to the consumption, and in this case, it's a beverage in a way of juice. The processing data was made by using an electronic spread sheet developed specifically for this reason, which estimated the costs using the methodologies of the total cost (steady and variable

  8. Effects of fatty acid-free bovine serum albumin and fetal calf serum supplementing repair cultures on pre- and post-warm viability of biopsied bovine embryos produced in vitro.

    Science.gov (United States)

    Yotsushima, Kenji; Sakaguchi, Minoru; Shimizu, Manabu; Okimura, Tomoko; Izaike, Yoshiaki

    2004-08-01

    The objective of this study was to investigate the influence of fatty acid-free bovine serum albumin (BSA) or fetal calf serum (FCS) on the re-expansion of biopsied blastocysts and post-warm viability of subsequently vitrified embryos. Firstly, blastocysts produced in vitro were biopsied at Day 7 and cultured to allow repair in TCM199 with 0.3% BSA or 5% FCS for 24 h. The re-expansion rates and mean total numbers of cells of the re-expanded embryos after the repair culture with BSA were almost the same as that with FCS. Secondly, after biopsied embryos were similarly cultured for repair with BSA or FCS, re-expanded embryos were selected for vitrification. After warming and exposure to 0.5 M sucrose with 20% FCS in mPBS, the embryos were cultured in TCM199 with 5% FCS for 24 h. The re-expansion rate and mean total number of cells in re-expanded blastocysts in the BSA treatment group (97.4 +/- 2.9% and 106 +/- 42) was significantly higher than that in the FCS treatment group (51.6 +/- 9.1% and 61 +/- 38), respectively (Pbovine biopsied blastocysts; but, compared with BSA supplementation, FCS supplementation during repair culture reduces the post-warm viability of biopsied and subsequently vitrified embryos.

  9. The effects of short-term hypoxia on human mesenchymal stem cell proliferation, viability and p16INK4A mRNA expression: Investigation using a simple hypoxic culture system with a deoxidizing agent

    Directory of Open Access Journals (Sweden)

    Ito A

    2015-05-01

    Full Text Available A hypoxic environment is thought to be important for the maintenance of stemness and suppressing cell senescence, in stem cells. Therefore, a hypoxic condition is induced during cell expansion and/or induction of intended differentiation. However, the induction of these conditions requires a specially equipped hypoxia chamber and expensive gas mixtures, which are expensive and space-consuming. Owing to these restrictions, appropriate hypoxic conditions cannot be provided during cell transportation, which is increasingly required for regenerative medicine. Hence, a simple and economical culture system is required. The purpose of this study was to investigate the effects of short-term hypoxic conditions on human mesenchymal stem cell (MSC proliferation, viability, and senescence, utilizing the CulturePal system (CulturePal-Zero and CulturePal-Five, a novel and simple hypoxic culture system with a built-in deoxidizing agent. The O2 concentration in the CulturePal-Zero was observed to reduce to <0.1% within 1 h, and to 5% within 24 h in the CulturePal-Five system. Cell proliferation under these hypoxic conditions showed a sharp increase at 5% O2 concentration, and no noticeable cell death was observed even at severe hypoxic conditions (<0.1% O2 up to 72h. The p16INK4A (cell senescence marker mRNA expression was retained under hypoxic conditions up to 72h, but it was up-regulated under normoxic conditions. Interestingly, the p16INK4A expression altered proportionately to the O2 concentration. These results indicated that the short-term hypoxic condition, at an approximate O2 concentration of 5%, would be suitable for promoting cell proliferation and repressing cell senescence, without aggravating the MSC viability. Therefore, the CulturePal systems may be suitable for providing an appropriate hypoxic condition in stem cell research and transportation.

  10. The effects of short-term hypoxia on human mesenchymal stem cell proliferation, viability and p16INK4A mRNA expression: Investigation using a simple hypoxic culture system with a deoxidizing agent

    Science.gov (United States)

    Ito, Akira; Aoyama, Tomoki; Yoshizawa, Makoto; Nagai, Momoko; Tajino, Junichi; Yamaguchi, Shoki; Iijima, Hirotaka; Zhang, Xiangkai; Kuroki, Hiroshi

    2015-01-01

    A hypoxic environment is thought to be important for the maintenance of stemness and suppressing cell senescence, in stem cells. Therefore, a hypoxic condition is induced during cell expansion and/or induction of intended differentiation. However, the induction of these conditions requires a specially equipped hypoxia chamber and expensive gas mixtures, which are expensive and space-consuming. Owing to these restrictions, appropriate hypoxic conditions cannot be provided during cell transportation, which is increasingly required for regenerative medicine. Hence, a simple and economical culture system is required. The purpose of this study was to investigate the effects of short-term hypoxic conditions on human mesenchymal stem cell (MSC) proliferation, viability, and senescence, utilizing the CulturePal system (CulturePal-Zero and CulturePal-Five), a novel and simple hypoxic culture system with a built-in deoxidizing agent. The O2 concentration in the CulturePal-Zero was observed to reduce to <0.1% within 1 h, and to 5% within 24h in the CulturePal-Five system. Cell proliferation under these hypoxic conditions showed a sharp increase at 5% O2 concentration, and no noticeable cell death was observed even at severe hypoxic conditions (<0.1% O2) up to 72h. The p16INK4A (cell senescence marker) mRNA expression was retained under hypoxic conditions up to 72h, but it was up-regulated under normoxic conditions. Interestingly, the p16INK4A expression altered proportionately to the O2 concentration. These results indicated that the short-term hypoxic condition, at an approximate O2 concentration of 5%, would be suitable for promoting cell proliferation and repressing cell senescence, without aggravating the MSC viability. Therefore, the CulturePal systems may be suitable for providing an appropriate hypoxic condition in stem cell research and transportation. PMID:26195892

  11. Frozen waves: experimental generation

    OpenAIRE

    Vieira, TA; Gesualdi, MRR; Zamboni-Rached, M

    2012-01-01

    Frozen waves (FWs) are very interesting particular cases of nondiffracting beams whose envelopes are static and whose longitudinal intensity patterns can be chosen a priori. We present here for the first time (that we know of) the experimental generation of FWs. The experimental realization of these FWs was obtained using a holographic setup for the optical reconstruction of computer generated holograms (CGH), based on a 4-f Fourier filtering system and a nematic liquid crystal spatial light ...

  12. Effect of refrigeration systems upon frozen bull sperm viability assessed by computer-assisted sperm analysis and fluorescent probesEfeito de sistemas de refrigeração sobre a viabilidade do sêmen bovino congelado analisado por meio de sistema computadorizado e sondas fluorescentes

    Directory of Open Access Journals (Sweden)

    Frederico Ozanam Papa

    2012-10-01

    Full Text Available Sperm cryopreservation success depends upon the maintenance of spermatozoa fertility potential. Sperm cells must preserve both integrity and functionality of several cell structures. The stabilization phase must allow the exit of water from the sperm cells via osmosis. This study aimed to compare the effect of refrigeration in the commercial refrigerator (CR and the transport/refrigeration box (TRB upon the viability of frozen bull sperm diluted in three different extenders (A, B and C. Ten Nellore bulls, Bos taurus indicus maintained in Artificial Insemination Center were used and the spermatozoa samples was assessed for Plasma Membrane Integrity and CASA evaluation. The stabilization phase (5°C/4 hours was performed in the CR as well as in the TRB, and then samples were exposed to nitrogen vapor during 20 minutes and then plunged into nitrogen. The statistical analysis was done using the variance analysis and the significance level was set at 5%. In the CR the post-thawing parameters for PM and ALH were higher (p O sucesso da criopreservação do sêmen depende da manutenção do potencial de fertilidade dos espermatozoides. Nos espermatozoides deve haver a preservação da integridade e funcionalidade de várias das suas estruturas. A fase de estabilização permite a saída de água dos espermatozoides por osmose. Este estudo tem o objetivo de comparar o efeito da refrigeração em refrigerador comercial (RC e em caixa de transporte refrigerada (CTR na viabilidade do sêmen congelado bovino diluído em três diferentes meios (A, B e C. Dez touros Nelore, Bos taurus indicus mantidos em central de inseminação artificial foram utilizados e as amostras de sêmen analisadas para checar a integridade das membranas plasmáticas e por meio da análise computadorizada (CASA. A fase de estabilização (5°C/4 horas foi realizada em RC e em CTR, sendo as amostras expostas ao vapor de nitrogênio durante 20 minutos e após mergulhadas no nitrog

  13. Frozen yogurt with added inulin and isomalt.

    Science.gov (United States)

    Isik, U; Boyacioglu, D; Capanoglu, E; Erdil, D Nilufer

    2011-04-01

    The objective of this study was to produce a frozen yogurt containing low fat and no added sugar. Samples containing 5% polydextrose, 0.065% aspartame and acesulfame-K mixture, and different levels of inulin and isomalt (5.0, 6.5, and 8.0%) were produced at pilot scale and analyzed for their physical and chemical properties including proximate composition, viscosity, acidity, overrun, melting rate, heat shock stability, as well as sensory characteristics, and viability of lactic acid bacteria. With the addition of inulin and isomalt, viscosity increased by 19 to 52% compared with that of sample B (reduced-fat control). The average calorie values of samples substituted with sweeteners were about 43% lower than that of original sample. Low-calorie frozen yogurt samples melted about 33 to 48% slower than the reduced-fat control sample at 45 min. Based on quantitative descriptive profile test results, statistically significant differences among products were observed for hardness, iciness, foamy melting, whey separation, and sweetness characteristics. The results of principal component analysis showed that the sensory properties of the sample containing 6.5% inulin and 6.5% isomalt were similar to those of control. Lactic acid bacteria counts of frozen yogurt were found to be between 8.12 and 8.49 log values, 3 mo after the production. The overall results showed that it is possible to produce an attractive frozen yogurt product with the incorporation of inulin and isomalt with no added sugar and reduced fat. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. Enhanced viability and neural differential potential in poor post-thaw hADSCs by agarose multi-well dishes and spheroid culture.

    Science.gov (United States)

    Guo, Xiaoling; Li, Shanyi; Ji, Qingshan; Lian, Ruiling; Chen, Jiansu

    2015-10-01

    Human adipose-derived stem cells (hADSCs) are potential adult stem cells source for cell therapy. But hADSCs with multi-passage or cryopreservation often revealed poor growth performance. The aim of our work was to improve the activity of poor post-thaw hADSCs by simple and effective means. We describe here a simple method based on commercially available silicone micro-wells for creating hADSCs spheroids to improve viability and neural differentiation potential on poor post-thaw hADSCs. The isolated hADSCs positively expresse d CD29, CD44, CD105, and negatively expressed CD34, CD45, HLA-DR by flow cytometry. Meanwhile, they had adipogenic and osteogenic differentiation capacity. The post-thaw and post-spheroid hADSCs from poor growth status hADSCs showed a marked increase in cell proliferation by CKK-8 analysis, cell cycle analysis and Ki67/P27 quantitative polymerase chain reaction (qPCR) analysis. They also displayed an increase viability of anti-apoptosis by annexin v and propidium iodide assays and mitochondrial membrane potential assays. After 3 days of neural induction, the neural differentiation potential of post-thaw and post-spheroid hADSCs could be enhanced by qPCR analysis and western blotting analysis. These results suggested that the spheroid formation could improve the viability and neural differentiation potential of bad growth status hADSCs, which is conducive to ADSCs research and cell therapy.

  15. Comparison of a frozen human foreskin fibroblast cell assay to an enzyme immunoassay and toxigenic culture for the detection of toxigenic Clostridium difficile.

    Science.gov (United States)

    Strachan, Alastair J; Evans, Natalie E; Williams, O Martin; Spencer, Robert C; Greenwood, Rosemary; Probert, Chris J

    2013-01-01

    This study set out to validate the Hs27 ReadyCell assay (RCCNA) as an alternative CCNA method compared against a commonly used commercial enzyme immunoassay (EIA) method and toxigenic culture (TC) reference standard. A total of 860 samples were identified from those submitted to the Health Protection Agency microbiology laboratories over a 30-week period. RCCNA performed much better than EIA when using TC as a gold standard, with sensitivities of 90.8% versus 78.6% and positive predictive value of 87.3% to 81.9%, respectively. The Hs27 Human Foreskin Fibroblast ReadyCells are an easy-to-use and a sensitive CCNA method for the detection of toxigenic Clostridium difficile directly from stool. A turnaround time of up to 48 h for a negative result and possible need for repeat testing make it an unsuitable method to be used in most clinical laboratory setting. Copyright © 2013 Elsevier Inc. All rights reserved.

  16. Comparison of a frozen human foreskin fibroblast cell assay to an enzyme immunoassay and toxigenic culture for the detection of toxigenic Clostridium difficile☆☆☆★

    Science.gov (United States)

    Strachan, Alastair J.; Evans, Natalie E.; Williams, O. Martin; Spencer, Robert C.; Greenwood, Rosemary; Probert, Chris J.

    2013-01-01

    This study set out to validate the Hs27 ReadyCell assay (RCCNA) as an alternative CCNA method compared against a commonly used commercial enzyme immunoassay (EIA) method and toxigenic culture (TC) reference standard. A total of 860 samples were identified from those submitted to the Health Protection Agency microbiology laboratories over a 30-week period. RCCNA performed much better than EIA when using TC as a gold standard, with sensitivities of 90.8% versus 78.6% and positive predictive value of 87.3% to 81.9%, respectively. The Hs27 Human Foreskin Fibroblast ReadyCells are an easy-to-use and a sensitive CCNA method for the detection of toxigenic Clostridium difficile directly from stool. A turnaround time of up to 48 h for a negative result and possible need for repeat testing make it an unsuitable method to be used in most clinical laboratory setting. PMID:23107315

  17. Relation between physical properties of the zona pellucida and viability of bovine embryos after slow-freezing and vitrification.

    Science.gov (United States)

    Moreira da Silva, F; Metelo, R

    2005-06-01

    In vitro-produced bovine morulae/blastocyst embryos (n = 119) were slow-frozen and vitrified and the physical alterations of the zona pellucida (ZP) was observed by scanning electron microscopy (SEM) to find an explanation for the loss of developmental capacity of the embryos after freezing/thawing. A control group was provided, in which embryos (n = 38) were neither frozen nor vitrified. Embryos were in vitro-cultured in a standard CO2 Heraeus incubator and their viability was assessed 24 and 48 h after the start of culture, evaluating their morphological aspect. After 24 h of culture, embryo survival rate for slow-freezing/thawed (n = 23), vitrified/thawed (n = 20) and control embryos (n = 20) was 39, 27 and 90%, and 35, 14 and 65% after 48 h of culture, respectively. For evaluation of physical changes occurring in ZP, 20 embryos were slow-frozen, 18 were vitrified and 18 were used as control. All embryos were fixed, dried and examined under an SEM. Embryo's diameter, as well as the number of pores and their diameter was measured in squares of 6.4 microm width. We observed that, on average, the diameter of the embryos (92.26 +/- 10.15 microm) did not differ significantly among all embryos. As far as the diameter of the pores in the outer surface of the ZP is concerned, the results revealed a significant difference (P embryos. For the number of pores, statistical differences (p embryos (45.4 +/- 7.3 vs 38.2 +/- 8.2). It is possible that ZP functions as a barrier which is positive when dealing with pathogens, but is harmful when nutrients were supplied from the outside, especially at 48 h of culture. Results indicate that the steps of cryopreservation cause alterations in ZP, with irreversible damage on the further developmental competence of bovine embryos.

  18. Effect of inulin and glycerol supplementation on physicochemical properties of probiotic frozen yogurt

    OpenAIRE

    Muzammil, Hafiz Shehzad; Rasco, Barbara; Sablani, Shyam

    2017-01-01

    ABSTRACT The present study was designed to investigate the effects of inulin and glycerol supplementation on physicochemical properties of probiotic frozen yogurt. Frozen yogurt was prepared with different types of probiotic (Lactobacillus acidophilus and Bifidobacterium lactis) along with yogurt starter culture (Streptococcus thermophilus and Lactobacillus bulgaricus). The frozen yogurt mixture was supplemented with inulin (2%, 4%, and 6%) and glycerol (1%, 2%, 3%, and 4%). The results showe...

  19. IMPACT OF PESTICIDES USED IN THE CULTURE OF THE VINE ON THE VIABILITY OF THE YEAST SACCHAROMYCES CEREVISIAE WINE IN CHRONOLOGICAL AGING

    Directory of Open Access Journals (Sweden)

    Alina Owsiak

    2015-02-01

    Full Text Available Pesticides, used in culture, may induce oxidative stress by stimulation of free radicals production, what may result in lipid peroxidation, proteome damage, changes in DNA and RNA structures and disturbance of total antioxidative capacity in organisms’ cells. In disturbances caused by increase synthesis ROS (reactive oxygen species or lack antioxidative defense that is in oxidative stress it is seen one of all causes of aging process. Chronological aging of baker’s and wine yeast Saccharomyces cerevisiae in liquid stationary culture is used as model of research on the aging process. As a result of aging changes take place in yeast cells which have physiological, genetic, metabolic and morphological character, what cause their death in consequence. Some scientists treat chronological yeast aging as analogy of fibroblasts aging of multicellular organisms, skeletal muscles or nerve cells. The aim of the experiment was to obtain the answer on question connected with toxicity effect two widely available pesticides in shape of trade preparation, used among other things in culture of grapevine Miedzian 50 WP (Cu 50WP and Siarkol Extra 80 WP (S 80WP on vitality of wine yeast in chronological aging. During research cells of wine yeast Tokay, which are used in production of white wines, and cells of Malaga strain, used in production of white and red wines, were applied. Yeast culture with pesticides supplementation in determined concentrations was conducted through seven days in YPG medium. At that time vitality of yeast cells was determined by the percentage of cells surviving, percentage of dead cells and culture density. Considerable influence on decreasing vitality of yeast cells in the process of aging showed S 80WP, what correlate with the increase of applied concentration in both example of Malaga and Tokay. Obtained results in application of Cu 50WP indicated lower toxicity in culture of both studied strains in comparison to the control. Our

  20. Breakeven point for energetic viability for the culture of Jatropha; Produtividade minima para a viabilidade energetica da cultura do pinhao-manso

    Energy Technology Data Exchange (ETDEWEB)

    Diotto, Adriano V.; Paulino, Janaina; Romanelli, Thiago L.; Flumgnan, Danilton L.; Folegatti, Marcos V. [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil)], E-mail: avdiotto@usp.br

    2012-11-01

    Due to diversity of plants that can be used to produce biofuel, Jatropha is being increasingly studied and some answers have yet to be given by the research to ensure its use as a plant producing energy. The energy balance is a methodology that takes into account all the energy input in the production system, either through direct consumption of fuel for tractors, for example, or indirectly as the added energy to fabricate the tractor Itself. Therefore, we tried to use the energy balance methodology to evaluate the minimum yield necessary to crop the Jatropha to have energetic viability in production, in other words, to have output energy greater than input energy. We evaluated four years of cultivation in a property located in Charqueada-SP. The feasibility breakeven point was 2.36 mg ha{sup -1}. The factors that most influenced the energy input to the system were the crop inputs (64.3%), followed by fuel consumption (39.7%), depreciation of machinery (3.1%) and labor which accounted for only 1.9%. (author)

  1. What Kind of Signaling Maintains Pluripotency and Viability in Human-Induced Pluripotent Stem Cells Cultured on Laminin-511 with Serum-Free Medium?

    Science.gov (United States)

    Nakashima, Yoshiki; Omasa, Takeshi

    2016-01-01

    Xeno-free medium contains no animal-derived components, but is composed of minimal growth factors and is serum free; the medium may be supplemented with insulin, transferrin, and selenium (ITS medium). Serum-free and xeno-free culture of human-induced pluripotent stem cells (hiPSCs) uses a variety of components based on ITS medium and Dulbecco's modified Eagle's medium/Ham's nutrient mixture F12 (DMEM/F12) that contain high levels of iron salt and glucose. Culture of hiPSCs also requires scaffolding materials, such as extracellular matrix, collagen, fibronectin, laminin, proteoglycan, and vitronectin. The scaffolding component laminin-511, which is composed of α5, β1, and γ1 chains, binds to α3β1, α6β1, and α6β4 integrins on the cell membrane to induce activation of the PI3K/AKT- and Ras/MAPK-dependent signaling pathways. In hiPSCs, the interaction of laminin-511/α6β1 integrin with the cell-cell adhesion molecule E-cadherin confers protection against apoptosis through the Ras homolog gene family member A (RhoA)/Rho kinase (ROCK) signaling pathway (the major pathways for cell death) and the proto-oncogene tyrosine-protein kinase Fyn (Fyn)-RhoA-ROCK signaling pathway. The expression levels of α6β1 integrin and E-cadherin on cell membranes are controlled through the activation of insulin receptor/insulin, FGF receptor/FGF2, or activin-like kinase 5 (ALK5)-dependent TGF-β signaling. A combination of growth factors, medium constituents, cell membrane-located E-cadherin, and α6β1 integrin-induced signaling is required for pluripotent cell proliferation and for optimal cell survival on a laminin-511 scaffold. In this review, we discuss and explore the influence of growth factors on the cadherin and integrin signaling pathways in serum-free and xeno-free cultures of hiPSCs during the preparation of products for regenerative medicinal therapies. In addition, we suggest the optimum serum-free medium components for use with laminin-511, a new scaffold for hi

  2. Viability, invariance and applications

    CERN Document Server

    Carja, Ovidiu; Vrabie, Ioan I

    2007-01-01

    The book is an almost self-contained presentation of the most important concepts and results in viability and invariance. The viability of a set K with respect to a given function (or multi-function) F, defined on it, describes the property that, for each initial data in K, the differential equation (or inclusion) driven by that function or multi-function) to have at least one solution. The invariance of a set K with respect to a function (or multi-function) F, defined on a larger set D, is that property which says that each solution of the differential equation (or inclusion) driven by F and issuing in K remains in K, at least for a short time.The book includes the most important necessary and sufficient conditions for viability starting with Nagumo's Viability Theorem for ordinary differential equations with continuous right-hand sides and continuing with the corresponding extensions either to differential inclusions or to semilinear or even fully nonlinear evolution equations, systems and inclusions. In th...

  3. Microheterogeneity in frozen protein solutions.

    Science.gov (United States)

    Twomey, Alan; Kurata, Kosaku; Nagare, Yutaka; Takamatsu, Hiroshi; Aksan, Alptekin

    2015-06-20

    In frozen and lyophilized systems, the biological to be stabilized (e.g. therapeutic protein, biomarker, drug-delivery vesicle) and the cryo-/lyo-protectant should be co-localized for successful stabilization. During freezing and drying, many factors cause physical separation of the biological from the cryo-/lyo-protectant, called microheterogeneity (MH), which may result in poor stabilization efficiency. We have developed a novel technique that utilized confocal Raman microspectroscopy in combination with counter-gradient freezing to evaluate the effect of a wide range of freezing temperatures (-20frozen formulation in only a few experiments. The freezing experiments conducted with a model system (albumin and trehalose) showed the presence of different degrees of MH in the freeze-concentrated liquid (FCL) in all solutions tested. Mainly, albumin tended to accumulate near the ice interface, where it was physically separated from the cryoprotectant. In frozen 10wt% trehalose solutions, heterogeneity in FCL was relatively low at any TF. In frozen 20wt% trehalose solutions, the optimum albumin to trehalose ratio in the FCL can only be ensured if the solution was frozen within a narrow range of temperatures (-16frozen and stable formulations and freezing protocols for biological as MH is presumed to directly impact stability. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Evaluation of pollen viability, stigma receptivity and fertilization ...

    African Journals Online (AJOL)

    To provide theoretical basis for artificial pollination in Lagerstroemia indica L., pollen viability and stigma receptivity were tested and the morphological change of stigma was observed. Pollen viability tested by in vitro culture, stigma receptivity examined by benzidine-H2O2 testing and fruit set estimated by field artificial ...

  5. Evaluation of pollen viability, stigma receptivity and fertilization ...

    African Journals Online (AJOL)

    AJL

    2013-11-13

    Nov 13, 2013 ... To provide theoretical basis for artificial pollination in Lagerstroemia indica L., pollen viability and stigma receptivity were tested and the morphological change of stigma was observed. Pollen viability tested by in vitro culture, stigma receptivity examined by benzidine-H2O2 testing and fruit set estimated.

  6. Studying frozen movement

    Directory of Open Access Journals (Sweden)

    Jeremy White

    2015-12-01

    Full Text Available Review of Spyros Papapetros, On the Animation of the Inorganic: Art, Architecture, and the Extension of Life: Spyros Papapetros examines ideas about simulated movement and inorganic life during and after the turn of the twentieth century. Exploring works of a selection of important art historians as well as artists and architects of the period, the author maintains that the ability to identify with material objects was repressed by modernist culture, and yet found expression stylistically through depictions of inorganic forms. That expression is shown to have continuity with older medieval and renaissance depictions. The book is organized by a narrative that evokes the modes of inquiry documented and critiqued by the content of the book, employing movement as a narrative device, a metaphor, while serving as a subject of inquiry.

  7. Efficient recovery of fluoroquinolone-susceptible and fluoroquinolone-resistant Escherichia coli strains from frozen samples.

    Science.gov (United States)

    Lautenbach, Ebbing; Santana, Evelyn; Lee, Abby; Tolomeo, Pam; Black, Nicole; Babson, Andrew; Perencevich, Eli N; Harris, Anthony D; Smith, Catherine A; Maslow, Joel

    2008-04-01

    We assessed the rate of recovery of fluoroquinolone-resistant and fluoroquinolone-susceptible Escherichia coli isolates from culture of frozen perirectal swab samples compared with the results for culture of the same specimen before freezing. Recovery rates for these 2 classes of E. coli were 91% and 83%, respectively. The majority of distinct strains recovered from the initial sample were also recovered from the frozen sample. The strains that were not recovered were typically present only in low numbers in the initial sample. These findings emphasize the utility of frozen surveillance samples.

  8. The Eccentric Behavior of Nearly Frozen Orbits

    Science.gov (United States)

    Sweetser, Theodore H.; Vincent, Mark A.

    2013-01-01

    Frozen orbits are orbits which have only short-period changes in their mean eccentricity and argument of periapse, so that they basically keep a fixed orientation within their plane of motion. Nearly frozen orbits are those whose eccentricity and argument of periapse have values close to those of a frozen orbit. We call them "nearly" frozen because their eccentricity vector (a vector whose polar coordinates are eccentricity and argument of periapse) will stay within a bounded distance from the frozen orbit eccentricity vector, circulating around it over time. For highly inclined orbits around the Earth, this distance is effectively constant over time. Furthermore, frozen orbit eccentricity values are low enough that these orbits are essentially eccentric (i.e., off center) circles, so that nearly frozen orbits around Earth are bounded above and below by frozen orbits.

  9. Frozen shoulder and risk of cancer

    DEFF Research Database (Denmark)

    Pedersen, Alma B; Horváth-Puhó, Erzsébet; Ehrenstein, Vera

    2017-01-01

    BACKGROUND: Frozen shoulder might be a complication or a presenting symptom of cancer. We examined the risk of a cancer diagnosis after an incident diagnosis of frozen shoulder. METHODS: We used prospectively collected data from Danish registries to identify patients with frozen shoulder during 1...

  10. 7 CFR 58.327 - Frozen cream.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Frozen cream. 58.327 Section 58.327 Agriculture... Material § 58.327 Frozen cream. To produce frozen cream eligible for official certification, the quality of the cream used shall meet the requirements of cream acceptable for the manufacture of U.S. Grade AA or...

  11. Management and fertility of mares bred with frozen semen.

    Science.gov (United States)

    Samper, J C

    2001-12-03

    Semen quality, mare status and mare management during estrus will have the greatest impact on pregnancy rates when breeding mares with frozen semen. If semen quality is not optimal, mare selection and reproductive management are crucial in determining the outcome. In addition to mare selection, client communication is a key factor in a frozen semen program. Old maiden mares and problem mares should be monitored for normal cyclicity and all, except young maidens, should have at least a uterine culture and cytology performed. Mares with positive bacterial cultures and cytologies should be treated at least three consecutive days when in estrus with the proper antibiotic. With frozen semen, timing the ovulation is highly desirable in order to reduce the interval between breeding and ovulation. The use of ovulation inducing agents such as human chorionic gonadotropin (hCG) or the GnRH analogue, deslorelin, are critical components to accurately time the insemination with frozen semen. Most hCG treated mares ovulate 48h post-treatment (12-72h) while most deslorelin (Ovuplant) treated mares ovulate 36-42h post-treatment. However, mares bred more than once during the breeding cycle appear to have a slight but consistent increase in pregnancy rate compared to mares bred only once pre- or post-ovulation. In addition, the "capacitation-like" changes inflicted on the sperm during the process of freezing and thawing appear to be responsible for the shorter longevity of cryopreserved sperm. Therefore, breeding closer to ovulation should increase the fertility for most stallions with frozen semen. Recent evidence would suggest that breeding close to the uterotubal junction increases the sperm numbers in the oviduct increasing the chances of pregnancy. Post-breeding examinations aid in determining ovulation and uterine fluid accumulations so that post-breeding therapies can be instituted if needed. Average pregnancy rates per cycle of mares bred with frozen semen are between 30 and

  12. Effect of inulin on the physicochemical properties, flow behavior and probiotic survival of frozen yogurt.

    Science.gov (United States)

    Rezaei, Rahil; Khomeiri, Morteza; Aalami, Mehran; Kashaninejad, Mahdi

    2014-10-01

    This study investigated the effect of inulin (0, 1 and 2 %), on some physicochemical properties of frozen yogurt, as well as its effect on flow behavior and probiotic survival. The results showed that the addition of inulin improved overrun, viscosity and melting properties significantly (p frozen yogurt with 2 % inulin had the most appealing sensory characteristics. The flow behavior of all samples showed their pseudoplastic nature; power law was the best model to predict their flow behavior. In terms of probiotic survival, the sample with 2 % inulin significantly improved the viability of Lactobacillus acidophilus and Bifidobacterium lactis.

  13. Improvement In Rabbit Corneal Cell Suspension Viability After Freezing With Gingko Biloba Extrakt

    Directory of Open Access Journals (Sweden)

    Murad Aktan

    2007-01-01

    Full Text Available We investigated whether the addition of Gingko Biloba extract (EGb 761 to rabbit corneal epithelial medium before cell freezing improved cell viability after freezing then thawing. After removal of corneas, they were treated with enzymes and the corneal epithelium was prepared as a single cell suspension in freezing media with or without EGb 761. After freezing for two weeks then thawing, a higher cell viability was found in the cornea cell suspensions which had been frozen pretreated with EGb 761 in the media. The improvement with corneal cell viability with EGb 761 pretreatment is postulated to be based on the antioxidant capacity of the plant extract.

  14. Management of the frozen shoulder

    Directory of Open Access Journals (Sweden)

    van de Laar SM

    2014-10-01

    Full Text Available Suzanne Margaretha van de Laar, Peer van der Zwaal Department of Orthopaedic Surgery and Trauma, Medical Center Haaglanden, The Hague, the Netherlands Abstract: Frozen shoulder is a very common condition with a prevalence of 2%–5% in the general population. Decrease in joint volume as a result of fibrosis and hyperplasia of the joint capsule leads to painful and restricted glenohumeral motion. Frozen shoulder is a self-limiting disease with a chronic character, and is mostly treated in a primary care setting. In this review, we set out to address the current evidence-based literature on management of this disabling disease using a PubMed search. Many non-surgical and surgical therapeutic options are described, including supervised neglect, intra-articular corticosteroid injections, physical therapy, manipulation under anesthesia, capsular distension, and arthroscopic capsular release. In the literature, the long-term outcome shows a significant decrease in pain and improvement of shoulder function for all treatment modalities without clear evidence of superiority of one over the other. This possibly indicates that a self-limiting character is the most important factor in the course of the disease. Management of frozen shoulder is primarily conservative. Supervised neglect is combined with analgesia and stretching exercises as the pain subsides. In the early painful phase, intra-articular corticosteroid injections are recommended for pain relief. When the patient has persistent pain and glenohumeral stiffness after adequate conservative treatment, invasive options can be considered, like arthroscopic capsular release, manipulation under anesthesia, or capsular distension. Keywords: frozen shoulder, adhesive capsulitis, treatment, pathophysiology

  15. Monitoring cell growth, viability, and apoptosis.

    Science.gov (United States)

    Butler, Michael; Spearman, Maureen; Braasch, Katrin

    2014-01-01

    The accurate determination of cell growth and viability is pivotal to monitoring a bioprocess. Direct methods to determine the cell growth and/or viability in a bioprocess include microscopic counting, electronic particle counting, image analysis, in situ biomass monitoring, and dieletrophoretic cytometry. These methods work most simply when a fixed volume sample can be taken from a suspension culture. Manual microscopic counting is laborious but affords the advantage of allowing cell viability to be determined if a suitable dye is included. Electronic particle counting is a rapid total cell count method for replicate samples, but some data distortion may occur if the sample has significant cell debris or cell aggregates. Image analysis based on the use of digital camera images acquired through a microscope has advanced rapidly with the availability of several commercially available software packages replacing manual microscopic counting and viability determination. Biomass probes detect cells by their dielectric properties or their internal concentration of NADH and can be used as a continuous monitor of the progress of a culture. While the monitoring of cell growth and viability is an integral part of a bioprocess, the monitoring of apoptosis induction is also becoming more and more important in bioprocess control to increase volumetric productivity by extending bioprocess duration. Different fluorescent assays allow for the detection of apoptotic characteristics in a cell sample.Indirect methods of cell determination involve the chemical analysis of a culture component or a measure of metabolic activity. These methods are most useful when it is difficult to obtain intact cell samples. However, the relationship between these parameters and the cell number may not be linear through the phases of a cell culture. The determination of nucleic acid (DNA) or total protein can be used as an estimate of biomass, while the depletion of glucose from the media can be used

  16. Comparing effects of freezing at -196 °C and -20 °C on the viability of mastitis pathogens

    Directory of Open Access Journals (Sweden)

    Inge-Marie Petzer

    2012-02-01

    Full Text Available The aim of this study was to compare the effects of cryopreservation at approximately -196 °C in liquid nitrogen (N and freezing at approximately -20 °C in a freezer, on the viability and survival of eight different mastitogenic bacteria inoculated in milk. Bacteria were frozen at approximately -20 °C in a freezer and cryopreserved at approximately -196 °C in liquid nitrogen. An effective preservation method was needed for follow-up samples from cows identified in the South African National Milk Recording Scheme (NMRS with somatic cell counts above 250 000 cells/mL milk. The organisation responsible for sample collection of the NMRS milk samples also provides producers with liquid nitrogen for their semen flasks at the collection sites. This existing mode of storage and transport could therefore be utilised.Ten samples of each organism were thawed and cultured bi-weekly until week 18 for both temperature treatments. An additional sampling was performed at week 30 for samples frozen at approximately -20 °C. Freezing and cryopreservation did not impair subsequent isolation of Streptococcus dysgalactiae, Streptococcus uberis, Enterococcus faecalis, Staphylococcus aureus (STH (phage type lytic group III or Sta. aureus (STA (phage typed, other than lytic group III. Survival was indicated by the isolation of bacteria from samples, and viability by the strength of growth of the bacteria isolated. The survival of Streptococcus agalactiae decreased after week 12 and Escherichia coli after week 16 of freezing, but both organisms survived under cryogenic preservation until week 18. Coagulase-negative staphylococci survived until week 18 for both freezing and cryogenic preservation.Both storage methods could thus contribute to the improvement of a pro-active approach towards udder health management in South African dairy herds.

  17. Evaluation of Physicochemical Characteristics of Smoked River Catfish Prepared from Fresh and Frozen Raws

    Directory of Open Access Journals (Sweden)

    Bustari Hasan

    2016-08-01

    Full Text Available The aim of this study was to evaluate physicochemical characteristics of hotsmoked catfish from freshand frozen fish. River catfish samples (Hemibagrus nemurus Valenciennes, 1840, 240-270 gram in weightwere taken from catfish cage culture in Sungai Paku, Riau. A total fish samples (120 fishes were groupedinto 4 batches, each batch consisted of 30 fish (15 fillets and 15 butterfly like cuts. One batch was smokedfresh and the other 3 batches were smoked after being frozen at -18oC for 10, 20 and 30 days respectively.Before smoked, the fish samples were analyzed for proximate composition and water holding capacity; andafter smoked, the fish samples were determined for smoking yield, proximate composition and sensoryquality. Moisture, fat and protein composition of fish was not different between fresh raw and frozen for10, 20 and 30 days (P>0.05, however, water holding capacity was higher for fresh than frozen fish; andthe value decreased as the longer the frozen storage (P<0.05. Smoking yield correlated stronger to waterholding capacity (r=0.59 than to moisture (r=0.01, fat (r=0.16 and protein (r=0.02 composition of theraw. Moisture, fat and protein of smoked fish was lower for smoked fish prepared from frozen fish than thatfor fresh fish (P<0,05; and the values decreased as the longer the frozen storage. Moisture, fat and proteinloss during smoking was higher for smoked fish prepared from frozen fish than that for fresh fish, exceptfor that frozen for 10 days. Overall, sensory values of smoked fish from frozen fish were lower than that forfresh fish (P<0,05; however, flavor and odor values were not different between smoked fish from fish frozenfor 10 days and fresh fish (P>0,05.

  18. Viability of commercial wine yeasts during freezer storage in glycerol-based media.

    Science.gov (United States)

    Sidari, R; Caridi, A

    2009-01-01

    Glycerol-based medium (BM) with and without the addition of 1 g/L ascorbic acid (Asc) and/or 100 mg/L (+/-)-catechin (Cat) was tested for the storage of three commercial wine yeasts at -20 degrees C. The medium supplemented with Asc was also used to store 706 strains to verify the maintenance of the liquid state. A decline in survival throughout the storage period was observed. The media containing Asc maintained viability better than the other three. The BM caused a loss of viability of 7 orders for one strain and of 6 orders for the other two. All three strains exhibited a loss of viability of 4 orders when stored in BM+Asc. Two strains decreased viability by 5 orders while one strain by 4 orders, when stored in BM+Cat. Two strains decreased viability by 6 orders while one strain by 5 orders, when stored in BM+Asc+Cat. Regarding the physical state of the medium tested on 706 yeast strains, three cases were observed: completely liquid (56.5 %), liquid with only the upper part frozen (40.4 %) without involving the yeast biomass settled at the bottom, and completely frozen (3.12 %). It is practicable to prepare a BM that remains liquid at -20 degrees C enhancing yeast viability when Asc is added as cryoprotectant.

  19. Shoulder pain in primary care: frozen shoulder.

    Science.gov (United States)

    Cadogan, Angela; Mohammed, Khalid D

    2016-03-01

    BACKGROUND AND CONTEXT Frozen shoulder is a painful condition that follows a protracted clinical course. We aim to review the management of patients with a diagnosis of frozen shoulder who are referred for specialist orthopaedic evaluation against existing guidelines in primary care. ASSESSMENT OF PROBLEM Referrals and clinical records were reviewed for all patients referred for orthopaedic specialist assessment who received a specialist diagnosis of frozen shoulder. Diagnostic, investigation and management practices from a regional primary health care setting in New Zealand were compared with guideline-recommended management. RESULTS Eighty patients with frozen shoulder were referred for orthopaedic evaluation in the 13 month study period, mostly from general practice. Fifteen patients (19%) were identified as having a frozen shoulder in their medical referral. Most (99%) had received previous imaging. Seven patients (12%) had received guideline recommended treatment. STRATEGIES FOR IMPROVEMENT Education of all clinicians involved in patient management is important to ensure an understanding of the long natural history of frozen shoulder and provide reassurance that outcomes are generally excellent. HealthPathways now include more information regarding diagnosis, imaging and evidence-based management for frozen shoulder. LESSONS Frozen shoulder may be under-diagnosed among patients referred for orthopaedic review. Ultrasound imaging is commonly used and may identify occult and unrelated pathology in this age-group. When managed according to clinical guidelines, patients report significant clinical and functional improvement with most reporting 80% function compared with normal after 1 year. KEYWORDS Adhesive capsulitis; bursitis; injections; practice guideline; primary health care; ultrasound.

  20. 7 CFR 58.349 - Frozen cream.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Frozen cream. 58.349 Section 58.349 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... Products Bearing Usda Official Identification § 58.349 Frozen cream. The flavor shall be sweet, pleasing...

  1. Frozen yogurt from sheep milk

    Directory of Open Access Journals (Sweden)

    Elisangela de Abreu

    Full Text Available ABSTRACT The aim of this work was to develop frozen yogurt formulations from powdered yogurt of sheep milk, through an experimental design of 2², with a triplicate at the central point. The variables studied were emulsifier/stabilizer (0.50%, 0.75%, and 1.00% and powder for cream (2.75%, 3.00% and 3.25%. The parameters evaluated were sensory characteristics, texture, and microbiological counts. The results showed that the formulations had counts of S. aureus and fecal coliforms at 45 °C, lactic acid bacteria and Salmonella sp within the limits established by legislation. Instrumental analysis of texture-related parameters (firmness, cohesiveness, adhesiveness, and consistency of the formulations with different concentrations of emulsifier/stabilizer and cream powder showed no significant differences (p > 0.05. In sensory analysis, Formulations 3 and 4 with lower concentrations of emulsifier/stabilizer scored the highest values, thus indicating good acceptability.

  2. Variation of Pollen Viability and Storability in Asparagus (Asparagus offcinalis L.) Cultivars

    OpenAIRE

    Ozaki, Yukio; Tashiro, Tomoko; Kurahashi, Tomoko; Okubo, Hiroshi

    1999-01-01

    The optimal culture condition for evaluating pollen viability of asparagus was studied. Sucrose was an effective constituent of the media for raising pollen germination rate. The medium containing 30% sucrose, 0.01% borate and 3% agar was found to be optimum for assessing pollen viability. Effects of temperature and light conditions during in vitro culture on pollen germination rate were not recognized in the range of 20-30℃ in this investigation. Varietal difference of pollen viability and s...

  3. Frozen section diagnosis in ophthalmic pathology

    Directory of Open Access Journals (Sweden)

    Biswas Jyotirmay

    1993-01-01

    Full Text Available Frozen section diagnosis is extensively used in various branches of pathology, but its application in ophthalmic pathology was recognised only in the 1970s. We studied 10 sections of ocular and adenexal lesions by frozen section diagnosis, which included orbital lesions (4 cases, lid lesions (3 cases, and intraocular tissue (1 case. The time taken for processing ranged between 10 to 15 minutes. Diagnoses based on frozen section evaluation included lymphoma, mesenchymal chondrosarcoma, solar keratosis, compound naevus, silicone oil globules in cataractous lens, neurofibromatosis, pseudotumour, retinoblastoma, and chronic blepharitis. Although further histopathologic examination correlated well with the frozen section (100% observations, the diagnosis was deferred in the case of naevus and reactive lymphoid hyperplasia. Our study shows that frozen section diagnosis in ophthalmic surgery is quite reliable and is particularly useful in ocular adenexal lesions

  4. Toxigenic penicillia spoiling frozen chicken nuggets

    DEFF Research Database (Denmark)

    Wigmann, Evelin Francine; Saccomori, Fernanda; Bernardi, Angelica Olivier

    2015-01-01

    Frozen chicken nuggets are classified as pre-prepared frozen meals. These products are convenient to consumers as they are easy to prepare and allow for long storage by freezing. Over the years, spoilage of frozen food products caused by fungi has been a continual problem for the food industry...... since mold can develop when frozen foods are allowed to attain temperatures of -10ºC, or above. The growth of fungi on the food surface results in economic losses and represents a hazard to public health due to the possibility of mycotoxin production. The aim of this study was to identify the species...... citreoviridin, roquefortine C, penitrem A, and verrucosidin under standard conditions. Considering the occurrence of fungal spoilage in frozen food and the potential hazard involved, more studies on psychrophilic fungi growth in foods stored at low temperatures are necessary. (C) 2014 Elsevier Ltd. All rights...

  5. Effect of freezing on rabbit cultured chondrocytes

    Directory of Open Access Journals (Sweden)

    R.R Filgueiras

    2011-02-01

    Full Text Available This work evaluated the effect of freezing on chondrocytes maintained in culture, aiming the establishment of a cell bank for future application as heterologous implant. Chondrocytes extracted from joint cartilage of nine healthy New Zealand White rabbits were cultivated and frozen with the cryoprotector 5% dimethylsulfoxide for six months. Phenotypic and scanning electron microscopy analyses were carried out to identify morphological and functional differences between fresh and thawed cells. After enzymatic digestion, a total of 4.8x10(5cells per rabbit were obtained. Fresh chondrocytes showed a high mitotic rate and abundant matrix was present up to 60 days of culture. Loss of phenotypic stability was notable in the thawed chondrocytes, with a low labeling of proteoglycans and weak immunostaining of type II collagen. The present study showed important loss of chondrocyte viability under the freezing conditions. For future in vivo studies of heterologous implant, these results suggests that a high number of cells should be implanted in the host site in order to achieve an adequate number of viable cells. Furthermore, the chondrocytes should be implanted after two weeks of culture, when the highest viability rate is found

  6. Defective Reduction in Frozen Pie Manufacturing Process

    Science.gov (United States)

    Nooted, Oranuch; Tangjitsitcharoen, Somkiat

    2017-06-01

    The frozen pie production has a lot of defects resulting in high production cost. Failure mode and effect analysis (FMEA) technique has been applied to improve the frozen pie process. Pareto chart is also used to determine the major defects of frozen pie. There are 3 main processes that cause the defects which are the 1st freezing to glazing process, the forming process, and the folding process. The Risk Priority Number (RPN) obtained from FMEA is analyzed to reduce the defects. If RPN of each cause exceeds 45, the process will be considered to be improved and selected for the corrective and preventive actions. The results showed that RPN values decreased after the correction. Therefore, the implementation of FMEA technique can help to improve the performance of frozen pie process and reduce the defects approximately 51.9%.

  7. On water vapour transfer inside frozen packs

    National Research Council Canada - National Science Library

    Heiss, R

    1971-01-01

    In the case of various foodstuffs, irreversible water- vapour transporations within the frozen packages can bring about a distinct quality drop caused by damage to the surface of the material as a result of drying out (freezer burn...

  8. Use of frozen section in genitourinary pathology.

    Science.gov (United States)

    Shen, Steven S; Truong, Luan D; Ro, Jae Y; Ayala, Alberto G

    2012-08-01

    Frozen section diagnosis provides critical information for immediate surgical management decision making. Over the last several years, there have been some significant advances in treatment of genitourinary cancer, particularly with regard to surgical techniques. These changes in turn impact the type and frequency of intraoperative frozen section requests. In this review, we describe the main indications and diagnostic challenges of frozen section diagnosis during surgeries of each genitourinary organ system including prostate, kidney, bladder, testis, and penis. The pitfalls and approaches to different diagnostic situations are discussed. It is also stressed that pathologists must not only be familiar with the histological diagnosis, but also understand the limitations of frozen section diagnosis and communicate with urologists during the intraoperative treatment decision making process.

  9. Cryopreservation in situ of cell monolayers on collagen vitrigel membrane culture substrata: ready-to-use preparation of primary hepatocytes and ES cells.

    Science.gov (United States)

    Miyamoto, Yoshitaka; Enosawa, Shin; Takeuchi, Tomoyo; Takezawa, Toshiaki

    2009-01-01

    Cryopreservation is generally performed on cells in suspension. In the case of adherent cells such as hepatocytes, a loss of their ability to attach is a more serious problem than a decreased viability after cryopreservation. We herein report a novel technology of direct in situ cryopreservation of cells cultured on collagen vitrigel membranes, which have excellent mechanical strength and can be easily handled by tweezers even when coated with cultured cells. Rat primary hepatocytes, mitomycin C-treated mouse fibroblasts (feeder cells for ES cells), and mouse ES cells on the feeder cells were cultured on collagen vitrigel membranes for 1 day. The membranes with cells attached were then plucked up from the dish, soaked in cryopreservation medium containing 10% dimethyl sulfoxide, frozen using a controlled-rate freezer, and transferred to liquid nitrogen. The cells cultured on plastic cell culture dishes were also frozen as controls. After storage in liquid nitrogen for periods from 1 week to 3 months, the cryopreserved membranes with the cells still attached were thawed by adding warmed culture medium. Cell viability estimated by morphology and functional staining with calcein showed significant improvement in comparison to cells cryopreserved without the collagen vitrigel membrane. The recoveries of living cells after cryopreservation were 26.7%, 76.2%, and 58.6% for rat hepatocytes, mitomycin C-treated mouse fibroblasts, and mouse ES cells on collagen vitrigel membranes, respectively. In contrast, essentially no cells at all remained on the plastic cell culture dishes after thawing. Because adherent cell storage under these conditions is very convenient, the use of this technique employing collagen vitrigel membranes should be generally applicable to the cryopreservation of adherent cells that are otherwise problematic to store as frozen stocks.

  10. Seismic modeling using the frozen Gaussian approximation

    OpenAIRE

    Yang, Xu; Lu, Jianfeng; Fomel, Sergey

    2013-01-01

    We adopt the frozen Gaussian approximation (FGA) for modeling seismic waves. The method belongs to the category of ray-based beam methods. It decomposes seismic wavefield into a set of Gaussian functions and propagates these Gaussian functions along appropriate ray paths. As opposed to the classic Gaussian-beam method, FGA keeps the Gaussians frozen (at a fixed width) during the propagation process and adjusts their amplitudes to produce an accurate approximation after summation. We perform t...

  11. Effect of sugar and acid on the acceptability of frozen yogurt to a student population.

    Science.gov (United States)

    Guinard, J X; Little, C; Marty, C; Palchak, T R

    1994-05-01

    One hundred and forty-one college students tasted and rated on a nine-point hedonic scale their degree of liking for nine samples of vanilla frozen yogurt varying in sugar and lactic acid. Subjects were also asked to complete a questionnaire about consumption of frozen yogurt and other dairy products. Degree of liking differed significantly among samples, and the samples best liked were those with the lowest acidity, .23 to .29%, independent of sugar concentration. Degree of liking of frozen yogurt failed to correlate with dairy product consumption or hunger at the time of testing. No significant difference existed between male and female students for overall degree of liking of frozen yogurt or overall dairy product intake, yet the questionnaire revealed a significantly higher consumption of frozen yogurt among female students. The results of this study suggest that, for the student population tested, frozen yogurt should combine the sensory properties of ice cream (low acidity) with the nutritional properties of yogurt (low fat, active enzyme culture).

  12. Persistence of avian influenza viruses in various artificially frozen environmental water types.

    Science.gov (United States)

    Shoham, Dany; Jahangir, Alam; Ruenphet, Sakchai; Takehara, Kazuaki

    2012-01-01

    Background. This study investigates the viable persistence of avian influenza viruses (AIVs) in various types of artificially frozen environmental water and evaluates the feasibility of similar occurrence taking place in nature, and allowing for prolonged abiotic virus survival, with subsequent biotic viral recirculation. Methods. Fresh, brackish, and salty water, taken in Japan from aquatic biotopes regularly visited by migratory waterfowl, were seeded with AIVs. We monthly monitored the viability of the seeded viruses in the frozen state at -20°C and -30°C, for 12 months. We also monitored virus viability following repeatedly induced freezing and thawing. Results. The viruses exhibited considerable viable persistence all along that period of time, as well as during freezing-thawing cycles. Appreciable, yet noncrucial variances were observed in relation to some of the parameters examined. Conclusions. As typical waterborne pathogens of numerous northerly aquatic birds, AIVs are innately adapted to both the body temperature of their hosts (40°C to 42°C) and, presumably, to subzero temperatures of frozen lakes (down to -54°C in parts of Siberia) occupied and virus-seeded by subclinically infected birds, prior to freezing. Marked cryostability of AIVs appears to be evident. Preservation in environmental ice has significant ecophylogenetic and epidemiological implications, potentially, and could account for various unexplained phenomena.

  13. 21 CFR 146.126 - Frozen concentrate for colored lemonade.

    Science.gov (United States)

    2010-04-01

    ... Fruit Juices and Beverages § 146.126 Frozen concentrate for colored lemonade. (a) Frozen concentrate for colored lemonade conforms to the definition and standard of identity prescribed for frozen concentrate for... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen concentrate for colored lemonade. 146.126...

  14. 21 CFR 152.126 - Frozen cherry pie.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen cherry pie. 152.126 Section 152.126 Food... HUMAN CONSUMPTION FRUIT PIES Requirements for Specific Standardized Fruit Pies § 152.126 Frozen cherry pie. (a) Identity. (1) Frozen cherry pie (excluding baked and then frozen) is the food prepared by...

  15. Assessment of viability of microorganisms employing fluorescence techniques

    NARCIS (Netherlands)

    Breeuwer, P.

    1996-01-01


    Viability assessment of microorganisms is relevant for a wide variety of applications in industry, including evaluation of inactivation treatments and quality assessment of starter cultures for beer, wine, and yoghurt production.

    Usually, the ability of microbial cells to

  16. Viability of bull semen extended with commercial semen extender ...

    African Journals Online (AJOL)

    Andrea Raseona

    Abstract. The aim of this study was to evaluate the viability of bull spermatozoa diluted with commercial semen extender and two culture media stored at controlled room temperature (24 °C) for 72 hours. Two Nguni bulls were used for semen collection with the aid of an electro-ejaculator. After macroscopic evaluation ...

  17. Viability of bull semen extended with commercial semen extender ...

    African Journals Online (AJOL)

    After macroscopic evaluation, semen was pooled and aliquoted randomly into Triladyl, modified Ham's F10, and TCM-199 culture media, and then stored at 24 °C. Sperm motility parameters, morphology, and viability were analysed with computer aided sperm analysis (CASA) after 0, 24, 48 and 72 hours. The study was ...

  18. Dormancy, activation and viability of Rhizopus oligosporus sporangiospores

    NARCIS (Netherlands)

    Thanh, N.V.; Nout, M.J.R.

    2004-01-01

    Interruption of dormancy to improve viability of Rhizopus oligosporus sporangiospores is crucial for the application of stored starter cultures for fungal (tempe) production. We aimed to assess the extent of dormancy and factors that could result in activation. Whereas heat treatments were

  19. 21 CFR 146.137 - Frozen orange juice.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen orange juice. 146.137 Section 146.137 Food... Beverages § 146.137 Frozen orange juice. (a) Frozen orange juice is orange juice as defined in § 146.135, except that it is frozen. (b) The name of the food is “Frozen orange juice”. Such name may be preceded on...

  20. THE EXTRACTION OF THE CELL CONTENT OF MICRO-ORGANISMS BY PERCUSSION OF THE FROZEN CELLS

    Science.gov (United States)

    Johlin, J. M.; Avery, Roy C.

    1930-01-01

    A method is described for rapidly obtaining fresh extracts of microorganisms by percussion of the frozen cells. Filtrates giving the biuret reaction and yielding heavy precipitates upon the addition of acetic acid were obtained from the washed cells of cultures of yeast, hemolytic streptococcus, pneumococcus, B. coli, B. xerosis, B. dyphtheriae and the bacillus of bovine tuberculosis. PMID:19869774

  1. Effect of post-slaughter frozen storage on protein, lipid, moisture ...

    African Journals Online (AJOL)

    The effect of post-slaughter frozen storage and exposure on proximate composition and microbial development respectively, were studied in two commonly cultured catfish species, Heterobranchus longifilis and Clarias gariepinus. Samples of minced fillets of males and females of both species were analyzed for protein, ...

  2. The viability of perilabyrinthine osteocytes

    DEFF Research Database (Denmark)

    Bloch, Sune Land; Kristensen, Søren Lund; Sørensen, Mads Sølvsten

    2012-01-01

    Bone remodeling is highly inhibited around the inner ear space, most likely by the anti-resorptive action of the inner ear cytokine osteoprotegerin (OPG) entering perilabyrinthine bone through the lacuno-canalicular porosity (LCP). This extracellular signaling pathway depends on the viability...

  3. Quality assessment of a lot of frozen mackerel, respective frozen handcut mackerel fillets (Scomber scombrus)

    NARCIS (Netherlands)

    Bruenner, K.K.

    2003-01-01

    At the request of Van Ameyde Marine in Amsterdam, RIVO BV assessed the quality of a number of blocks of frozen mackerel and a number of blocks of frozen handcut mackerel fillets (Scomber scombrus) by means of sensorial and chemical analysis. The samples were taken from a lot, which was partial

  4. The Effects of Cryopreservation on Different Passages of Fibroblast Cell Culture in Brown Brocket Deer (Mazama gouazoubira).

    Science.gov (United States)

    Magalhães, Lívia C; Bhat, Maajid H; Freitas, Jeferson L S; Melo, Luciana M; Teixeira, Dárcio I A; Pinto, Luiz C A; Câmara, Lília M C; Duarte, José M B; Freitas, Vicente J F

    2017-10-01

    The brown brocket deer Mazama gouazoubira is 1 of the 10 recognized brocket deer of the Neotropical region. Recently, this species has suffered a population decline due to current threats, mainly poaching and habitat loss. Several studies have shown that some endangered species can benefit from interspecies somatic cell nuclear transfer technology through the use of their somatic cells, such as the fibroblasts. Thus, the aim of this study was to verify the viability and the effect of cryopreservation on fibroblasts after several passages. For this purpose, fibroblast cells were cultured until passages 4, 7, and 10 (cultured control groups) and cryopreserved in cryotubes (frozen/warmed groups). The cellular viability, functionality, and percentage of cells undergoing necrosis and apoptosis were evaluated. The survival rates were always higher than 80% irrespective of the tested group, except for passage 10 in the frozen/warmed group. Population doubling time of cultured cells from passage 10 was significantly higher than that of passages 4 and 7, exhibiting low metabolic activity and a higher percentage of cells in initial apoptosis. In conclusion, the M. gouazoubira fibroblast-derived cell line provides an essential resource for further studies regarding reproductive biotechniques and is likely to be useful as an ex situ conservation strategy.

  5. Culture

    OpenAIRE

    Smith, Timothy B.; Melanie M. Domenech Rodríguez; Bernal, Guillermo

    2011-01-01

    This article summarizes the definitions, means, and research of adapting psychotherapy to clients' cultural backgrounds. We begin by reviewing the prevailing definitions of cultural adaptation and providing a clinical example. We present an original meta-analysis of 65 experimental and quasi-experimental studies involving 8,620 participants. The omnibus effect size of d = .46 indicates that treatments specifically adapted for clients of color were moderately more effective with that clientele...

  6. Viability of pollen grains of tetraploid banana

    Directory of Open Access Journals (Sweden)

    Taliane Leila Soares

    2016-01-01

    Full Text Available ABSTRACT Obtaining banana tetraploid cultivars from triploid strains results in total or partial reestablishment of fertility, allowing the occurrence of some fruits with seeds, a feature that is undesirable from a marketing perspective. The objective of this study was to assess the viability of pollen of 12 banana tetraploid hybrids (AAAB by means of in vitro germination and two histochemical tests (acetocarmine and 2,3,5-triphenyltetrazolium chloride. The pollen tube growth was evaluated by germinating grains in three culture media — M1: 0.03% Ca(NO3∙4H2O, 0.02% Mg(SO4∙7H2O, 0.01% KNO3, 0.01% H3BO3 and 15% sucrose; M2: 0.03% Ca(NO3∙4H2O, 0.01% KNO3, 0.01% H3BO3 and 10% sucrose; and M3: 0.015% H3BO3, 0.045% Ca3(PO42 and 25% sucrose. The acetocarmine staining indicated high viability (above 80%, except for the genotypes YB42-17 and Caprichosa, which were 76 and 70%, respectively. However, the in vitro germination rate was lower than 50% for all the genotypes, except for the hybrids YB42-17 (M1 and YB42-47 (M1. The medium M1 provided the greatest germination percentage and pollen tube growth. Among the genotypes assessed, YB42-47 presented the highest germination rate (61.5% and tube length (5.0 mm. On the other hand, the Vitória cultivar had the lowest germination percentage (8.2% in medium M1. Studies of meiosis can shed more light on the differences observed in the evaluated tetraploids, since meiotic irregularities can affect pollen viability.

  7. Response to intravenous allogeneic equine cord-blood-derived mesenchymal stromal cells administered from chilled or frozen state in serum and protein free media

    Directory of Open Access Journals (Sweden)

    Lynn Brandon Williams

    2016-07-01

    Full Text Available Equine Mesenchymal stromal cells (MSC are commonly transported, chilled or frozen, to veterinary clinics. These MSC must remain viable and minimally affected by culture, transport, or injection processes. The safety of two carrier solutions developed for optimal viability and excipient use were evaluated in ponies, with and without allogeneic cord blood-derived (CB MSC. We hypothesized that neither the carrier solutions nor CB-MSC would elicit measurable changes in clinical, hematological, or biochemical parameters. In 9 ponies (study 1 a bolus of HypoThermosol® FRS (HTS-FRS, CryoStor® CS10 (CS10 or saline was injected IV (n=3/treatment. Study 2, following a one week washout period 5x107 pooled allogeneic CB-MSC were administered IV in HTS-FRS following 24h simulated chilled transport. Study 3, following another one week washout period 5x107 pooled allogeneic CB-MSC were administered IV in CS10 immediately after thawing. Nine ponies received CB-MSCs in study 2 and 3 and three ponies received the cell carrier media without cells. CB-MSCs were pooled in equal numbers from five unrelated donors. In all studies ponies were monitored with physical examination, and blood collection for 7 days following injection. CD4 and CD8 lymphocyte populations were also evaluated in each blood sample.In all three studies, physical exam, complete blood cell count, serum biochemistry, and coagulation panel did not deviate from established normal ranges. Proportions of CD4+ and CD8+ lymphocytes increased at 168h post injection in CB-MSC treatment groups regardless of the carrier solution. Decreases in CD4+/CD8+ double positive populations were observed at 24 h and 72 h in CB-MSC treated animals. There was no difference in viability between CB-MSC suspended in HTS-FRS or CS10.HTS-FRS and CS10 used for low volume excipient injection of MSC suspensions was not associated with short-term adverse reactions. HTS-FRS and CS10 both adequately maintain CB-MSC viability

  8. [Transplantation of deep frozen menisci].

    Science.gov (United States)

    Pasa, L; Pokorný, V; Kalandra, S; Melichar, I; Bilik, A

    2008-02-01

    Subtotal or total meniscectomy will increase weight-bearing per square unit of the cartilage surface approximately threeand- half-times. A long-term overloading of cartilage is clinically manifested by pain, swelling and a rapid onset of early arthritic lesions discernible on radiograms. One of the options for the treatment of degenerative changes in the joint is meniscal transplant. The authors present their first experience with the transplantation of deep frozen meniscal tissue in the Czech Republic. By September 2006, we had treated 26 patients with clinical problems following subtotal or total meniscectomy. The patients, 15 women and 11 men, were between 24 and 46 years of age. Eighteen patients underwent transplantation of the medial meniscus and eight received a lateral meniscal transplant. Concomitant repair of the anterior cruciate ligament (ACL) was indicated in 11 patients, of whom 10 were treated with semitendinosus tendon graft and one with patellar ligament allograft. One patient with a lateral meniscal transplant and ACL reconstruction also had suture of the medial meniscus for a previously sustained injury. In 16 patients, chondromalacia was at the level of Outerbridge grade II and, only in five patients, the finding was Outerbridge grade I. Five patients with grade III chondromalacia were treated using the microfracture technique. Valgus or varus osteotomy was not indicated at all. The goal of meniscal transplant surgery is: 1) to relieve pain after meniscectomy; 2) to prevent degenerative changes of cartilage; 3) to eliminate or reduce the risk of development of osteoarthritic lesions; 4) to restore normal mechanics of the knee joint. Patient selection is important and it is necessary to take into consideration: 1) level of cartilage degenerative changes; 2) knee alignment; 3) knee joint stability; 4) graft size. In patients with instability of the knee and indications for meniscal graft, it is necessary to stabilize the joint by ligament

  9. Culture

    OpenAIRE

    Coulangeon, Philippe

    2013-01-01

    Il n’est sans doute pas de notion aussi vaste et aussi polysémique en sciences sociales que la notion de culture, qui renvoie alternativement à l’ensemble des symboles, des significations, des valeurs et des manières de faire propres à un groupe et au domaine spécialisé des activités expressives, savantes et populaires. La notion de culture est ainsi tout autant mobilisée dans l’exploration des grandes thématiques de la sociologie (stratification, inégalités, institutions, mouvements sociaux)...

  10. Effect of B-mercaptoethanol on the viability of IVM/IVF/IVC bovine embryos during long-distance transportation in plastic straws.

    Science.gov (United States)

    Takahashi, H; Kuwayama, M; Hamano, S; Takahashi, M; Okano, A; Kadokawa, H; Kariya, T; Nagai, T

    1996-10-15

    Experiments were conducted to assess the effect of beta-mercaptoethanol (beta-ME) on the quality and viability of bovine blastocysts derived from in-vitro culture (IVC) of in-vitro matured and fertilized (TVM-IVF) oocytes during their transport between 2 distant places. Follicular oocytes were collected from ovaries obtained at a slaughterhouse and were cultured for 20 to 21 h in modified TCM-199. The IVM oocytes were fertilized in vitro with frozen-thawed spermatozoa. Fertilized oocytes were cultured for 7 d, and embryos that developed to the blastocyst stage were used for the experiments. The blastocysts, packed in straws with transportation medium that consisted of modified TCM-199 with HEPES equilibrated in air and supplemented with 20 % calf serum and 0, 10, 50, 100 or 150 microM beta-ME, were transported at 37 degrees C from Tokyo to Sapporo by air (18.3 h). The quality of blastocysts was assessed and ranked as excellent (A), good (B), fair (C) or poor (D) after transportation. The percentages of blastocysts ranked as A or B were significantly higher (P plastic straws for several hours without control of CO2 and that the concentration of beta-ME used in this experiment is not detrimental to the blastocysts.

  11. Cultural

    Science.gov (United States)

    Wilbur F. LaPage

    1971-01-01

    A critical look at outdoor recreation research and some underlying premises. The author focuses on the concept of culture as communication and how it influences our perception of problems and our search for solutions. Both outdoor recreation and science are viewed as subcultures that have their own bodies of mythology, making recreation problems more difficult to...

  12. Assessment of strategies to increase chondrocyte viability in cryopreserved human osteochondral allografts: evaluation of the glycosylated hydroquinone, arbutin.

    Science.gov (United States)

    Rosa, S C; Gonçalves, J; Judas, F; Lopes, C; Mendes, A F

    2009-12-01

    Allogeneic cartilage is used to repair damaged areas of articular cartilage, requiring the presence of living chondrocytes. So far, no preservation method can effectively meet that purpose. Identification of more effective cryoprotective agents (CPAs) can contribute to this goal. The aim of this study was to determine whether the glycosylated hydroquinone, arbutin, alone or in combination with low concentrations of other CPAs, has cryoprotective properties towards human articular cartilage. Human tibial plateaus were procured from multi-organ donors, with the approval of the Ethics Committee of the University Hospital of Coimbra. The tibial plateaus were treated with or without arbutin (50 or 100mM), alone or in combination with various concentrations of dimethyl sulfoxide (DMSO) and glycerol, for 0.5-1.5h/37 degrees C, then frozen at -20 degrees C and 24h later transferred to a biofreezer at -80 degrees C. Two to 3 months later, thawing was achieved by immersion in cell culture medium at 37 degrees C/1h. Chondrocyte viability was assessed before and after freeze-thawing using a colorimetric assay based on the cell's metabolic activity and fluorescent dyes to evaluate cell membrane integrity. Before freezing, chondrocyte metabolic activity was identical in all the conditions tested. After freeze-thawing, the highest activity, corresponding to 34.2+/-2.1% of that in the Fresh Control, was achieved in tibial plateaus incubated in 50mM arbutin for 1h whereas in those left untreated it was 11.1+/-4.7. Addition of DMSO and glycerol to arbutin did not increase chondrocyte viability any further. Fluorescence microscopy confirmed these results and showed that living chondrocytes were mainly restricted to the superficial cartilage layers. Arbutin seems to be an effective cryoprotective agent for osteochondral allografts with potential benefits over DMSO and glycerol.

  13. Frozen: The Potential and Pitfalls of Ground-Penetrating Radar for Archaeology in the Alaskan Arctic

    Directory of Open Access Journals (Sweden)

    Thomas M. Urban

    2016-12-01

    Full Text Available Ground-penetrating radar (GPR offers many advantages for assessing archaeological potential in frozen and partially frozen contexts in high latitude and alpine regions. These settings pose several challenges for GPR, including extreme velocity changes at the interface of frozen and active layers, cryogenic patterns resulting in anomalies that can easily be mistaken for cultural features, and the difficulty in accessing sites and deploying equipment in remote settings. In this study we discuss some of these challenges while highlighting the potential for this method by describing recent successful investigations with GPR in the region. We draw on cases from Bering Land Bridge National Preserve, Cape Krusenstern National Monument, Kobuk Valley National Park, and Gates of the Arctic National Park and Preserve. The sites required small aircraft accessibility with light equipment loads and minimal personnel. The substrates we investigate include coastal saturated active layer over permafrost, interior well-drained active layer over permafrost, a frozen thermo-karst lake, and an alpine ice patch. These examples demonstrate that GPR is effective at mapping semi-subterranean house remains in several contexts, including houses with no surface manifestation. GPR is also shown to be effective at mapping anomalies from the skeletal remains of a late Pleistocene mammoth frozen in ice. The potential for using GPR in ice and snow patch archaeology, an area of increasing interest with global environmental change exposing new material each year, is also demonstrated.

  14. Influence of freezing temperatures prior to freeze-drying on viability of yeasts and lactic acid bacteria isolated from wine.

    Science.gov (United States)

    Polo, L; Mañes-Lázaro, R; Olmeda, I; Cruz-Pio, L E; Medina, Á; Ferrer, S; Pardo, I

    2017-06-01

    To determine the effect of three different freezing temperatures on post-freeze-drying survival rates of wine yeasts and lactic acid bacteria (LAB). To know if a similar freeze-drying protocol can be used for both micro-organisms. Cells from liquid culture media were recovered and concentrated in appropriate lyoprotectants. Aliquots of each strain were frozen at -20, -80 and -196°C before vacuum drying. Viable cell counts were done before freezing and after freeze-drying. Survival rates were calculated. Freezing temperatures differently affected yeast and bacteria survival. The highest survival rates were obtained at -20 and -80°C for yeasts, but at -196°C for LAB. Major differences in survival rates were recorded among freeze-dried yeasts, but were less drastic for LAB. Yeasts Pichia membranifaciens, Starmerella bacillaris and Metschnikowia pulcherrima, and LAB Lactobacillus paracasei, Pediococcus parvulus and Lactobacillus mali, were the most tolerant species to freeze-drying, regardless of freezing temperature. Yeast and LAB survival rates differed for each tested freezing temperature. For yeasts, -20°C ensured the highest post-freeze-drying viability and -196°C for LAB. Freezing temperature to freeze-dry cells is a crucial factor for ensuring good wine yeast and LAB survival. These results are important for appropriately preserving micro-organisms and for improving starter production processes. © 2017 The Society for Applied Microbiology.

  15. Effect of starter culture and inulin addition on microbial viability, texture, and chemical characteristics of whole or skim milk Kefir Efeito do tipo de cultura starter e da adição de inulina na viabilidade microbiana, textura e características químicas de Kefir de leite integral ou desnatado

    Directory of Open Access Journals (Sweden)

    Flávia Daiana Montanuci

    2012-12-01

    Full Text Available The effect of inulin addition and starters (Kefir grains or commercial starter culture on the microbial viability, texture, and chemical characteristics of Kefir beverages prepared with whole or skim milk was evaluated during refrigerated storage. The type of starter did not influence microbial viability during the storage of the beverages, but the chemical and textural changes (decreases in pH, lactose concentration, and inulin and increased acidity, firmness, and syneresis were more pronounced in the formulations fermented with grains than those fermented with the starter culture. The addition of inulin did not influence acidity or viability of lactic acid bacteria, but in general, its effect on the survival of acetic acid bacteria, Lactococcus and yeasts, firmness, and syneresis depended on the type of milk and starter culture used. Generally, the yeast, acetic acid bacteria, and Leuconostoc counts increased or remained unchanged, while the total population of lactic acid bacteria and Lactococcus were either reduced by 1 to 2 logs or remained unchanged during storage.O efeito da adição de inulina e do tipo de iniciador (grãos de Kefir ou cultura starter comercial da fermentação sobre a viabilidade microbiana, textura e características químicas de bebidas Kefir, formuladas com leite integral ou desnatado, foi avaliado durante o armazenamento refrigerado. O tipo de iniciador não teve influência sobre a viabilidade microbiana ao longo da estocagem das bebidas, mas as alterações químicas e de textura (redução do pH, teores de lactose e inulina e aumento da acidez, firmeza e sinérese foram mais acentuadas nas formulações fermentadas com grãos do que com cultura starter. A adição de inulina não influenciou a acidez ou a viabilidade de bactérias ácido-láticas, mas, em geral, seu efeito sobre a sobrevivência das bactérias ácido-acéticas, Lactococcus e leveduras, firmeza e sinérese foi dependente do tipo de leite e da

  16. Recent developments in the use of viability dyes and quantitative PCR in the food microbiology field

    National Research Council Canada - National Science Library

    Elizaquível, P; Aznar, R; Sánchez, G

    2014-01-01

    .... Moreover, those dyes have been explored to monitor different food manufacturing processes as an alternative to classical cultural methods. In this review, state‐of‐the‐art information regarding viability PCR (v‐PCR) is compiled.

  17. Production of bovine cloned embryos with donor cells frozen at a slow cooling rate in a conventional freezer (20 C)

    Science.gov (United States)

    Chacon, L.; Gomez, M.C.; Jenkins, J.A.; Leibo, S.P.; Wirtu, G.; Dresser, B.L.; Pope, C.E.

    2009-01-01

    Summary Usually, fibroblasts are frozen in dimethyl sulphoxide (DMSO, 10% v/v) at a cooling rate of 1 C/min in a low-temperature (80 C) freezer (LTF) before storage in liquid nitrogen (LN2); however, a LTF is not always available. The purpose of the present study was to evaluate apoptosis and viability of bovine fibroblasts frozen in a LTF or conventional freezer (CF; 20 C) and their subsequent ability for development to blastocyst stage after fusion with enucleated bovine oocytes. Percentages of live cells frozen in LTF (49.5%) and CF (50.6%) were similar, but significantly less than non-frozen control (88%). In both CF and LTF, percentages of live apoptotic cells exposed to LN2 after freezing were lower (4% and 5%, respectively) as compared with unexposed cells (10% and 18%, respectively). Cells frozen in a CF had fewer cell doublings/24 h (0.45) and required more days (9.1) to reach 100% confluence at the first passage (P) after thawing and plating as compared with cells frozen in a LTF (0.96 and 4.0 days, respectively). Hypoploidy at P12 was higher than at P4 in cells frozen in either a CF (37.5% vs. 19.2%) or in a LTF (30.0% vs. 15.4%). A second-generation cryo-solution reduced the incidence of necrosis (29.4%) at 0 h after thawing as compared with that of a first generation cryo-solution (DMEM + DMSO, 60.2%). The percentage of apoptosis in live cells was affected by cooling rate (CF = 1.9% vs. LFT = 0.7%). Development of bovine cloned embryos to the blastocyst stage was not affected by cooling rate or freezer type. ?? 2009 Cambridge University Press.

  18. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  19. Measuring thrombin activity in frozen brain tissue.

    Science.gov (United States)

    Reuveni, Gilad; Golderman, Valery; Shavit-Stein, Efrat; Rosman, Yossi; Shrot, Shai; Chapman, Joab; Harnof, Sagi

    2017-12-06

    Thrombin is a coagulation factor implicated in various pathological and physiological processes in the brain, exerting beneficial and deleterious effects in a concentration-dependent manner. Measurement of thrombin activity levels in pathological animal models is needed and in some cases, because of technical considerations, only frozen samples are available. In the current study, we used a quantitative method to evaluate thrombin activity in fresh and frozen brain sections of 43 male and female adult healthy mice. We stratified data per brain section, brain hemisphere, and mouse sex. We found lower thrombin activity in frozen sections compared with fresh sections, falling within levels considered central nervous system protective in previous studies. The results suggest that fresh section thrombin activity levels in healthy mice can be extrapolated from frozen brain sections. In addition, we found varying thrombin activity across the brain sections, with maximal activity in the olfactory system and hippocampus-containing sections. Thrombin activity did not vary between males and females, or between the right and the left hemispheres, in a statistically significantly manner.

  20. Resurrection of a bull by cloning from organs frozen without cryoprotectant in a -80 degrees c freezer for a decade.

    Science.gov (United States)

    Hoshino, Yoichiro; Hayashi, Noboru; Taniguchi, Shunji; Kobayashi, Naohiko; Sakai, Kenji; Otani, Tsuyoshi; Iritani, Akira; Saeki, Kazuhiro

    2009-01-01

    Frozen animal tissues without cryoprotectant have been thought to be inappropriate for use as a nuclear donor for somatic cell nuclear transfer (SCNT). We report the cloning of a bull using cells retrieved from testicles that had been taken from a dead animal and frozen without cryoprotectant in a -80 degrees C freezer for 10 years. We obtained live cells from defrosted pieces of the spermatic cords of frozen testicles. The cells proliferated actively in culture and were apparently normal. We transferred 16 SCNT embryos from these cells into 16 synchronized recipient animals. We obtained five pregnancies and four cloned calves developed to term. Our results indicate that complete genome sets are maintained in mammalian organs even after long-term frozen-storage without cryoprotectant, and that live clones can be produced from the recovered cells.

  1. Theoretical study of thermal damage in frozen soil

    Directory of Open Access Journals (Sweden)

    Zhu Zhi-Wu

    2015-01-01

    Full Text Available The weakened strength of frozen soil caused by rising temperature can result in thermal damage, which is mainly affected by ice content. A dynamic model for frozen ice is proposed considering temperature and volumetric strain.

  2. Effects of ice-seeding temperature and intracellular trehalose contents on survival of frozen Saccharomyces cerevisiae cells.

    Science.gov (United States)

    Nakamura, Toshihide; Takagi, Hiroshi; Shima, Jun

    2009-04-01

    Freezing tolerance is an important characteristic for baker's yeast, Saccharomyces cerevisiae, as it is used to make frozen dough. The ability of yeast cells to survive freezing is thought to depend on various factors. The purpose of this work was to study the viability of yeast cells during the freezing process. We examined factors potentially affecting their survival, including the growth phase, ice-seeding temperature, intracellular trehalose content, freezing period, and duration of supercooling. The results showed that the ice-seeding temperature significantly affected cell viability. In the stationary phase, trehalose accumulation did not affect the viability of yeast cells after brief freezing, although it did significantly affect the viability after prolonged freezing. In the log phase, the ice-seeding temperature was more important for cell survival than the presence of trehalose during prolonged freezing. The importance of increasing the extracellular ice-seeding temperature was verified by comparing frozen yeast survival rates in a freezing test with ice-seeding temperatures of -5 degrees C and -15 degrees C. We also found that the cell survival rates began to increase at 3h of supercooling. The yeast cells may adapt to subzero temperatures and/or acquire tolerance to freezing stress during the supercooling.

  3. Culture

    OpenAIRE

    Dubois, Vincent

    2007-01-01

    Le découpage des spécialités sociologiques hésite habituellement entre une répartition thématique par domaines empiriquement distingués et un partage conceptuel reposant sur des orientations de recherche. La sociologie de la culture n'échappe pas à cette oscillation. De prime abord, elle couvre un secteur plus ou moins clairement délimité, qui englobe la sociologie de l'art et ce qui est socialement désigné comme relevant de la « vie culturelle ». Elle regroupe alors un ensemble de subdivisio...

  4. Tychastic measure of viability risk

    CERN Document Server

    Aubin, Jean-Pierre; Dordan, Olivier

    2014-01-01

    This book presents a forecasting mechanism of the price intervals for deriving the SCR (solvency capital requirement) eradicating the risk during the exercise period on one hand, and measuring the risk by computing the hedging exit time function associating with smaller investments the date until which the value of the portfolio hedges the liabilities on the other. This information, summarized under the term “tychastic viability measure of risk” is an evolutionary alternative to statistical measures, when dealing with evolutions under uncertainty. The book is written by experts in the field and the target audience primarily comprises research experts and practitioners.

  5. Use of fresh versus frozen or blast-frozen grapes for small-scale fermentation

    Directory of Open Access Journals (Sweden)

    Schmid F

    2011-10-01

    Full Text Available Frank Schmid, Vladimir Jiranek School of Agriculture, Food and Wine, The University of Adelaide; and Wine Innovation Cluster, The Waite Campus, Glen Osmond, South Australia, Australia Background: This paper firstly examines the validity of using laboratory-scale fermentations as a means of correlating winemaking outcomes with larger industrial scale fermentations. Secondly, conventional and blast-freezing of whole bunches were investigated for their relative suitability as methods of preservation as determined by the nature of the resulting wines. Methods: Red must fermentations were compared at the laboratory 80 kg scale, and the more industrially representative 500 kg pilot scale. Fermentation profiles and duration for both scales were found to be very similar. Whole bunches were either slow/conventionally frozen (−20°C, or quickly/blast-frozen (−25°C. Results: Wines made from frozen grapes compared well with the wine made from the fresh must. Color and chemical analyses of the wines revealed few differences. A duo-trio sensory evaluation showed that wine from blast-frozen grapes was more similar to the fresh wines than wines from conventional frozen grapes. Conclusion: The findings of this research suggest that whole-bunch blast-freezing of grapes is preferable to conventional freezing. Keywords: wine color, research winemaking, frozen grapes

  6. Comparative analysis of wholesale and retail frozen fish marketing ...

    African Journals Online (AJOL)

    Therefore, null hypothesis was rejected. High costs of transportation, inadequate supply of frozen fish in the market were the major constraints in the frozen fish marketing. It is recommended that government should put in place policies that would enhance frozen fish marketers' access to loans in order to increase their gross ...

  7. 48 CFR 852.246-72 - Frozen processed foods.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Frozen processed foods. 852.246-72 Section 852.246-72 Federal Acquisition Regulations System DEPARTMENT OF VETERANS AFFAIRS... Frozen processed foods. As prescribed in 846.302-72, insert the following clause: Frozen Processed Foods...

  8. 48 CFR 870.111-5 - Frozen processed food products.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Frozen processed food... DEPARTMENT SUPPLEMENTARY REGULATIONS SPECIAL PROCUREMENT CONTROLS Controls 870.111-5 Frozen processed food products. (a) The following frozen processed food products must have a label complying with the Federal...

  9. 48 CFR 846.302-72 - Frozen processed foods.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Frozen processed foods... CONTRACT MANAGEMENT QUALITY ASSURANCE Contract Clauses 846.302-72 Frozen processed foods. The contracting officer shall insert the clause at 852.246-72, Frozen processed foods, in solicitations and contracts for...

  10. 21 CFR 864.9145 - Processing system for frozen blood.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Processing system for frozen blood. 864.9145... Blood and Blood Products § 864.9145 Processing system for frozen blood. (a) Identification. A processing system for frozen blood is a device used to glycerolize red blood cells prior to freezing to minimize...

  11. Batch testing for noroviruses in frozen raspberries.

    Science.gov (United States)

    De Keuckelaere, Ann; Li, Dan; Deliens, Bart; Stals, Ambroos; Uyttendaele, Mieke

    2015-01-02

    Berries, in particular raspberries, have been associated with multiple recalls due to norovirus contamination and were linked to a number of norovirus (NoV) outbreaks. In the present study a total of 130 samples of frozen raspberries were collected from 26 batches in four different raspberry processing companies. In two companies the samples consisted of bulk frozen raspberries serving as raw material for the production of raspberry puree (an intermediate food product in a business to business setting). In two other companies, the samples consisted of bulk individually quick frozen (IQF) raspberries serving as raw material for the production of frozen fruit mixes (as a final food product for consumer). Enumeration of Escherichia coli and coliforms was performed as well as real-time reverse transcription PCR (RT-qPCR) detection of GI and GII NoV (in 2 × 10 g). In addition, in cases where positive NoV GI or GII RT-qPCR signals were obtained, an attempt to sequence the amplicons was undertaken. Six out of 70 samples taken from the 14 batches of frozen raspberries serving raspberry puree production provided a NoV RT-qPCR signal confirmed by sequencing. Four of these six positive samples clustered in one batch whereas the other two positive samples clustered in another batch from the same company. All six positive samples showed NoV RT-qPCR signals above the limit of quantification of the RT-qPCR assay. These two positive batches of frozen raspberries can be classified as being of insufficient sanitary quality. The mean NoV level in 20 g of these raspberry samples was 4.3 log genomic copies NoV GI/20 g. The concern for public health is uncertain as NoV RT-qPCR detection is unable to discriminate between infectious and non-infectious virus particles. For the IQF raspberries, one batch out of 12 tested NoV positive, but only 1 out of the 5 samples analyzed in this batch showed a positive RT-qPCR GI NoV signal confirmed by sequencing. The RT-qPCR signal was below the

  12. Culture

    OpenAIRE

    Boas, Franz

    2003-01-01

    L’un des objets de l’enquête anthropologique, pour laquelle des éléments peuvent être obtenus par l’étude des sociétés existantes, est l’inter-dépendance des phénomènes culturels. Alors que dans l’étude des processus de diffusion et de développement parallèle les caractères et la distribution de traits singuliers sont communément les objets de l’analyse, nous sommes conduits, ici, à considérer la culture, dans toutes ses manifestations, comme un tout. Les inventions, la vie économique, la str...

  13. Toxicity potential of residual ethylene oxide on fresh or frozen embryos maintained in plastic straws.

    Science.gov (United States)

    Schiewe, M C; Schmidt, P M; Pontbriand, D; Wildt, D E

    1988-01-01

    The toxic effects of residual ethylene oxide (EtO), a frequently used gas-sterilant, on embryos either frozen for long-term purposes or stored acutely for 30 min to 9 hr in a fresh condition in 0.25-ml straw containers were evaluated. In Experiment 1, fresh embryos were frozen (using conventional technology) in straws previously aerated for 0 hr to 8 mo after EtO sterilization. With the exception of the 8-mo group in which survival and quality ratings were depressed, embryo viability was not affected significantly by short-term prefreeze and post-thaw exposure to EtO residues. Experiment 2 was conducted to analyze the influence of prefreeze exposure to EtO residues on embryo development in vitro for embryos temporarily stored in previously sterilized straws aerated for different intervals. Compared to non-EtO-sterilized control straws, the development, quality, and viability of embryos exposed to EtO-treated straws were compromised (p less than 0.05) as the aeration interval decreased and the exposure interval increased. The combined results of both experiments indicate that EtO-treated straws can be used to cryopreserve gametes efficiently, but only if the aeration interval is greater than or equal to 72 hr and the prefreeze duration of exposure is less than or equal to 3 hr.

  14. Frozen shoulder and the Big Five personality traits.

    Science.gov (United States)

    Debeer, Philippe; Franssens, Fien; Roosen, Isabelle; Dankaerts, Wim; Claes, Laurence

    2014-02-01

    In the past, several studies have suggested the existence of a "periarthritic personality" in patients with frozen shoulder. We conducted a study to determine differences in personality traits in patients with primary and secondary frozen shoulders. We prospectively evaluated 118 patients (84 women and 34 men; mean age, 53.8 years; SD 7.56) with a frozen shoulder. Of these patients, 48 had an idiopathic frozen shoulder and 70 had a secondary frozen shoulder. Personality traits were determined by the NEO Five-Factor Inventory (NEO-FFI) scale. This questionnaire measures the 5 major personality traits and is based on the norms determined in a neutral test situation for 2415 controls. Compared with healthy controls, no differences in personality traits were found in patients with primary and secondary frozen shoulder, except for Conscientiousness and Extraversion, for which patients with secondary frozen shoulder scored significantly higher than healthy controls. Patients with primary frozen shoulder scored significantly higher on Openness to Experience than did patients with secondary frozen shoulder; on the other 4 Big Five personality traits, no significant differences were found between patients with primary and secondary frozen shoulder. More specifically, patients with idiopathic frozen shoulder did not score higher on the trait Neuroticism as would be expected from previous publications. Our study results do not indicate that patients with an idiopathic frozen shoulder have a specific personality compared with healthy controls. Only a few differences were found in personality traits when the entire frozen shoulder group was compared with healthy controls and between patients with primary and secondary frozen shoulders. The results of this study suggest that these differences are not sufficient to speak about a specific "frozen shoulder personality." Copyright © 2014 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Mosby, Inc. All rights

  15. IN VITRO CULTURE OF FROZEN AND THAWED MOUSE OVA

    OpenAIRE

    KANAGAWA, Hiroshi

    1980-01-01

    Seventy mouse ova at the 4--8-cell stages were collected at room temperature from mice treated with pregnant mare's serum gonadotrophin and human chorionic gonadotrophin. Seventy ova were divided into 5 groups, and each group was placed in a 0.5 ml plastic straw with 0.2 ml of Brinster's medium. Then, the straws were immersed into an ice bath (0℃) for 5 minutes. Next, an equal volume of 2 M dimethyl sulfoxide was added to the sample straw. The medium with ova was then seeded at -6℃ and cooled...

  16. The Jefferson Lab Frozen Spin Target

    Energy Technology Data Exchange (ETDEWEB)

    Christopher Keith, James Brock, Christopher Carlin, Sara Comer, David Kashy, Josephine McAndrew, David Meekins, Eugene Pasyuk, Joshua Pierce, Mikell Seely

    2012-08-01

    A frozen spin polarized target, constructed at Jefferson Lab for use inside a large acceptance spectrometer, is described. The target has been utilized for photoproduction measurements with polarized tagged photons of both longitudinal and circular polarization. Protons in TEMPO-doped butanol were dynamically polarized to approximately 90% outside the spectrometer at 5 T and 200-300 mK. Photoproduction data were acquired with the target inside the spectrometer at a frozen-spin temperature of approximately 30 mK with the polarization maintained by a thin, superconducting coil installed inside the target cryostat. A 0.56 T solenoid was used for longitudinal target polarization and a 0.50 T dipole for transverse polarization. Spin relaxation times as high as 4000 hours were observed. We also report polarization results for deuterated propanediol doped with the trityl radical OX063.

  17. Microwave thawing of frozen parenteral solutions.

    Science.gov (United States)

    Walter, C W; Pauly, J A; Ausman, R K; Kundsin, R B; Holmes, C J

    1983-01-01

    A commercially available microwave oven modified for use at medication stations throughout hospitals allows timely thawing of frozen parenteral solutions. The inherent problems of safety and uniform heating have been overcome, thus making possible the preparation, storage, and distribution of admixtures on a regional basis and ensuring the integrity of the product. Most parenteral medications are not degraded by microwave energy, and thawing by microwave energy permits timely administration and allows coordination of medication for a series of patients.

  18. Safety of frozen liver for human consumption

    Directory of Open Access Journals (Sweden)

    Ghada A.K. Kirrella

    2017-07-01

    Full Text Available The objective of this study was to ensure and evaluate the safety of imported frozen beef liver traded in supermarkets of Kafr El-Sheikh Governorate, Egypt, through detection of Salmonella typhimurium, Salmonella enteritidies, Escherichia coli O157:H7, antibiotic residues, and aflatoxin B1 residue. Fifty samples of imported frozen liver were randomly collected from different shops at Kafr El-Sheikh Governorate for isolation of S. typhimurium, S. enteritidies, and E. coli O157:H7. The results revealed that for both microorganisms 4% of the examined samples presumed to contain Salmonella and E. coli O157:H7 organisms, according to the colonial character on Harlequin Salmonella ABC agar media and Harlequin SMAC-BCIG agar media. According to biochemical and serological identifications, both organisms could not be detected in the examined samples. A total of 29 (58% samples were positive for antibiotic residues, using the Premi test (a broad-spectrum screening test for the detection of antibiotic residues in meat at or below the maximum residue limits. In addition, aflatoxin B1 was detected in one (2% samples with a concentration of 1.1 μg/kg. The results reflect that there was good hygiene practice for handling and preparation of frozen liver while selling to consumers. However, a high percentage of antibiotic residues reflect ignorance of withdrawal time before slaughtering of animals as well as misuse of antibiotics in veterinary fields. Furthermore, aflatoxin B1 residue was detected in examined frozen liver samples at a concentration below the maximum residual level, which is not enough to cause threat to humans, but it is enough to cause problem if it is eaten regularly reflect contamination of animal feed with aflatoxins.

  19. Frozen soil barriers for hazardous waste confinement

    Energy Technology Data Exchange (ETDEWEB)

    Dash, J.G.; Leger, R. [Univ. of Washington, Seattle, WA (United States); Fu, H.Y. [Univ. of California, Santa Barbara, CA (United States)

    1997-12-31

    Laboratory and full field measurements have demonstrated the effectiveness of artificial ground freezing for the containment of subsurface hazardous and radioactive wastes. Bench tests and a field demonstration have shown that cryogenic barriers are impenetrable to aqueous and non aqueous liquids. As a result of the successful tests the US Department of Energy has designated frozen ground barriers as one of its top ten remediation technologies.

  20. Wildlife Tunnel Enhances Population Viability

    Directory of Open Access Journals (Sweden)

    Rodney van der Ree

    2009-12-01

    Full Text Available Roads and traffic are pervasive components of landscapes throughout the world: they cause wildlife mortality, disrupt animal movements, and increase the risk of extinction. Expensive engineering solutions, such as overpasses and tunnels, are increasingly being adopted to mitigate these effects. Although some species readily use such structures, their success in preventing population extinction remains unknown. Here, we use population viability modeling to assess the effectiveness of tunnels for the endangered Mountain Pygmy-possum (Burramys parvus in Australia. The underpasses reduced, but did not completely remove, the negative effects of a road. The expected minimum population size of a "reconnected" population remained 15% lower than that of a comparable "undivided" population. We propose that the extent to which the risk of extinction decreases should be adopted as a measure of effectiveness of mitigation measures and that the use of population modeling become routine in these evaluations.

  1. Malthus, Boserup and population viability.

    Science.gov (United States)

    Bonneuil, N

    1994-01-01

    The Malthus-Boserup explanatory framework is revisited from the point of view of viability theory. Instead of imposing a univocal relationship between population pressure and level of knowledge, the way technology will change is not determined, it is only constrained. This leads to regard any situation as associated to a set of reachable futures. When no possibility is left for systems to avoid extinction, systems are no longer viable. Hence, the control-phase space can be divided into regions corresponding to gradual danger or security. This point of view allows the introduction of ideas such as incentives to create or to use new knowledge, gives a role to the threatening power of Malthusian checks, and leaves space for a specific variety of behaviors. The Boserupian theme then appears indirectly, emerging from the constraints imposed by the inertia of technological change.

  2. Viability of telework at PROCEMPA.

    Science.gov (United States)

    Fetzner, Maria Amelia de Mesquita

    2003-02-01

    At the end of the 20th century, telework appears as one of the modalities of flexible work, which is related to new organizational structures as well as to increasing use of technology. It revolutionizes the traditional ways of performing work. Its implementation creates a number of questions to be answered by the organizations and the individuals involved. This article presents a case study on the viability of implementing telework at Procempa (The Data Processing Company of the City of Porto Alegre). The case study analyzes the technical, organizational, psychological, legal, and labor union dimensions. As a result of this study, we can identify the organization's stage of readiness for telework, the conditions under which it would be implemented, and the specific issues of an implementation.

  3. The effects of cholesterol on the viability and fertility of bull spermatozoa

    Directory of Open Access Journals (Sweden)

    Polmer Situmorang

    2002-12-01

    Full Text Available This study was conducted to evaluate the effect of cholesterol on the viability and fertility of chilled and deep-frozen bull spermatozoa. Semen was collected by means of artificial vagina, diluted in Tris-Citrat diluent and cooled to 50C for 60 minutes. Following an equiliberation for 4 hours, semen was frozen at 5 cm above surface of liquid nitrogen for 10 minutes. The experiment was 2 x 3 factorial designed with two level of egg yolk (10 and 20% v/v and 3 level of cholesterol (0; 0.5 and 1.0 mg/ml. The viability of spermatozoa was evaluated after the temperature reduced to 50C, stored at 50C for 1, 3 and 7 days and after thawing. For fertility test, cows were artificially inseminated (AI using chilled and frozen semen on the onset or 6 hours of oestrus. Rectal palpation was conducted 3 months after AI to determine the pregnancy. The percentages motile of chilled semen was higher in 0.5 mg/ml than those of 0.0 or 1.0 mg/ml cholesterol but this difference was not significant. After thawing, the effects of cholesterol on the percentage motile was significant (P<0.05. The mean percentage motile was 47.5; 51,5 and 56.0 for 0.0; 0.5 and 1.0 mg/ml cholesterol respectively. The percentage of live sperm and intact apical ridge was higher in cholesterol however this effects was not significant. The effects level of egg yolk and its interaction with cholesterol on the viability was not significant. The percentage of pregnant was higher in 1.0 mg/ml and the mean percentage of pregnant was 45.8; 48.2 and 55.7 for 0.0; 0.5 and 1.0 mg/ml cholesterol respectively. Percentage of pregnant was higher for chilled semen than those of frozen semen (54.3 vs 45.5. In conclusion the addition of 1 mg/ml cholesterol increase the percentage of motile after thawing and pregnancy of cows inseminated with chilled and frozen semen.

  4. Freezing tolerance of sea urchin embryonic cells: Differentiation commitment and cytoskeletal disturbances in culture.

    Science.gov (United States)

    Odintsova, Nelly A; Ageenko, Natalya V; Kipryushina, Yulia O; Maiorova, Mariia A; Boroda, Andrey V

    2015-08-01

    This study focuses on the freezing tolerance of sea urchin embryonic cells. To significantly reduce the loss of physiological activity of these cells that occurs after cryopreservation and to study the effects of ultra-low temperatures on sea urchin embryonic cells, we tested the ability of the cells to differentiate into spiculogenic or pigment directions in culture, including an evaluation of the expression of some genes involved in pigment differentiation. A morphological analysis of cytoskeletal disturbances after freezing in a combination of penetrating (dimethyl sulfoxide and ethylene glycol) and non-penetrating (trehalose and polyvinylpyrrolidone) cryoprotectants revealed that the distribution pattern of filamentous actin and tubulin was similar to that in the control cultures. In contrast, very rare spreading cells and a small number of cells with filamentous actin and tubulin were detected after freezing in the presence of only non-penetrating cryoprotectants. The largest number of pigment cells was found in cultures frozen with trehalose or trehalose and dimethyl sulfoxide. The ability to induce the spicule formation was lost in the cells frozen only with non-penetrating cryoprotectants, while it was maximal in cultures frozen in a cryoprotective mixture containing both non-penetrating and penetrating cryoprotectants (particularly, when ethylene glycol was present). Using different markers for cell state assessment, an effective cryopreservation protocol for sea urchin cells was developed: three-step freezing with a low cooling rate (1-2°C/min) and a combination of non-penetrating and penetrating cryoprotectants made it possible to obtain a high level of cell viability (up to 65-80%). Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Phosphate buffer increases recovery of Escherichia coli O157:H7 from frozen apple juice.

    Science.gov (United States)

    Yamamoto, S A; Harris, L J

    2001-09-01

    It is common practice to dilute food products in 0.1% peptone before microbiological analysis. However, this diluent may not be appropriate for detection of injured organisms present in acidic foods. Shelf-stable unclarified apple juice (pH 3.6) was inoculated with approximately 1 x 10(7) CFU/ml of Escherichia coli O157:H7 and held at 23 +/- 2 degrees C (control) or frozen to -20 +/- 2 degrees C for 24 h to induce injury before sampling. Unfrozen or frozen and thawed juice was diluted 1:1 or 1:10 in 0.1% (wt/vol) peptone (pH 6.1) or 0.1 M phosphate buffer (pH 7.2). Juice samples were plated onto tryptic soy agar with 0.1% (wt/vol) sodium pyruvate (TSAP) to measure survival or onto sorbitol MacConkey agar (SMA) to indicate injury. Counts on TSAP or SMA were the same for control samples held in peptone or phosphate buffer for up to 45 min. However, populations of E. coli in frozen and thawed samples declined rapidly upon dilution in 0.1% peptone. Within 20 min, E. coli underwent a >1-log10 CFU/ml reduction in viability as measured on TSAP and a >2-log10 CFU/ml reduction to below the limit of detection (1.6 or 2.3 log10 CFU/ml) on SMA. In contrast, populations of E. coli in frozen and thawed samples diluted in phosphate buffer did not decrease significantly on TSAP and decreased by diluent results in superior recovery of these organisms on both selective and nonselective plating media.

  6. Effect of magnetic nanoparticle heating on cortical neuron viability.

    Science.gov (United States)

    Rivet, Christopher J; Yuan, Yuan; Gilbert, Ryan J; Borca-Tasciuc, Diana-Andra

    2014-03-01

    Superparamagnetic iron oxide nanoparticles are currently approved for use as an adjunctive treatment to glioblastoma multiforme radiotherapy. Radio frequency stimulation of the nanoparticles generates localised hyperthermia, which sensitises the tumour to the effects of radiotherapy. Clinical trials reported thus far are promising, with an increase in patient survival rate; however, what are left unaddressed are the implications of this technology on the surrounding healthy tissue. Aminosilane-coated iron oxide nanoparticles suspended in culture medium were applied to chick embryonic cortical neuron cultures. Cultures were heated to 37 °C or 45 °C by an induction coil system for 2 h. The latter regime emulates the therapeutic conditions of the adjunctive therapy. Cellular viability and neurite retraction was quantified 24 h after exposure to the hyperthermic events. The hyperthermic load inflicted little damage to the neuron cultures, as determined by calcein-AM, propidium iodide, and alamarBlue® assays. Fluorescence imaging was used to assess the extent of neurite retraction which was found to be negligible. Retention of chick, embryonic cortical neuron viability was confirmed under the thermal conditions produced by radiofrequency stimulation of iron oxide nanoparticles. While these results are not directly applicable to clinical applications of hyperthermia, the thermotolerance of chick embryonic cortical neurons is promising and calls for further studies employing human cultures of neurons and glial cells.

  7. RANCIDITY DEVELOPMENT DURING FROZEN STORAGE OF FILLETS FROM GILTHEAD SEABREAM (Sparus aurata REARED IN ITALY

    Directory of Open Access Journals (Sweden)

    P.P. Gatta

    2013-02-01

    Full Text Available Lipid oxidation indices (Free Fatty Acids-FFA, Peroxide Value-PV and Thiobarbituric Acid-TBA were evaluated in frozen fillets from seabream reared in Italy in: land based facilities (recirculation systems, lagoons or net-cages. Statistically significant differences emerged among seabream sources for all indices. Quality loss related to rancidity seemed to have been affected by both storage time and culturing system.

  8. Morphology and viability of castor bean genotypes pollen grains

    Directory of Open Access Journals (Sweden)

    Maria Selma Alves Silva Diamantino

    2016-01-01

    Full Text Available The objective of this work was to characterize the morphology and viability of the pollen of 15 genotypes of castor bean (Ricinus communis L. and to generate information that can assist in the selection of highly promising male parents for future use in genetic improvement programs aimed at producing seeds for oil extraction. Acetolysis and scanning electron microscopy was used to characterize the morphology of the pollen. The viability of the pollen grains was estimated by in vitro germination and colorimetric analysis (acetocarmine 2% and 2, 3, 5-triphenyltetrazolium chloride 1%. For the in vitro germination, pollen grains were grown in 10 types of solidified culture medium consisting of different concentrations of sucrose, boric acid, calcium nitrate, magnesium sulfate and potassium nitrate. The pollen grains had the following characteristics: medium size, isopolar and subspheroidal shape, radial symmetry, circular ambit, 3-colporate, elongated endoapertures, tectate exine and granulated sexine. The acetocarmine dye overestimated pollen viability. The media M5 and M8 were the most efficient at promoting the germination of pollen grains. The studied genotypes had high levels of viability and can therefore be used as male parents in genetic improvement programs.

  9. A comparison of assays measuring the viability of Legionella ...

    Science.gov (United States)

    Background: The relatively high prevalence of Legionella pneumophila in premise plumbing systems has been widely reported. Published reports indicate Legionella has a comparatively high resistance to chlorine and moreover has the ability to grow in phagocytic amoeba which could provide additional protection in chlorinated drinking water distribution systems. Copper-Silver (Cu-Ag) ionization treatment systems are commercially available for use in large building water systems to help control the risks from Legionella bacteria. The objectives of this study were to develop and optimize Legionella viability assays and use them to investigate the viability of Legionella bacteria after exposure to water treated with coppper and silver ions. Methods: Log phase L. pneumophila cells were used in all experiments and were generated by incubation at 35C for 48 hours in buffered yeast extract broth. Viability assays used included plating on buffered charcoal yeast extract agar to determine the number of culturable cells and treating cells with propidium monoazide (PMA) or ethidium monoazide (EMA) followed by quantitative PCR targeting mip gene of L. pneumophila. The qPCR viability assays were optimized using L. pneumophila inactivated by heat treatment at 65C for 60 min. The effectiveness of Cu-Ag ionization treatment was studied by inoculating L. pneumonia at 105 CFU/mL in water collected directly from a building water system that employed this technology and incubat

  10. Economic viability of anaerobic digestion

    Energy Technology Data Exchange (ETDEWEB)

    Wellinger, A. [INFOENERGIE, Ettenhausen (Switzerland)

    1996-01-01

    The industrial application of anaerobic digestion is a relatively new, yet proven waste treatment technology. Anaerobic digestion reduces and upgrades organic waste, and is a good way to control air pollution as it reduces methane and nitrous gas emissions. For environmental and energy considerations, anaerobic digestion is a nearly perfect waste treatment process. However, its economic viability is still in question. A number of parameters - type of waste (solid or liquid), digester system, facility size, product quality and end use, environmental requirements, cost of alternative treatments (including labor), and interest rates - define the investment and operating costs of an anaerobic digestion facility. Therefore, identical facilities that treat the same amount and type of waste may, depending on location, legislation, and end product characteristics, reveal radically different costs. A good approach for evaluating the economics of anaerobic digestion is to compare it to treatment techniques such as aeration or conventional sewage treatment (for industrial wastewater), or composting and incineration (for solid organic waste). For example, the cost (per ton of waste) of in-vessel composting with biofilters is somewhat higher than that of anaerobic digestion, but the investment costs 1 1/2 to 2 times more than either composting or anaerobic digestion. Two distinct advantages of anaerobic digestion are: (1) it requires less land than either composting or incinerating, which translates into lower costs and milder environmental and community impacts (especially in densely populated areas); and (2) it produces net energy, which can be used to operate the facility or sold to nearby industries.

  11. Direct DNA Extraction from Mycobacterium tuberculosis Frozen Stocks as a Reculture-Independent Approach to Whole-Genome Sequencing

    DEFF Research Database (Denmark)

    Bjorn-Mortensen, K; Zallet, J; Lillebaek, T

    2015-01-01

    Culturing before DNA extraction represents a major time-consuming step in whole-genome sequencing of slow-growing bacteria, such as Mycobacterium tuberculosis. We report a workflow to extract DNA from frozen isolates without reculturing. Prepared libraries and sequence data were comparable...

  12. In vitro development of donated frozen-thawed human embryos in a prototype static microfluidic device: a randomized controlled trial

    NARCIS (Netherlands)

    Kieslinger, Dorit C.; Hao, Zhenxia; Vergouw, Carlijn G.; Kostelijk, Elisabeth H.; Lambalk, Cornelis B.; le Gac, Severine

    Objective: To compare the development of human embryos in microfluidic devices with culture in standard microdrop dishes, both under static conditions. Design: Prospective randomized controlled trial. Setting: In vitro fertilization laboratory. Patient(s): One hundred eighteen donated frozen-thawed

  13. Microbial quality of equine frozen semen.

    Science.gov (United States)

    Corona, A; Cherchi, R

    2009-10-01

    Bacteriological surveillance is little applied in management of equine frozen semen but it is quite important to verify the microbial contamination in order to find out the chance of transmission of pathology to the mare in AI. Authors describe a qualitative and quantitative analysis for bacterial contamination on long time (3-17 years) equine frozen semen stored in liquid nitrogen. The semen checked, produced in Italy and in another Europe country, was cryopreserved in liquid nitrogen inside sealed plastic straws. One hundred and ten straws were checked out for pathogenic and no pathogenic bacteria, aerobes and anaerobes and fungi (moulds and yeasts). The Total Microbial Charge was quite variable with an average of about 1.4 x 10(5)CFU/ml. Mostly the microbial agents identified were fungi (17.5%), Enterobacter-coccus spp. (15%), Pseudomonas spp. (6.25%), Stenothophomonas maltophila (6.25%) and anaerobic bacteria like Propionibacterium granulosum (7.5%) and Clostridium spp. (3.75%). 3.75% were unidentified Gram-negative rod and cocci. Streptococcus spp., Staph. aureus, E. coli, Th. equigenitalis and Mycoplasma spp. were not detected. The most represented species were Enterobacter-coccus spp. (1.1 x 10(5)CFU/ml), St. maltophila (8 x 10(4)CFU/ml) and Pr. granulosum (7 x 10(4)CFU/ml) while yeast and even more moulds were little abundant (4.7 x 10(4) and 3.4 x 10(4)CFU/ml respectively). The knowledge of equine frozen semen microbial quality is essential to check out transmission of venereal disease and improve the quality of cryopreserved germplasm.

  14. Viabilidade, confirmação taxonômica e detecção enzimática de espécies de Acremonium preservadas sob óleo mineral na Coleção de Culturas University Recife Mycology Viability, taxonomic confirmation and enzymatic detection of Acremonium species preserved under mineral oil in the URM Culture Collection

    Directory of Open Access Journals (Sweden)

    Suellen Carvalho de Moura Braz

    2009-02-01

    Full Text Available Enzimas hidrolíticas secretadas por fungos têm um papel importante na patogenicidade das infecções. Objetivando avaliar a atividade enzimática foram testados 31 isolados de Acremonium mantidos na Coleção de Culturas University Recife Mycology. Fragmentos das culturas foram transferidos para caldo glicosado para reativação e posterior crescimento em meio ágar batata dextrose, para verificar viabilidade, pureza e confirmação taxonômica pela observação das características macroscópicas e microscópicas. Para detecção enzimática foram utilizados substratos de caseína do leite e gelatina para protease, amido para amilase e lecitina de soja para fosfolipase. Das 31 culturas, 26 (83,9% mantiveram-se viáveis e 24 (92,3% foram confirmadas taxonomicamente. Das 24 culturas, 12 (50% apresentaram atividade proteásica, duas (16,7% em caseína do leite, uma (8,3% em gelatina e nove (75% em ambos os substratos; 16 (66,7% degradaram amido. Nenhuma cultura apresentou atividade fosfolipásica. Conclui-se que espécies de Acremonium são capazes de produzir enzimas envolvidas na patogenicidade das infecções fúngicas.Hydrolytic enzymes secreted by fungi play an important role in the pathogenesis of infection. With the aim of evaluating the enzymatic activity, 31 isolates of Acremonium stored in the University of Recife Mycology (URM Culture Collection were tested. Culture fragments were transferred to glycoside broth for reactivation and further growth in potato dextrose agar medium in order to investigate viability and purity and to confirm the taxonomy through observing the macroscopic and microscopic characteristics. To detect enzymes, milk casein and gelatin were used as substrates for proteinase, starch for amylase and soy lecithin for phospholipase. Among the 31 cultures, 26 (83.9% remained viable and 24 (92.3% were confirmed taxonomically. Out of these 24 cultures, 12 (50% presented proteinase activity, of which two (16.7% were on

  15. Does the balanced scorecard support organizational viability?

    NARCIS (Netherlands)

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

    2003-01-01

    In this paper we assess whether the balanced scorecard (BSC) supports the necessary functions for organizational viability. To this purpose, we use the viable system model (VSM) as a means to describe the functions required for organizational viability. Then we use the VSM as a template to assess

  16. Pollen viability and membrane lipid composition

    NARCIS (Netherlands)

    Bilsen, van D.G.J.L.

    1993-01-01

    In this thesis membrane lipid composition is studied in relation to pollen viability during storage. Chapter 1 reviews pollen viability, membranes in the dry state and membrane changes associated with cellular aging. This chapter is followed by a study of age-related changes in phospholipid

  17. Different photolysis kinetics at the surface of frozen freshwater vs. frozen salt solutions

    Directory of Open Access Journals (Sweden)

    T. F. Kahan

    2010-11-01

    Full Text Available Reactions at air-ice interfaces can proceed at very different rates than those in aqueous solution, due to the unique disordered region at the ice surface known as the quasi-liquid layer (QLL . The physical and chemical nature of the surfacial region of ice is greatly affected by solutes such as sodium halide salts. In this work, we studied the effects of sodium chloride and sodium bromide on the photolysis kinetics of harmine, an aromatic organic compound, in aqueous solution and at the surface of frozen salt solutions above the eutectic temperature. In common with other aromatic organic compounds we have studied, harmine photolysis is much faster on ice surfaces than in aqueous solution, but the presence of NaCl or NaBr – which does not affect photolysis kinetics in solution – reduces the photolysis rate on ice. The rate decreases monotonically with increasing salt concentration; at the concentrations found in seawater, harmine photolysis at the surface of frozen salt solutions proceeds at the same rate as in aqueous solution. These results suggest that the brine excluded to the surfaces of frozen salt solutions is a true aqueous solution, and so it may be possible to use aqueous-phase kinetics to predict photolysis rates at sea ice surfaces. This is in marked contrast to the result at the surface of frozen freshwater samples, where reaction kinetics are often not well-described by aqueous-phase processes.

  18. 9 CFR 381.158 - Poultry dinners (frozen) and pies.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Poultry dinners (frozen) and pies. 381... Composition § 381.158 Poultry dinners (frozen) and pies. Poultry dinners (frozen) and pies shall meet the... kind indicated Percent Weight (Kind) Pies 14 or 11/8 oz. per 8-oz. pie 1 25 or 2 oz. per 8-oz. pie. 1...

  19. VIABILITY OF THE PROBIOTIC BACTERIA L. ACIDOPHILUS IN DAIRY PRODUCTS

    Directory of Open Access Journals (Sweden)

    Janka Koreňová

    2011-12-01

    Full Text Available A number of health benefits have been claimed for probiotic bacteria such as Lactobacillus acidophilus. Because of the potential health benefits, these organisms are increasingly incorporated into dairy foods. Viability of probiotic bacteria is important in order to provide health benefits. However, many studies have shown low viability of probiotics in market preparations. This study cover selective enumeration and survival of probiotic bacteria L. acidophilus in some dairy drinks. L. acidophilus was found in the range from 106 to 107 CFU.g-1 in five types of fermented milk products containing probiotic cultures. Two investigated products were up to standard according to Regulation of Ministry of Agriculture and Ministry of Health of Slovak Republic.doi: 10.5219/147

  20. Sperm distribution and fertilization after unilateral and bilateral laparoscopic artificial insemination with frozen-thawed goat semen.

    Science.gov (United States)

    Anakkul, Nitira; Suwimonteerabutr, Junpen; Tharasanit, Theerawat; Khunmanee, Sarawanee; Diloksumpan, Paweena; Berg, Debra K; Techakumphu, Mongkol

    2014-11-01

    Generally, laparoscopic artificial insemination (LAI) provides a higher success rate than of cervical insemination in goats. However, the sperm distribution after LAI in goats remains unknown, particularly when frozen-thawed semen is used. This study evaluated the distribution of frozen-thawed goat spermatozoa after LAI and compared the effects of sperm numbers and deposition sites (unilateral and bilateral sites) on pregnancy rate. In experiment 1, the frozen-thawed spermatozoa were stained either with CellTracker Green CMFDA (CT-Green) or CellTracker Red CMPTX (CT-Red), and in vitro evaluations of viability and motility were performed. In experiment 2, the labeled spermatozoa were deposited via LAI into the left (CT-Green) and right (CT-Red) uterine horns (n = 4). After ovariohysterectomy (6 hours after insemination), the distributions of green- and red-colored spermatozoa were assessed via tissue section, flushing, and the oviductal contents were also collected. Experiment 3 was designed to test the pregnancy rates in a group of 120 does after LAI using different numbers of spermatozoa (60 and 120 × 10(6) sperm per LAI) and different deposition sites. The results demonstrated that the fluorochromes used in this study did not impair sperm motility or viability. Frozen-thawed goat spermatozoa can migrate transuterinally after LAI, as evidenced by the observations of both CT-Green- and CT-Red-labeled spermatozoa in both uterine horns. Lower numbers of spermatozoa (60 × 10(6)) that are inseminated unilaterally (either ipsilateral or contralateral to the site of ovulation) can efficiently be used for LAI in goats (with a 56.67% pregnancy rate). Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Cell motility, morphology, viability and proliferation in response to nanotopography on silicon black

    DEFF Research Database (Denmark)

    Lopacinska, Joanna M.; Gradinaru, Cristian; Wierzbicki, Rafal

    2012-01-01

    standard measurements of cell viability, proliferation, and morphology on various surfaces. We also analyzed the motility of cells on the same surfaces, as recorded in time lapse movies of sparsely populated cell cultures. We find that motility and morphology vary strongly with nano-patterns, while...... viability and proliferation show little dependence on substrate type. We conclude that motility analysis can show a wide range of cell responses e. g. over a factor of two in cell speed to different nano-topographies, where standard assays, such as viability or proliferation, in the tested cases show much...

  2. Sensory quality of frozen shrimp Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Inês Maria Barbosa Nunes Queiroga

    2014-09-01

    Full Text Available The objective of this research was to evaluate the sensory quality of the marine shrimp Litopenaeus vannamei grown and stored in a freezer. A sensory analysis consisted of the Quality Index Method (MIQ to review the raw shrimp and Descriptive Analysis (AD in cooked shrimp in samples stored for a period of 90 days, using eight previously trained panelists. Accommodation comprising shrimp filet (100-120 pieces / kg samples were subjected to freezing in liquid nitrogen (- 86 °C, Freezing Tunnel (- 35 °C and Domestic Freezer (- 18 °C. At 0, 30, 60 and 90 days of storage in these freezing systems, determination of pH, water holding capacity (WHC, weight loss during cooking (WLC, shearing force, color, total volatile bases (TVB and thiobarbituric acid reactive substances (TBARS were performed. The attributes manual firmness and softness showed better results in rapid freezing. The color parameters (a *, b * and L *, WHC and WCL were higher at 90 days of storage, no significant losses were observed. Highlighted the strength of greater shear in slowly frozen samples at 90 days, confirming the results reported by the sensory panel. At 90 days of storage, the frozen shrimp showed good sensory quality and physical and chemistry characteristics. The shelf life of this shrimp could be set at about 90 days.

  3. Frozen Microemulsions for MAPLE Immobilization of Lipase

    Directory of Open Access Journals (Sweden)

    Valeria Califano

    2017-12-01

    Full Text Available Candida rugosa lipase (CRL was deposited by matrix assisted pulsed laser evaporation (MAPLE in order to immobilize the enzyme with a preserved native conformation, which ensures its catalytic functionality. For this purpose, the composition of the MAPLE target was optimized by adding the oil phase pentane to a water solution of the amino acid 3-(3,4-dihydroxyphenyl-2-methyl-l-alanine (m-DOPA, giving a target formed by a frozen water-lipase-pentane microemulsion. Fourier transform infrared (FTIR spectroscopy and atomic force microscopy (AFM were used to investigate the structure of MAPLE deposited lipase films. FTIR deconvolution of amide I band indicated a reduction of unfolding and aggregation, i.e., a better preserved lipase secondary structure in the sample deposited from the frozen microemulsion target. AFM images highlighted the absence of big aggregates on the surface of the sample. The functionality of the immobilized enzyme to promote transesterification was determined by thin layer chromatography, resulting in a modified specificity.

  4. Bacteriological profile of raw, frozen chicken nuggets.

    Science.gov (United States)

    Eglezos, Sofroni; Dykes, Gary A; Huang, Bixing; Fegan, Narelle; Stuttard, Ed

    2008-03-01

    The bacteriological profile of raw, frozen chicken nuggets manufactured at a chicken processing facility in Queensland, Australia, was determined. Chicken nuggets are manufactured by grinding poultry, adding premixes to incorporate spices, forming the meat to the desired size and shape, applying a batter and breading, freezing, and packaging. A total of 300 frozen batches were analyzed for aerobic plate count, Escherichia coli, and Salmonella over a period of 4 years. The mean of the aerobic plate count was 5.4 log CFU/g, and counts at the 90th, 95th, and 99th percentiles were 5.7, 5.9, and 6.5 log CFU/g, respectively. The maximum number of bacteria detected was 6.6 log CFU/g. E. coli prevalence was 47%, and of the positive samples, the mean was 1.9 log CFU/g; counts at the 90th, 95th, and 99th percentiles were 2.3, 2.4, and 2.8 log CFU/g, respectively. The maximum number of E. coli was 2.9 log CFU/g. The Salmonella prevalence was 8.7%, and 57.7% of these isolates were typed as Salmonella subspecies II 4,12,[27]:b:[e,n,x] (Sofia), a low-virulence serotype well adapted to Australian poultry flocks. There was a significant relationship (P nuggets.

  5. Analysis of the effect of cryoprotectant medium composition to viability of autologous hematopoietic cells collected by leukapheresis.

    Science.gov (United States)

    Kozlowska-Skrzypczak, M; Kubiak, A; Bembnista, E; Matuszak, P; Komarnicki, M

    2014-10-01

    Cryopreservation of hematopoietic stem cells intended for autologous transplantation is a crucial element of the banking process. Although cryopreservation techniques are well known, improvement is needed. This study was designed to optimize cryopreservation to improve the quantitative and qualitative parameters of hematopoietic stem cells in the material intended for transplantation. We used available opportunities to provide the best quantitative and qualitative parameters of hematopoietic stem cell transplants processed in a closed system. Two hundred forty-eight products of hematopoietic stem cells collected by leukapheresis from patients with lymphoproliferative disorders create the basis of this report. The material was frozen in a controlled-rate freezer and stored in containers in the vapor phase of LN2 (-160°C). The composition of a cryoprotectant medium was modified. For freezing, 192 probes were used with a cryoprotective medium containing 20% dimethyl sulfoxide (DMSO) and enriched RPMI 1640. For 56 samples, we used 20% DMSO in autologous plasma harvested during leukapheresis. Products of hematopoietic stem cells and cryoprotectant medium were combined in a 1:1 ratio. The final number of nuclear cells did not exceed 2 × 10(8)/mL. Analysis was performed after thawing the probes. Viability of nuclear cells has been assessed using the microscopic technique after incubation in Trypan blue and the CD34+ cells by flow cytometry using the 7-aminoactynomycin D. A statistical analysis has been conducted using the Statistica program (StatSoft, Cracow, Poland). The results show that the application of autologous plasma is linked with higher viability of nuclear cells and CD34+ cells. Moreover, statistical analysis of the nuclear cells and CD34+ cells viability differs significantly between groups frozen using RPMI 1640 and autologous plasma (P < .05). To assess the viability of CD34+, cells frozen using RPMI 1640 results showed a large span of at 16.4% to 99

  6. The effect of glucocorticoids on tendon cell viability in human tendon explants

    Science.gov (United States)

    Lui, Wai Ting; Chuen Fu, Sai; Man Lee, Kwong

    2009-01-01

    Background and purpose Previous studies on the culture of human tenocytes have shown that dexamethasone and triamcino-lone reduce cell viability, suppress cell proliferation, and reduce collagen synthesis. However, such cell cultures lack the extracellular matrix and three-dimensional structure of normal tendons, which affects their response to stimuli. We established a human tendon explant culture system and tested the effects of dexamethasone and triamcinolone on cell viability. Methods Primary human tendon explant cultures were prepared from healthy hamstring tendons. Tendon strips were harvested from hamstring tendons and cultured in 24-well plates in Dulbecco’s modification of Eagle’s Medium (DMEM) supplemented with 2% fetal calf serum. The tendon explants were treated with 0 μM (control), 10 μM, or 100 μM dexamethasone sodium phosphate or 0 μM (control), 10 μM, or 100 μM triamcinolone acetonide in DMEM for 96 h. Cell viability was measured by Alamar blue assay before and after glucocorticoid treatment. Results Incubation with 10 μM and 100 μM dexamethasone reduced cell viability in human tendon explants by 35% and 45%, respectively, as compared to a 6% increase in the controls (p = 0.01, mixed-effects ANOVA). Triamcinolone at 10 μM and 100 μM reduced cell viability by 33% and 36%, respectively, as compared to a 9% increase in the controls (p = 0.07, mixed-effects ANOVA). Interpretation Human tendon explant cultures can be used to study the effects of glucocorticoids on human tendon. Dexamethasone and triamcinolone suppress the cell viability of human tendon in its natural 3-dimensional environment with matrix anchorage. Human tendon explant cultures provide a species-specific model for further investigation of the effects of glucocorticoids on the metabolism of the extracellular matrix of human tendon, and on its mechanical properties. PMID:19421908

  7. PROFITABILITY AND VIABILITY OF CATFISH ENTERPRISES IN ...

    African Journals Online (AJOL)

    The inadequacies in the supply of animal protein in the diet of Nigerians have resulted in massive importation of frozen meat, fish and chicken with the consequent loss of scarce foreign exchange. Efforts to boost animal production and to bridge the gap between supply and demand are particularly directed at fish production ...

  8. Terminology for pregnancy loss prior to viability

    DEFF Research Database (Denmark)

    Kolte, A M; Bernardi, L A; Christiansen, O B

    2015-01-01

    Pregnancy loss prior to viability is common and research in the field is extensive. Unfortunately, terminology in the literature is inconsistent. The lack of consensus regarding nomenclature and classification of pregnancy loss prior to viability makes it difficult to compare study results from...... different centres. In our opinion, terminology and definitions should be based on clinical findings, and when possible, transvaginal ultrasound. With this Early Pregnancy Consensus Statement, it is our goal to provide clear and consistent terminology for pregnancy loss prior to viability....

  9. Optimizing recovery of frozen human peripheral blood mononuclear cells for flow cytometry.

    Directory of Open Access Journals (Sweden)

    Bo Langhoff Hønge

    Full Text Available Live peripheral blood mononuclear cells (PBMCs can be frozen and thawed for later analyses by adding and removing a cryoprotectant, such as dimethyl sulfoxide (DMSO. Laboratories across the world use various procedures, but published evidence of optimal thawing procedures is scarce.PBMCs were separated from blood collected from healthy Danish blood donors, and stored at -80°C after adding of DMSO. The essential steps in the thawing procedure were modified and performance was evaluated by flow cytometry with respect to the percentage and total yield of viable PMBCs.The best-performing washing medium was Roswell Park Memorial Institute (RPMI 1640 at 37°C with 20% fetal bovine serum. When using 10 mL washing medium in a 15-mL Falcon tube, samples should be centrifuged for at least 10 minutes at 500 g. We failed to detect any differences between the tested methods of mixing PBMCs with washing medium. Likewise, neither the thawing duration nor centrifugation temperature (20°C and 37°C had any effect. PBMCs could be incubated (rested for up to eight hours in a 37°C 5% CO2 incubator without affecting cell counts, but incubating PBMCs for 16 hours significantly decreased viability and recovery. In general, high viability was not necessarily associated with high recovery.Changing the thawing procedure significantly impacted PBMC viability and live cell recovery. Evaluating both viability and live PBMC recovery are necessary to evaluate method performance. Investigation of differential loss of PBMC subtypes and phenotypic changes during thawing and incubation requires further evaluation.

  10. Optimizing recovery of frozen human peripheral blood mononuclear cells for flow cytometry.

    Science.gov (United States)

    Hønge, Bo Langhoff; Petersen, Mikkel Steen; Olesen, Rikke; Møller, Bjarne Kuno; Erikstrup, Christian

    2017-01-01

    Live peripheral blood mononuclear cells (PBMCs) can be frozen and thawed for later analyses by adding and removing a cryoprotectant, such as dimethyl sulfoxide (DMSO). Laboratories across the world use various procedures, but published evidence of optimal thawing procedures is scarce. PBMCs were separated from blood collected from healthy Danish blood donors, and stored at -80°C after adding of DMSO. The essential steps in the thawing procedure were modified and performance was evaluated by flow cytometry with respect to the percentage and total yield of viable PMBCs. The best-performing washing medium was Roswell Park Memorial Institute (RPMI) 1640 at 37°C with 20% fetal bovine serum. When using 10 mL washing medium in a 15-mL Falcon tube, samples should be centrifuged for at least 10 minutes at 500 g. We failed to detect any differences between the tested methods of mixing PBMCs with washing medium. Likewise, neither the thawing duration nor centrifugation temperature (20°C and 37°C) had any effect. PBMCs could be incubated (rested) for up to eight hours in a 37°C 5% CO2 incubator without affecting cell counts, but incubating PBMCs for 16 hours significantly decreased viability and recovery. In general, high viability was not necessarily associated with high recovery. Changing the thawing procedure significantly impacted PBMC viability and live cell recovery. Evaluating both viability and live PBMC recovery are necessary to evaluate method performance. Investigation of differential loss of PBMC subtypes and phenotypic changes during thawing and incubation requires further evaluation.

  11. Ciliary neurotrophic factor (CNTF) promotes skeletal muscle progenitor cell (MPC) viability via the phosphatidylinositol 3-kinase-Akt pathway.

    Science.gov (United States)

    Hiatt, Kelly; Lewis, Davina; Shew, Mathew; Bijangi-Vishehsaraei, Khadijeh; Halum, Stacey

    2014-12-01

    Muscle progenitor cells (MPCs) are currently being investigated as cellular vectors to deliver neurotrophic factor (NF) for the promotion of re-innervation after axonal injury. Ideally NF delivery in such a model would enhance axonal regeneration while simultaneously promoting MPC viability. To date, insulin-like growth factor 1 (IGF-1) is one of the few NFs known to promote both re-innervation and MPC viability. We herein identify ciliary neurotrophic factor (CNTF) as a factor that promotes MPC viability in culture, and demonstrate CNTF to impart greater viability effects on MPCs than IGF-1. We demonstrate that pharmacological inhibition via LY294002 results in abrogation of CNTF-mediated viability, suggesting that the CNTF-mediated MPC viability benefit occurs via the PI3-Akt pathway. Finally, we employ a genetic model, establishing MPC cultures from mice deficient in class IA PI-3 K (p85α(-/-) ) mice, and demonstrate that the viability benefit imparted by CNTF is completely abrogated in PI-3 K-deficient MPCs compared to wild-type controls. In summary, our investigations define CNTF as a promoter of MPC viability beyond IGF-1, and reveal that the CNTF-mediated MPC viability effects occur via the PI3-Akt pathway. Copyright © 2012 John Wiley & Sons, Ltd.

  12. Marketing functions and determinants of profit among frozen chicken ...

    African Journals Online (AJOL)

    This study attempted to estimate the cost of performing some functions in frozen chicken marketing and determined the major factors affecting the profit level of the marketers. Using data collected from 10 wholesalers and 29 retailers in Ibadan metropolis, the transportation costs per kilogram of frozen chicken were N1.20 ...

  13. Effect of frozen period on the chemical, microbiological and sensory ...

    African Journals Online (AJOL)

    The study was designed to investigate the effect of duration of frozen storage on the chemical, microbiological and sensory profile of Tilapia fish (Sarotherodun galiaenus) collected from a research pond of the Agricultural Development Project, Akure, Nigeria and were subjected to sixty days of frozen storage and analyzed ...

  14. Coagulation Factors Level in Fresh Frozen Plasma in Rwanda ...

    African Journals Online (AJOL)

    Objectives: To determine the level of coagulation factors and inherited inhibitors in Fresh Frozen Plasma (FFP) and to evaluate Prothrombin Time and activated partial thrombin time in fresh frozen plasma. Design: Cross-sectional study. Setting: Jomo Kenyatta University of Agriculture and Technology in Medical Laboratory ...

  15. Calcium and Phosphorus Content Of Exotic, Local and Frozen ...

    African Journals Online (AJOL)

    ... tissue differences in the content of minerals in chicken muscle. The calcium and phosphorus content of the local chicken places it in a very vintage position to compete favorably with frozen and exotic chicken. Key words: Calcium, Phosphorus, local, exotic and frozen chicken. Journal of Agriculture, Forestry and the Social ...

  16. Frozen soil parameterization in a distributed biosphere hydrological model

    Directory of Open Access Journals (Sweden)

    L. Wang

    2010-03-01

    Full Text Available In this study, a frozen soil parameterization has been modified and incorporated into a distributed biosphere hydrological model (WEB-DHM. The WEB-DHM with the frozen scheme was then rigorously evaluated in a small cold area, the Binngou watershed, against the in-situ observations from the WATER (Watershed Allied Telemetry Experimental Research. First, by using the original WEB-DHM without the frozen scheme, the land surface parameters and two van Genuchten parameters were optimized using the observed surface radiation fluxes and the soil moistures at upper layers (5, 10 and 20 cm depths at the DY station in July. Second, by using the WEB-DHM with the frozen scheme, two frozen soil parameters were calibrated using the observed soil temperature at 5 cm depth at the DY station from 21 November 2007 to 20 April 2008; while the other soil hydraulic parameters were optimized by the calibration of the discharges at the basin outlet in July and August that covers the annual largest flood peak in 2008. With these calibrated parameters, the WEB-DHM with the frozen scheme was then used for a yearlong validation from 21 November 2007 to 20 November 2008. Results showed that the WEB-DHM with the frozen scheme has given much better performance than the WEB-DHM without the frozen scheme, in the simulations of soil moisture profile at the cold regions catchment and the discharges at the basin outlet in the yearlong simulation.

  17. Title: Characterizing a Frozen Extrasolar World

    Science.gov (United States)

    Skemer, Andrew J.; Morley, Caroline V.; Allers, Katelyn N.; Geballe, Thomas R.; Marley, Mark S.; Fortney, Jonathan J.; Faherty, Jacqueline K.; Bjoraker, Gordon L.

    2016-01-01

    The recently discovered brown dwarf WISE 0855 presents our first opportunity to study an object outside the Solar System that is nearly as cold as our own gas giant planets. However the traditional methodology for characterizing brown dwarfs-near infrared spectroscopy-is not currently feasible as WISE 0855 is too cold and faint. To characterize this frozen extrasolar world we obtained a 4.5-5.2 micrometers spectrum, the same bandpass long used to study Jupiter's deep thermal emission. Our spectrum reveals the presence of atmospheric water vapor and clouds, with an absorption profile that is strikingly similar to Jupiter. The spectrum is high enough quality to allow the investigation of dynamical and chemical processes that have long been studied in Jupiter's atmosphere, but this time on an extrasolar world.

  18. A method for working frozen rock

    Energy Technology Data Exchange (ETDEWEB)

    Pavlov, A.Yu.; Isayev, V.B.; Klubnichkin, Ye.K.; Rogach, M.S.; Samoylov, Yu.A.

    1983-01-01

    The method includes division into sectors, initial loosening of the rock and subsequent mining of the loosened rock in the operational surfaces by layers in a descending order by parallel passes. To increase operational safety in mining heaps of caked frozen rock, the layer is loosened in one sector, while the loosened rock mass of the same layer is mined in the other sector with its transport to the bank of the tailings heap, where a sector is formed between the cited sectors in which reference marks are installed on the brow of the bank, and subsequent loosening is conducted. A descent is made on one of the sides of the tailings heap from the operational surfaces to the base of the tailings heap.

  19. Stability of frozen marolo pulp during storage

    Directory of Open Access Journals (Sweden)

    Clarissa Damiani

    2013-12-01

    Full Text Available Marolo, also known as araticum or head-to-black, is a globular berry, a species native to the Brazilian savannah. The aim of this study was to evaluate the physical, chemical, and microbiological stability of frozen marolo pulp during 12 months of frozen storage. It was observed that the levels of ash (0.28-0.22%, protein (0.77-0.71%, lipids (1.75-1.73%, carbohydrates (12.1-10.15%, calorie (67.23-59.01 kcal, sucrose (2.50-1.29%, citric acid (435.63-197.5 µg.g-1, tartaric acid (4.38-1.88 µg.g-1 , acetic acid (470.38-279.25 µg.g-1, ascorbic acid (3.00-0.00 µg.g-1, total pectin (0.67-0.39%, pH (3.88-3.83, and b* chromaticity coordinates (24.85-20.53 decreased reduced during storage, whereas the levels of moisture (85.10-87.19%, color parameters (L* 58.89-62.62 and a* 5.37-7.86, reducing sugars (4.53-5.62%, total soluble sugars (7.1-7.36%, soluble solids (7.0-8.4 ºBrix, total acidity (0.9-1.0%, malic acid (514.13-781.25 µg.g-1, soluble pectin (0.16-0.24%, and antioxidant (6.85-37.35% of DPPH discoloration increased over the one-year of storage period. According to the physical, chemical, and microbiological parameters assessed, the product can be stored for 12 months without loss of quality with addition of citric acid as a preservative.

  20. The commercial viability of any intensively cultured fish species ...

    African Journals Online (AJOL)

    spamer

    were analysed using one-way analysis of variance. Tukey's D-test was used to compare means between treatments at p = 0.05 (Snedecor and Cochran 1990). Protein requirements were estimated from total body weight gain, SGR, FCR and PER using the two-slope, broken-line analysis technique (Robbins et al. 1979).

  1. Cell structure and percent viability by a slide centrifuge technique.

    Science.gov (United States)

    Fitzgerald, M G; Hosking, C S

    1982-01-01

    It was found that a slide centrifuge (Cytospin) preparation of a cell suspension allowed a reliable assessment of not only cell structure but also the percentage of non-viable cells. The non-viable cells appeared as "smear" cells and paralleled in number the cells taking up trypan blue. Direct experiment showed the unstained viable cells in a trypan blue cell suspension remained intact in a Cytospin preparation while the cells taking up trypan blue were the "smear" cells. The non-viability of the "smear" cells was confirmed by their inability to survive in culture. Images PMID:7040483

  2. Production of bovine cloned embryos with donor cells frozen at a slow cooling rate in a conventional freezer (-20 degrees C).

    Science.gov (United States)

    Chacón, Liliana; Gómez, Martha C; Jenkins, Jill A; Leibo, Stanley P; Wirtu, Gemechu; Dresser, Betsy L; Pope, C Earle

    2009-11-01

    SummaryUsually, fibroblasts are frozen in dimethyl sulphoxide (DMSO, 10% v/v) at a cooling rate of 1 degrees C/min in a low-temperature (-80 degrees C) freezer (LTF) before storage in liquid nitrogen (LN2); however, a LTF is not always available. The purpose of the present study was to evaluate apoptosis and viability of bovine fibroblasts frozen in a LTF or conventional freezer (CF; -20 degrees C) and their subsequent ability for development to blastocyst stage after fusion with enucleated bovine oocytes. Percentages of live cells frozen in LTF (49.5%) and CF (50.6%) were similar, but significantly less than non-frozen control (88%). In both CF and LTF, percentages of live apoptotic cells exposed to LN2 after freezing were lower (4% and 5%, respectively) as compared with unexposed cells (10% and 18%, respectively). Cells frozen in a CF had fewer cell doublings/24 h (0.45) and required more days (9.1) to reach 100% confluence at the first passage (P) after thawing and plating as compared with cells frozen in a LTF (0.96 and 4.0 days, respectively). Hypoploidy at P12 was higher than at P4 in cells frozen in either a CF (37.5% vs. 19.2%) or in a LTF (30.0% vs. 15.4%). A second-generation cryo-solution reduced the incidence of necrosis (29.4%) at 0 h after thawing as compared with that of a first generation cryo-solution (DMEM + DMSO, 60.2%). The percentage of apoptosis in live cells was affected by cooling rate (CF = 1.9% vs. LFT = 0.7%). Development of bovine cloned embryos to the blastocyst stage was not affected by cooling rate or freezer type.

  3. Different effects of sonoporation on cell morphology and viability

    Directory of Open Access Journals (Sweden)

    Ji-Zhen Zhang

    2012-05-01

    Full Text Available The objective of our study was to investigate changes in cell morphology and viability after sonoporation. Sonoportion was achieved by ultrasound (21 kHz exposure on adherent human prostate cancer DU145 cells in the cell culture dishes with the presence of microbubble contrast agents and calcein (a cell impermeant dye. We investigated changes in cell morphology immediately after sonoporation under scanning electron microscope (SEM and changes in cell viability immediately and 6 h after sonoporation under fluorescence microscope. It was shown that various levels of intracellular calcein uptake and changes in cell morphology can be caused immediately after sonoporation: smooth cell surface, pores in the membrane and irregular cell surface. Immediately after sonoporation, both groups of cells with high levels of calcein uptake and low levels of calcein uptake were viable; 6 h after sonoporation, group of cells with low levels of calcein uptake still remained viable, while group of cells with high levels of calcein uptake died. Sonoporation induces different effects on cell morphology, intracellular calcein uptake and cell viability.

  4. Dormancy, activation and viability of Rhizopus oligosporus sporangiospores.

    Science.gov (United States)

    Thanh, N V; Nout, M J R

    2004-04-15

    Interruption of dormancy to improve viability of Rhizopus oligosporus sporangiospores is crucial for the application of stored starter cultures for fungal (tempe) production. We aimed to assess the extent of dormancy and factors that could result in activation. Whereas heat treatments were unsuccessful, Malt Extract Broth (MEB) showed to be a good activation medium, with 80% of dormant spores being activated as measured by fluorescence microscopy using a fluorescent marker, compared with 11% with the control. Peptone and yeast extract but not glucose played an important role in activating dormant spores. Metabolically active (fluorescent) and swollen spores, followed by germ tubes were obtained after activation in MEB for 25 min., 2 and 4 h, respectively, at 37 degrees C. Simultaneously, some interesting transitions took place. Dormant spores represent 85-90% of the total spores at harvest and after drying. Their number decreased to 21-32% after activation with MEB with a concomitant increase of metabolically active spores. As a result of storage, some dormancy was lost, yielding an increase of active spores from 11.2% at harvest to 28.8% after 3 months storage. Levels of active spores were well correlated with their viability. By activation of dormant spores, their viability increased; levels of viable and active spores were maximum in 1 month old starter (61.7% and 75.9% of total spores, respectively) but gradually decreased with concomitant increase of the number of dead spores.

  5. Viability And Conidial Production Of Entomopathogenic Fungi Penicillium SP.

    Directory of Open Access Journals (Sweden)

    Nurariaty Agus

    2015-01-01

    Full Text Available Abstract Penicillium sp. order Eurotiales class Eurotiomycetes family Trichocomaceae is one of the entomopathogenic fungi that have the potential to be developed as biological control agent of pests.The study aims to determine the viability and spora production of Entomopathogenic fungi Penicillium sp. Experiments was conducted in Pests Identification and Biological Control laboratory Department of Plant Pest and Disease Faculty of Agriculture Hasanuddin University. The fungus Penicillium sp. cultured in a liquid medium and then added chitin as treatment and others without chitin. The spora viability of fungi was observed on 12th and 24th hours while spora production on 3nd 6th 9th and 12th days after application.The results showed that conidial viability of the fungus Penicillium sp. at 24 hours after application was higher if the medium given chitin than without chitin. The conidial production was higher if given chitin than without chitin. It was highest on 12th day reached 143.4 x 106 conidiaml if media given chitin and on 6th day if without chitin 0.50 x 106 conidiaml.

  6. Near viability for fully nonlinear differential inclusions

    National Research Council Canada - National Science Library

    Irina Căpraru; Alina Lazu

    2014-01-01

    .... We establish a viability result under Lipschitz hypothesis on F, that consists in proving the existence of solutions of the differential inclusion above, starting from a given set, which remain...

  7. Poxvirus viability and signatures in historical relics

    National Research Council Canada - National Science Library

    McCollum, Andrea M; Li, Yu; Wilkins, Kimberly; Karem, Kevin L; Davidson, Whitni B; Paddock, Christopher D; Reynolds, Mary G; Damon, Inger K

    2014-01-01

    Although it has been >30 years since the eradication of smallpox, the unearthing of well-preserved tissue material in which the virus may reside has called into question the viability of variola virus decades or centuries...

  8. Intraspecific variation in pollen viability, germination and ...

    African Journals Online (AJOL)

    Oleaceae) cultivars 'Koroneiki', 'Mastoidis' and 'Kalamata' was studied with scanning electron microscopy to identify genotype- distinguishing characters that could be employed for morphological cultivar discrimination. Pollen viability and germination ...

  9. Addition of Vital Wheat Gluten to Enhance the Quality Characteristics of Frozen Dough Products

    Directory of Open Access Journals (Sweden)

    Virginia Giannou

    2016-01-01

    Full Text Available The aim of this study was to enhance the quality and sensory characteristics of bread made from frozen dough. Both white and whole-wheat flour were used. In order to improve dough strength and stability during frozen storage, samples were supplemented with vital wheat gluten at the levels of 2%, 4%, 5%, and 6% of flour weight. The characteristics of baked samples were determined through weight loss, specific volume, crust, and crumb color, texture, and sensory evaluation. Dough behavior at sub-zero temperatures was further examined for control samples and samples with 6% gluten using Differential Scanning Calorimetry (DSC, while their low molecular sugar content (fructose, glucose, sucrose was measured using High Pressure Liquid Chromatography (HPLC, as it can be associated with yeast viability and dough freezing point depression. The most stable samples were those with 4% and 6% gluten (for white flour and those with 4% and 5% gluten (for whole-wheat flour. Gluten addition raised the freezing point of dough samples and preserved low molecular sugar generation after prolonged storage.

  10. Probiotic viability – does it matter?

    OpenAIRE

    Sampo J. Lahtinen

    2012-01-01

    Probiotics are viable by definition, and viability of probiotics is often considered to be a prerequisite for the health benefits. Indeed, the overwhelming majority of clinical studies in the field have been performed with viable probiotics. However, it has also been speculated that some of the mechanisms behind the probiotic health effects may not be dependent on the viability of the cells, and therefore is also possible that also nonviable probiotics could have some health benefits. The eff...

  11. Fungal Spores Viability on the International Space Station

    Science.gov (United States)

    Gomoiu, I.; Chatzitheodoridis, E.; Vadrucci, S.; Walther, I.; Cojoc, R.

    2016-11-01

    In this study we investigated the security of a spaceflight experiment from two points of view: spreading of dried fungal spores placed on the different wafers and their viability during short and long term missions on the International Space Station (ISS). Microscopic characteristics of spores from dried spores samples were investigated, as well as the morphology of the colonies obtained from spores that survived during mission. The selected fungal species were: Aspergillus niger, Cladosporium herbarum, Ulocladium chartarum, and Basipetospora halophila. They have been chosen mainly based on their involvement in the biodeterioration of different substrate in the ISS as well as their presence as possible contaminants of the ISS. From biological point of view, three of the selected species are black fungi, with high melanin content and therefore highly resistant to space radiation. The visual inspection and analysis of the images taken before and after the short and the long term experiments have shown that all biocontainers were returned to Earth without damages. Microscope images of the lids of the culture plates revealed that the spores of all species were actually not detached from the surface of the wafers and did not contaminate the lids. From the adhesion point of view all types of wafers can be used in space experiments, with a special comment on the viability in the particular case of iron wafers when used for spores that belong to B. halophila (halophilic strain). This is encouraging in performing experiments with fungi without risking contamination. The spore viability was lower in the experiment for long time to ISS conditions than that of the short experiment. From the observations, it is suggested that the environment of the enclosed biocontainer, as well as the species'specific behaviour have an important effect, reducing the viability in time. Even the spores were not detached from the surface of the wafers, it was observed that spores used in the

  12. Resurrection of a Bull by Cloning from Organs Frozen without Cryoprotectant in a −80°C Freezer for a Decade

    Science.gov (United States)

    Hoshino, Yoichiro; Hayashi, Noboru; Taniguchi, Shunji; Kobayashi, Naohiko; Sakai, Kenji; Otani, Tsuyoshi; Iritani, Akira; Saeki, Kazuhiro

    2009-01-01

    Frozen animal tissues without cryoprotectant have been thought to be inappropriate for use as a nuclear donor for somatic cell nuclear transfer (SCNT). We report the cloning of a bull using cells retrieved from testicles that had been taken from a dead animal and frozen without cryoprotectant in a −80°C freezer for 10 years. We obtained live cells from defrosted pieces of the spermatic cords of frozen testicles. The cells proliferated actively in culture and were apparently normal. We transferred 16 SCNT embryos from these cells into 16 synchronized recipient animals. We obtained five pregnancies and four cloned calves developed to term. Our results indicate that complete genome sets are maintained in mammalian organs even after long-term frozen-storage without cryoprotectant, and that live clones can be produced from the recovered cells. PMID:19129919

  13. Debating the viability of ethnicity

    Directory of Open Access Journals (Sweden)

    Vilna Bashi

    2004-01-01

    Full Text Available [First paragraph] Immigration and the Political Economy of Home: West Indian Brooklyn and American Indian Minneapolis, 1945-1992. RACHEL BUFF. Berkeley: University of Califomia Press, 2001. xv + 240 pp. (Paper US$ 18.95 Black Cuban, Black American: A Memoir. EVELIO GRILLO. Houston TX: Arte Püblico Press, 2000. xvi + 134 pp. (Paper US$ 13.95 West Indian in the West: Self Representations in an Immigrant Community. PERCY C. HINTZEN. New York: New York University Press, 2001. x + 200pp. (Paper US$ 18.50 Caribbean Families in Britain and the Transatlantic World. HARRY GOULBOURNE & MARY CHAMBERLAIN (eds.. Oxford UK: Macmillan, 2001. xvi + 270 pp. (Paper £15.50 Legacies: The Story of the Immigrant Second Generation. ALEJANDRO PORTES & RUBÉN G. RUMBAUT. Berkeley: University of Califomia Press/ New York: Russell Sage Foundation, 2001. xxiv + 406 pp. (Paper US$ 19.95 "Ethnicity" and its meaning, both as an identity and as a resilient cultural influence, has dominated late twentieth-century social scientific analyses of the process of immigrant incorporation. Perhaps we may mark the crowning of the term with the publication of Glazer and Moynihan's The Melting Pot, one famous tome that "explained" varying "assimilation" outcomes among the "new" (post-1965 newcomers by examining their ethnic culture for flaws or strengths that justified socioeconomic failure or success. Muddying the ensuing policy debate was the use of buzzwords, like mainstream, deviant, assimilated, minority, black matriarch, absent father, and underclass, that were themselves categorizing and hierarchical. The tautology of hierarchically labeling groups and then asking why groups with different labels have different outcomes seems to be perpetually invisible to the parties in the assimilation debate, but the debate itself rages on. Newer scholarship has added a different voice to that debate, arguing that variance in "assimilation" is instead explained by incorporation into

  14. Effects of Fluid Shear Stress on Cancer Stem Cell Viability

    Science.gov (United States)

    Sunday, Brittney; Triantafillu, Ursula; Domier, Ria; Kim, Yonghyun

    2014-11-01

    Cancer stem cells (CSCs), which are believed to be the source of tumor formation, are exposed to fluid shear stress as a result of blood flow within the blood vessels. It was theorized that CSCs would be less susceptible to cell death than non-CSCs after both types of cell were exposed to a fluid shear stress, and that higher levels of fluid shear stress would result in lower levels of cell viability for both cell types. To test this hypothesis, U87 glioblastoma cells were cultured adherently (containing smaller populations of CSCs) and spherically (containing larger populations of CSCs). They were exposed to fluid shear stress in a simulated blood flow through a 125-micrometer diameter polyetheretherketone (PEEK) tubing using a syringe pump. After exposure, cell viability data was collected using a BioRad TC20 Automated Cell Counter. Each cell type was tested at three physiological shear stress values: 5, 20, and 60 dynes per centimeter squared. In general, it was found that the CSC-enriched U87 sphere cells had higher cell viability than the CSC-depleted U87 adherent cancer cells. Interestingly, it was also observed that the cell viability was not negatively affected by the higher fluid shear stress values in the tested range. In future follow-up studies, higher shear stresses will be tested. Furthermore, CSCs from different tumor origins (e.g. breast tumor, prostate tumor) will be tested to determine cell-specific shear sensitivity. National Science Foundation Grant #1358991 supported the first author as an REU student.

  15. Soy milk as a storage medium to preserve human fibroblast cell viability: an in vitro study.

    Science.gov (United States)

    Moura, Camilla Christian Gomes; Soares, Priscilla Barbosa Ferreira; Reis, Manuella Verdinelli de Paula; Fernandes Neto, Alfredo Júlio; Soares, Carlos José

    2012-01-01

    Soy milk (SM) is widely consumed worldwide as a substitute for cow milk. It is a source of vitamins, carbohydrates and sugars, but its capacity to preserve cell viability has not been evaluated. The purpose of the present study was to investigate the efficacy of SM to maintain the viability of human fibroblasts at short periods compared with different cow milks. Human mouth fibroblasts were cultured and stored in the following media at room temperature: 10% Dulbecco's Modified Eagle Medium (DMEM) (positive control group); long shelf-life ultra-high temperature whole cow milk (WM); long shelf-life ultra-high temperature skim cow milk (SKM); powdered cow milk (PM); and soy milk (SM). After 5, 15, 30 and 45 min, cell viability was analyzed using the MTT assay. Data were analyzed statistically by the Kruskal-Wallis test with post-analysis using the Dunn's method (α=0.05). SKM showed the lowest capacity to maintain cell viability in all analyzed times (p<0.05). At 30 and 45 min, the absorbance levels in control group (DMEM) and SM were significantly higher than in SKM (p<0.05). Cell viability decreased along the time (5-45 min). The results indicate that SM can be used as a more adequate storage medium for avulsed teeth. SKM was not as effective in preserving cell viability as the cell culture medium and SM.

  16. Advanced three-dimensional culture of equine intestinal epithelial stem cells.

    Science.gov (United States)

    Stewart, A Stieler; Freund, J M; Gonzalez, L M

    2017-08-09

    Intestinal epithelial stem cells are critical to epithelial repair following gastrointestinal injury. The culture of intestinal stem cells has quickly become a cornerstone of a vast number of new research endeavours that range from determining tissue viability to testing drug efficacy for humans. This study aims to describe the methods of equine stem cell culture and highlights the future benefits of these techniques for the advancement of equine medicine. To describe the isolation and culture of small intestinal stem cells into three-dimensional (3D) enteroids in horses without clinical gastrointestinal abnormalities. Descriptive study. Intestinal samples were collected by sharp dissection immediately after euthanasia. Intestinal crypts containing intestinal stem cells were dissociated from the underlying tissue layers, plated in a 3D matrix and supplemented with growth factors. After several days, resultant 3D enteroids were prepared for immunofluorescent imaging and polymerase chain reaction (PCR) analysis to detect and characterise specific cell types present. Intestinal crypts were cryopreserved immediately following collection and viability assessed. Intestinal crypts were successfully cultured and matured into 3D enteroids containing a lumen and budding structures. Immunofluorescence and PCR were used to confirm the existence of stem cells and all post mitotic, mature cell types, described to exist in the horse intestinal epithelium. Previously frozen crypts were successfully cultured following a freeze-thaw cycle. Tissues were all derived from normal horses. Application of this technique for the study of specific disease was not performed at this time. The successful culture of equine intestinal crypts into 3D "mini-guts" allows for in vitro studies of the equine intestine. Additionally, these results have relevance to future development of novel therapies that harness the regenerative potential of equine intestine in horses with gastrointestinal disease

  17. Effect of inulin and glycerol supplementation on physicochemical properties of probiotic frozen yogurt.

    Science.gov (United States)

    Muzammil, Hafiz Shehzad; Rasco, Barbara; Sablani, Shyam

    2017-01-01

    The present study was designed to investigate the effects of inulin and glycerol supplementation on physicochemical properties of probiotic frozen yogurt. Frozen yogurt was prepared with different types of probiotic ( Lactobacillus acidophilus and Bifidobacterium lactis ) along with yogurt starter culture ( Streptococcus thermophilus and Lactobacillus bulgaricus ). The frozen yogurt mixture was supplemented with inulin (2%, 4%, and 6%) and glycerol (1%, 2%, 3%, and 4%). The results showed that inulin 4% and 6% supplementation increased the overrun by 3% and 5% and the glass transition temperature by 3.3% and 2.8%, and decreased the hardness by 7% and 11%, respectively. Inulin supplementation did not have a significant effect on ice crystal size ( p  > 0.05). Glycerol supplementation increased the stickiness from 2.4% to 18.7%, and decreased the hardness from 8.0% to 14.5% and the glass transition temperature from 2.4% to 34.5%, respectively. Glycerol supplementation did not have a significant effect on overrun or melting rate ( p  > 0.05).

  18. DNA fragmentation in frozen sperm of Equus asinus: Zamorano-Leonés, a breed at risk of extinction.

    Science.gov (United States)

    Cortés-Gutiérrez, E I; Crespo, F; Gosálvez, A; Dávila-Rodríguez, M I; López-Fernández, C; Gósalvez, J

    2008-05-01

    The dynamics of sperm DNA fragmentation (sDF) and sperm viability were analyzed in frozen-thawed sperm samples of Equus asinus (Zamorano-Leonés), a breed at risk of extinction. Sperm DNA fragmentation was assessed using an adaptation of the sperm chromatin dispersion test developed for stallions in five different frozen samples. Sperm were thawed and incubated at different temperatures (37 degrees C, 25 degrees C, and 4 degrees C) and sDF was assessed at different times and compared. The mean sDF after thawing at the beginning of the experiment was 18.20+/-14.77% and did not differ significantly from the results of a neutral comet assay (22.0+/-19.34%). The tendency in the sDF of all donkeys indicated that sperm DNA is more sensitive to breakage when incubated at 37 degrees C than when incubated at 25 degrees C or 4 degrees C. Interestingly, the tendency was not the same when different animals were compared, and differences in sDF dynamics were established among individuals. sDF correlated negatively with sperm viability in some individuals but not in others. From a conservation perspective, sDF analysis may offer a new way to assess sperm quality in endangered breeds in order to identify and select the best semen samples for artificial reproduction purposes. In particular, we recommend for artificial insemination the use of semen samples with a slow increase in sDF with time after thawing.

  19. Viability and molecular authentication of Coccidioides spp. isolates from the Instituto de Medicina Tropical de São Paulo culture collection, Brazil Viabilidade e autenticação molecular de isolados de Coccidioides spp. da coleção de culturas do Instituto de Medicina Tropical de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Sarah Desirée Barbosa Cavalcanti

    2013-02-01

    Full Text Available Coccidioidomycosis is an emerging fungal disease in Brazil; adequate maintenance and authentication of Coccidioides isolates are essential for research into genetic diversity of the environmental organisms, as well as for understanding the human disease. Seventeen Coccidioides isolates maintained under mineral oil since 1975 in the Instituto de Medicina Tropical de São Paulo (IMTSP culture collection, Brazil, were evaluated with respect to their viability, morphological characteristics and genetic features in order to authenticate these fungal cultures. Only five isolates were viable after almost 30 years, showing typical morphological characteristics, and sequencing analysis using Coi-F and Coi-R primers revealed 99% identity with Coccidioides genera. These five isolates were then preserved in liquid nitrogen and sterile water, and remained viable after two years of storage under these conditions, maintaining the same features.Coccidioidomicose é uma doença emergente no Brasil; a manutenção adequada e autenticação de isolados de Coccidioides spp são essenciais para a pesquisa em diversidade genética de micro-organismos, bem como para a compreensão da doença em humanos. Dezessete isolados de Coccidioides preservados em óleo mineral desde 1975 na coleção de culturas do Instituto de Medicina Tropical de São Paulo (IMTSP foram avaliados com relação à viabilidade, características morfológicas e genéticas, com o objetivo de autenticação das culturas fúngicas. Dos 17 isolados, apenas cinco foram viáveis após quase 30 anos mantidos em óleo mineral, apresentando características morfológicas e moleculares típicas do gênero, o sequenciamento utilizando os oligonucleotídeos Coi-F e Coi-R revelou identidade de 99% com isolados de Coccidioides. Estes cinco isolados foram preservados em nitrogênio líquido e água destilada esterilizada, e permaneceram viáveis após dois anos de armazenamento sob estas condições, mantendo

  20. Viability analysis in biological evaluations: Concepts of population viability analysis, biological population, and ecological scale

    Science.gov (United States)

    Gregory D. Hayward; John R. Squires

    1994-01-01

    Environmental protection strategies often rely on environmental impact assessments. As part of the assessment process biologists are routinely asked to evaluate the effects of management actions on plants and animals. This evaluation often requires that biologists make judgments about the viability of affected populations. However, population viability...

  1. Effect of air drying on bacterial viability: A multiparameter viability assessment

    NARCIS (Netherlands)

    Nocker, A.; Fernández, P.S.; Montijn, R.; Schuren, F.

    2012-01-01

    The effect of desiccation on the viability of microorganisms is a question of great interest for a variety of public health questions and industrial applications. Although viability is traditionally assessed by plate counts, cultivation-independent methods are increasingly applied with the aim to

  2. Viability and Resilience of Languages in Competition

    Science.gov (United States)

    Chapel, Laetitia; Castelló, Xavier; Bernard, Claire; Deffuant, Guillaume; Eguíluz, Víctor M.; Martin, Sophie; Miguel, Maxi San

    2010-01-01

    We study the viability and resilience of languages, using a simple dynamical model of two languages in competition. Assuming that public action can modify the prestige of a language in order to avoid language extinction, we analyze two cases: (i) the prestige can only take two values, (ii) it can take any value but its change at each time step is bounded. In both cases, we determine the viability kernel, that is, the set of states for which there exists an action policy maintaining the coexistence of the two languages, and we define such policies. We also study the resilience of the languages and identify configurations from where the system can return to the viability kernel (finite resilience), or where one of the languages is lead to disappear (zero resilience). Within our current framework, the maintenance of a bilingual society is shown to be possible by introducing the prestige of a language as a control variable. PMID:20126655

  3. Probiotic viability – does it matter?

    Directory of Open Access Journals (Sweden)

    Sampo J. Lahtinen

    2012-06-01

    Full Text Available Probiotics are viable by definition, and viability of probiotics is often considered to be a prerequisite for the health benefits. Indeed, the overwhelming majority of clinical studies in the field have been performed with viable probiotics. However, it has also been speculated that some of the mechanisms behind the probiotic health effects may not be dependent on the viability of the cells and, therefore, is also possible that also non-viable probiotics could have some health benefits. The efficacy of non-viable probiotics has been assessed in a limited number of studies, with varying success. While it is clear that viable probiotics are more effective than non-viable probiotics and that, in many cases, viability is indeed a prerequisite for the health benefit, there are also some cases where it appears that non-viable probiotics could also have beneficial effects on human health.

  4. Bupivacaine decreases cell viability and matrix protein synthesis in an intervertebral disc organ model system.

    Science.gov (United States)

    Wang, Dong; Vo, Nam V; Sowa, Gwendolyn A; Hartman, Robert A; Ngo, Kevin; Choe, So Ra; Witt, William T; Dong, Qing; Lee, Joon Y; Niedernhofer, Laura J; Kang, James D

    2011-02-01

    Bupivacaine is a local anesthetic commonly used for back pain management in interventional procedures. Cytotoxic effects of bupivacaine have been reported in articular cartilage and, recently, in intervertebral disc cell culture. However, the relevance of these effects to discs in vivo remains unclear. This study examines the effect of bupivacaine on disc cell metabolism using an organotypic culture model system that mimics the in vivo environment. To assess the effect of bupivacaine on disc cell viability and matrix protein synthesis using an organotypic model system and to determine whether this anesthetic has toxic effects. Mouse intervertebral discs were isolated and maintained ex vivo in an organotypic culture then exposed to clinically relevant concentrations of bupivacaine, and the impact on disc cell viability and matrix proteoglycan (PG) and collagen syntheses were measured in the presence and absence of the drug. Mouse functional spine units (FSUs) were isolated from the lumbar spines of 10-week-old mice. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Total PG and collagen syntheses were determined by measuring the incorporation of radioactive (35)S-sulfate and (3)H-l-proline into PG and collagen, respectively. Organotypic cultures of mouse FSUs were exposed to different concentrations (0%-0.5%) of bupivacaine for variable amounts of time (0-2 hours). Cell viability within disc tissue was quantified by MTT staining and histologic assay. Matrix protein synthesis was measured by incorporation of radioactive (35)S-sulfate (for PG synthesis) and (3)H-l-proline (for collagen synthesis). Untreated mouse disc organs were maintained in culture for up to 1 month with minimal changes in tissue histology, cell viability, and matrix protein synthesis. Exposure to bupivacaine decreased cell viability in a dose- and time-dependent manner. Exposure to bupivacaine at concentrations less than or equal to 0.25% did

  5. Heat and mass transfer in frozen porous media

    NARCIS (Netherlands)

    Loon, van W.

    1991-01-01

    In this thesis processes and parameters associated with heat and mass transfer in frozen porous media both on a theoretical and empirical basis are studied. To obtain the required measurements some existing measuring methods needed to be

  6. Role of frozen section in surgery of benign thyroid

    Directory of Open Access Journals (Sweden)

    Tomaž Štupnik

    2007-04-01

    Full Text Available Background: Fine-needle aspiration biopsy can very reliably discriminate between benign and malignant thyroid tumors. The role of additional intraoperative frozen section to guide intraoperative management was questioned.Methods: A retrospective analysis of all thyroid resections at our department between 2000 and 2004 was performed.Results: 420 patients with benign thyroid lesions underwent surgical resections. 169 lobectomies and 251 total thyroidectomies were performed and 57 (13.5 % thyroid cancers discovered (mostly papillary carcinomas. 54 frozen sections were performed and sensitivity rate of 11.1 % and specificity rate of 97.8 % was estimated.Conclusions: Our data supports the fact that intraoperative frozen section is not sensitive enough to substantially add to preoperative fine-needle aspiration biopsy, even with more selective use. We conclude that routine use of frozen section is not cost-effective in surgery of the benign thyroid and can therefore be abandoned.

  7. A frozen spin polarized target for S134

    CERN Multimedia

    1974-01-01

    The CERN-ETH, Zurich-Helsinki-Imperial College-Southampton Collaboration used a frozen spin polarized target together with the ETH spectrometer magnet to study spin effects (S134). Beam was d31 in South Hall

  8. The biochemical textural and sensory properties of frozen stored ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-08-18

    . Key words: Scallop meats, frozen .... A portion (0.5 ml) of this filtrate was used for the determination of thiobarbituric acid reactive substances ..... mechanism of fish muscle proteins by maltodextrins. Cryobiology, 38: 16-26.

  9. Macro and micro minerals: are frozen fruits a good source?

    Science.gov (United States)

    Spada, Patricia D S; Bortolin, Giovana V; Prá, Daniel; Santos, Carla E I; Dias, Johnny F; Henriques, João A P; Salvador, Mirian

    2010-12-01

    Fruits are rich in minerals, which are essential for a wide variety of metabolic and physiologic processes in the human body. The use of frozen fruits has greatly spread in the last years not only in the preparation of juices, but also as raw material for yogurts, candies, cookies, cakes, ice creams, and children's food. However, up to now there is no data about the mineral profile of frozen fruits. This is the first database to quantify the levels of minerals in 23 samples of frozen fruits, including the most used around the world and some native fruits from the Amazon rainforest in Brazil. Considering the Dietary Reference Intakes, 100g of frozen fruits can provide 0.2 to 2.8% of macro and 2.5 to 100% of microminerals for adults (31-50 years old). Although geographical differences should be considered, these data can help to plan diets and to develop population interventions aiming to prevent chronic diseases.

  10. Explosion Source Characteristics in Frozen and Unfrozen Rock

    National Research Council Canada - National Science Library

    Bonner, Jessie L; Leidig, Mark R; Murphy, Katherine; Dougherty, Sara L; Martin, Randolph J

    2008-01-01

    .... Laboratory studies have demonstrated that frozen rock is significantly stronger than unfrozen rock, and it has been hypothesized that this increased strength, due to ice in the pores and cracks, can alter seismic yield...

  11. Frozen Dynamics and Insulation of Water at the Lipid Interface

    NARCIS (Netherlands)

    Bakulin, A.A.; Cringus, D.; Pshenichnikov, M.S.; Wiersma, D.A.; Corkum, P; DeSilvestri, S; Nelson, KA; Riedle, E

    2009-01-01

    2D IR correlation spectroscopy reveals extremely slow dynamics and splitting of the OH-stretching mode of water in anionic micelles. Water at the lipid interface behaves as if the molecules were isolated in a "frozen" environment.

  12. Use of fluorescent redox indicators to evaluate cell proliferation and viability

    DEFF Research Database (Denmark)

    Rasmussen, E.S.

    1999-01-01

    The performance of two cell viability test kits based on the use of redox indicators yielding fluorescent products, the AlamarBlue assay and a resazurin-based in vitro toxicology assay kit from Sigma, was compared in the present study. Cultures of human neonatal foreskin fibroblasts were exposed...... to equal concentrations of the two dye solutions in the cell culture media. The fluorescence intensities of the cell culture media obtained in response to cell proliferation with the two dyes showed a pronounced similarity. Both dyes were noncytotoxic to cell cultures with high initial cell densities...... components were tentatively identified as resazurin and resorufin. The AlamarBlue assay has gained wide application as a cell viability indicator that allows continuous monitoring of cell proliferation or cytotoxicity in human and animal cells, bacteria, and fungi, but no studies with the deliberate use...

  13. Production of a Functional Frozen Yogurt Fortified with Bifidobacterium spp.

    OpenAIRE

    Amro Abdelazez; Zafarullah Muhammad; Qiu-Xue Zhang; Zong-Tao Zhu; Heba Abdelmotaal; Rokayya Sami; Xiang-Chen Meng

    2017-01-01

    Frozen dairy products have characteristics of both yogurt and ice cream and could be the persuasive carriers of probiotics. Functions of the frozen yogurt containing viable bifidobacterial cells are recognized and favored by the people of all ages. We developed a kind of yogurt supplemented by Bifidobacterium species. Firstly, five strains of Bifidobacterium spp. (Bifidobacterium bifidum ATCC 11547, Bifidobacterium longum ATCC 11549, Bifidobacterium infantis ATCC 11551, Bifidobacterium adoles...

  14. Should Patients With Frozen Shoulder Be Screened for Diabetes Mellitus?

    Science.gov (United States)

    Safran, Ori; El-Haj, Madi; Leibowitz, Gil; Beyth, Shaul; Furman, Zohar; Milgrom, Charles; Kandel, Leonid

    2017-07-01

    Idiopathic frozen shoulder (nontraumatic) is commonly encountered in patients between the ages of 35 and 60 years in general orthopaedic practice. While the prevalence of frozen shoulder among the general population is estimated to be between 2% and 4%, a significantly higher prevalence of 10% to 22% has been reported in patients with diabetes mellitus. Since diabetic patients are more prone to develop frozen shoulder than nondiabetics, the question arises as to whether patients diagnosed as having idiopathic frozen shoulder are at greater risk to develop diabetes mellitus and should be routinely screened for this condition. To compare the prevalence of diabetes mellitus and prediabetes among patients diagnosed with idiopathic frozen shoulder who are not known to have either diabetes mellitus or prediabetic conditions with that of an age-matched group from the general population. Case series; Level of evidence, 4. Patients at a shoulder clinic with a diagnosis of idiopathic frozen shoulder were asked to participate in the study if they were aged between 35 to 60 years and had no known previous diagnosis of diabetes mellitus or prediabetic conditions. These patients underwent a 2-hour oral glucose tolerance test. According to their fasting and 2-hour plasma glucose levels, patients were diagnosed as normal glucose tolerance, prediabetic, or diabetic. Findings were matched with the prevalence in an age-matched general population. Fifty patients completed the test. Four patients with idiopathic frozen shoulder (8%) were found to be prediabetic. No patient was found to be diabetic. All 4 patients reported a history of diabetes in their parents or siblings. Patients diagnosed with idiopathic frozen shoulder who are 60 years or younger and are not known diabetics have a similar probability of having diabetes or prediabetes to an age-matched population. No routine diabetic workup is warranted specifically for these patients.

  15. Biochemical effects on long-term frozen human costal cartilage

    Energy Technology Data Exchange (ETDEWEB)

    Santin, Stefany P.; Martinho Junior, Antonio C.; Yoshito, Daniele; Soares, Fernando A.N.; Mathor, Monica B., E-mail: mathor@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2011-07-01

    Currently, the progresses on treatment of musculoskeletal diseases with the evolving of artificial implants and the success of tissue transplantation between genetically different individuals have conducted to an increase in radiosterilization. Regarding to tissue transplantation, it is essential to have sterile tissue and many tissue banks use radiosterilization as an effective method to sterilize these tissues. However, high doses of ionizing radiation and the preservation method may induce structural modifications in the tissues, as degradation of structural scaffold, decreasing its mechanical properties. Particularly, cartilage have been preserved in high concentrations of glycerol or deep-frozen at -70 degree C for storage after radiosterilization. Therefore, it is important to study the modifications induced in cartilage by preservation methods and by radiosterilization to determine the appropriated parameters for high quality of human allografts. Costal cartilages were obtained from cadaveric donors and were frozen at -20 degree C for 2 years long in order to compare with previous studies for fresh, deep-frozen and glycerolised cartilages. The mechanical tests were carried out in a universal testing machine until sample failure. According our results, there is no significant statistical difference between stress at break of fresh, long-term - 20 degree C frozen cartilages and deep-frozen cartilage. This early result suggests, regarding to tensile property, that long-term - 20 degree C frozen cartilages corresponds to glycerolised costal cartilages irradiated with 25 kGy or deep-frozen cartilages irradiated with 25 and 50 kGy. Thus, this long-term frozen cartilages may be used for tissue banks, but more studies about effects of ionizing radiation are necessary. (author)

  16. Effect of the neurosphere size on the viability and metabolism of ...

    African Journals Online (AJOL)

    The objective of this study was to investigate the metabolic activity and viability of the mouse neural stem/progenitor cells (NSPCs) affected by the size of neurospheres. NSPCs dissociated from the forebrain of embryonic 14 days (E14) mice were cultured in flask for 120 h. During cultivation, the diameter distribution of ...

  17. Pedagogical Technology of Improving the Students' Viability Levels in the Process of Mastering Foreign Language

    Science.gov (United States)

    Dmitrienko, Nadezhda; Ershova, Svetlana; Konovalenko, Tatiana; Kutsova, Elvira; Yurina, Elena

    2015-01-01

    The article points out that the process of mastering foreign language stimulates students' personal, professional and cultural growth, improving linguistic, communicative competences and viability levels. A proposed pedagogical technology of modeling different communicative situations has a serious synergetic potential for students' self organized…

  18. Effects of supplementation of iodixanol to semen extender on quality and fertilization ability of frozen-thawed Thai native bull sperm.

    Science.gov (United States)

    Chuawongboon, Phirawit; Sirisathien, Saksiri; Pongpeng, Jatuporn; Sakhong, Denpong; Nagai, Takashi; Vongpralub, Thevin

    2017-09-01

    This study investigates the effects of iodixanol supplementation in varied concentrations to Tris egg yolk (TEY) extender on the quality and fertilization ability of frozen-thawed sperm of Thai native bulls. Each ejaculate was divided into four different groups, as follows: sperm were treated with TEY extender (control group) and TEY extender supplemented with three different concentrations of iodixanol (1.25%, 2.50% and 5.00%). Semen straws were frozen in liquid nitrogen vapor. After thawing, sperm motility characteristics, viability, plasma membrane integrity and acrosome integrity were determined. Also, frozen-thawed spermatozoa from all groups were used for in vitro fertilization and artificial insemination (AI) in natural estrus Thai native cows. The results showed that the post-thaw quality of the 2.50% iodixanol group was superior to the other iodixanol groups (P fertilization ability and pregnancy rate after AI (P > 0.05). It can be concluded that the supplementation of 2.50% iodixanol extender significantly improves the progressive motility, viability, plasma membrane integrity and acrosome integrity of cryopreserved semen from Thai native bulls, but it has no beneficial effect on in vitro fertilization ability and pregnancy rate after AI. © 2017 Japanese Society of Animal Science.

  19. IN VITRO ASSESSMENT OF EXPOSURE TO NONYLPHENOL ON VIABILITY OF BOVINE SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    Jana Lukáčová

    2014-02-01

    Full Text Available Nonylphenol (NP is a toxic xenobiotic compound classified as an endocrine disruptor that bioaccumulates in the body and causes endocrine disruption. NP can result in male reproductive dysfunction, altered testicular development and decreased male fertility. The target of this in vitro study was to determine the effect of NP as an endocrine disruptor on the viability of spermatozoa. We examined the dose- and time-dependent effect of nonylphenol (1, 10, 100 and 200 µg/mL dissolved either in 0.1% dimethyl sulfoxide (DMSO and 0.1% ethanol on the viability of bovine spermatozoa after 6 h of in vitro cultivation. The viability of bovine spermatozoa was detected by the MTT cytotoxicity assay. The viability in groups with NP dissolved in 0.1% DMSO was significantly (P 10 µg/mL of NP and was decreased significantly (P<0.001 in all experimental groups with NP dissolved in 0.1% ethanol. After 6 h of culture the MTT assay proved a negative effect of all NP doses on the cell viability. The lowest survival of spermatozoa was determined after the addition of 200 µg/mL of NP. The obtained data indicate that the negative effect of NP on the viability must be seriously considered in the case of exposure to NP in animals and humans.

  20. Economic Viability and Marketing Strategies of Periwinkle ...

    African Journals Online (AJOL)

    The economic viability and marketing strategies of periwinkle species in twelve major markets across Rivers State Nigeria were investigated using structured questionnaires. The results indicated that marketing strategies are enroute, through harvesters (collectors), to wholesalers (those who purchase in small quantities ...

  1. Extending the viability of sea urchin gametes.

    Science.gov (United States)

    Spiegler, M A; Oppenheimer, S B

    1995-04-01

    The sea urchin is the material of choice for studying many early developmental events. Methods to extend the viability of sea urchin gametes have not received much attention, but it is well known that the eggs are easily damaged by freezing. This study was designed to extend the viability of Lytechinus pictus eggs and sperm without freezing. Gamete viability measurements were based on relative numbers of fertilized vs unfertilized eggs, percentage fertilization, and on observations of embryonic development. Results indicate that gametes can be stored longer and at lower temperatures than previously described. Sperm were consistently kept viable for at least 12 days with little decrease in viability when stored in glass test tubes or plastic petri dishes and submerged in ice inside a refrigerator at 0 +/- 1 degree C. In one experiment, sperm stored in glass test tubes on ice remained viable up to 20 days after extraction. Eggs were maintained from 1 to 7 days, rather than the 1 day or so previously reported, when stored in glass test tubes submerged in ice in a refrigerator at 0 +/- 1 degree C. Results of egg and sperm experiments varied at different times in the season. Such variations may be caused by seasonal cytoplasmic changes, population differences, or the time mature individuals were maintained unfed in aquaria prior to use. Results from this study should be useful for a variety of research, mariculture, and teaching applications in which sea urchin supplies are limited or when the same gamete population is required for subsequent experiments.

  2. Incorporating evolutionary processes into population viability models

    NARCIS (Netherlands)

    Pierson, J.C.; Beissinger, S.R.; Bragg, J.G.; Coates, D.J.; Oostermeijer, J.G.B.; Sunnucks, P.; Schumaker, N.H.; Trotter, M.V.; Young, A.G.

    2015-01-01

    We examined how ecological and evolutionary (eco-evo) processes in population dynamics could be better integrated into population viability analysis (PVA). Complementary advances in computation and population genomics can be combined into an eco-evo PVA to offer powerful new approaches to understand

  3. Assessment of myocardial viability using PET

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Seok Nam [College of Medicine, Ajou University, Suwon (Korea, Republic of)

    2005-02-15

    The potential for recovery of left ventricular dysfunction after myocardial revascularization represents a practical clinical definition for myocardial viability. The evaluation of viable myocardium in patients with severe global left ventricular dysfunction due to coronary artery disease and with regional dysfunction after acute myocardial infarction is an important issue whether left ventricular dysfunction may be reversible or irreversible after therapy. If the dysfunction is due to stunning or hibernation, functional improvement is observed. But stunned myocardium may recover of dysfunction with no revascularization. Hibernation is chronic process due to chronic reduction in the resting myocardial blood flow. There are two types of myocardial hibernation; 'functional hibernation' with preserved contractile reserve and 'structural hibernation' without contractile reserve in segments with preserved glucose metabolism. This review focus on the application of F-18 FDG and other radionuclides to evaluate myocardial viability. In addition the factors influencing predictive value of FDG imaging for evaluating viability and the different criteria for viability are also reviewed.

  4. Viability of mesenchymal stem cells during electrospinning

    Directory of Open Access Journals (Sweden)

    G. Zanatta

    2012-02-01

    Full Text Available Tissue engineering is a technique by which a live tissue can be re-constructed and one of its main goals is to associate cells with biomaterials. Electrospinning is a technique that facilitates the production of nanofibers and is commonly used to develop fibrous scaffolds to be used in tissue engineering. In the present study, a different approach for cell incorporation into fibrous scaffolds was tested. Mesenchymal stem cells were extracted from the wall of the umbilical cord and mononuclear cells from umbilical cord blood. Cells were re-suspended in a 10% polyvinyl alcohol solution and subjected to electrospinning for 30 min under a voltage of 21 kV. Cell viability was assessed before and after the procedure by exclusion of dead cells using trypan blue staining. Fiber diameter was observed by scanning electron microscopy and the presence of cells within the scaffolds was analyzed by confocal laser scanning microscopy. After electrospinning, the viability of mesenchymal stem cells was reduced from 88 to 19.6% and the viability of mononuclear cells from 99 to 8.38%. The loss of viability was possibly due to the high viscosity of the polymer solution, which reduced the access to nutrients associated with electric and mechanical stress during electrospinning. These results suggest that the incorporation of cells during fiber formation by electrospinning is a viable process that needs more investigation in order to find ways to protect cells from damage.

  5. Viability of smallholder dairying in Wedza, Zimbabwe.

    Science.gov (United States)

    Zvinorova, Plaxedis Ivy; Halimani, Tinyiko Edward; Mano, Renneth T; Ngongoni, Nobbert Takarwirwa

    2013-04-01

    Viability differences in smallholder dairy farming are a result of differences in access to markets and services. It is hypothesized that innovations that improve productivity and market linkages also improve returns and viability. The viability of smallholder dairying in Wedza was characterised by interviewing 52 households using semi-structured questionnaires. Information on demographics, production, marketing, livestock numbers, assets and constraints was obtained. Farmers were resource-constrained with differences in access to resources. The highly resourced farmers had higher milk output and numbers of livestock. Almost 40 % of the households were female-headed, and these dominated the poor category. Household sizes ranged from 4 to 13 persons. Milk off-take was low (3.7 ± 0.53 l/cow/day), due to various constraints. Only rich farmers had viable enterprises in purely financial terms. Per litre cost of milk was more than selling price (US$0.96) for most farmers except the relatively rich. Operating ratios were 1.7, 0.6, 1.4 and 1.1 for the poor, rich, sub-centre and milk collection centre farmers, respectively. This means incomes from the dairy activities did not cover costs. Sensitivity analysis indicated that increases in total variable costs and labour reduced returns. Milk production and viability were influenced by access to resources and markets.

  6. Pollen viability in Quercus robur L.

    Directory of Open Access Journals (Sweden)

    Batos Branislava

    2017-01-01

    Full Text Available The variability of viability (germination rate and the length of pollen tubes of fresh pedunculate oak (Quercus robur L. pollen grains was studied in vitro on a medium containing 15% sucrose. Spatial variability was studied by sampling fresh pollen grains from a total of thirteen trees at four different sites in the area of Belgrade (Košutnjak, Banovo Brdo, Ada Ciganlija and Bojčin Forest in a single year (2011. In order to assess temporal variability and determine the effects of climate change on a small time scale, we studied the viability of the pollen grains collected from one tree at the Banovo Brdo site in six different years (2004, 2005, 2006, 2007, 2011 and 2012. Interindividual variability was tested on the pollen grains sampled from eight trees at Ada Ciganlija in 2004. The percentage values of the pollen grain germination rate and the pollen tube length showed no statistically significant differences between the sites. However, the studied characteristics of the pollen grain viability (germination rate and pollen tube length showed statistically significant differences in both temporal (between the pollen collection years and interindividual variability. This type of research makes a valuable contribution to pedunculate oak breeding programs through the identification of trees with stable production and a good quality of pollen. Furthermore, it can be important in defining the patterns of spatial, temporal and individual variability of pollen grain viability under the influence of climate factors, which are showing compelling changing trends from year to year.

  7. Measuring flow velocity on frozen and non-frozen slopes of black soil through leading edge method

    Directory of Open Access Journals (Sweden)

    Chao Chen

    2017-09-01

    Full Text Available Flow velocity is a major parameter related to hillslope hydrodynamics erosion. This study aims to measure flow velocity over frozen and non-frozen slopes through leading edge method before being calibrated with accurate flow velocity to determine the correct coefficient for convenience of flow velocity measurement. Laboratory experiments were conducted on frozen and non-frozen soil slopes with flumes involving four slope gradients of 5°, 10°, 15°, and 20°and three flow rates of 1, 2, and 4 L/min with a flume of 6 m long and 0.1 m wide. The measurements were made with a stopwatch to record the time duration that the water flow ran over the rill segments of 2, 4 and 6 m long. Accurate flow velocity was measured with electrolyte trace method, under pulse boundary condition. The leading edge and accurate flow velocities were used to determine the correction coefficient to convert the former to the latter. Results showed that the correction coefficient on frozen soil slope was 0.81 with a coefficient of determination (R2 of 0.99. The correction coefficient on non-frozen soil slope was 0.79 with R2 of 0.98. A coefficient of 0.8 was applicable to both soil surface conditions. The accurate velocities on the four frozen black soil slopes were approximately 30%, 54%, 71%, and 91% higher than those on non-frozen soil slopes. By contrast, the leading edge flow velocities on the frozen soil slopes were 23%, 54%, 67%, and 84% higher than those on non-frozen soil slopes. The flow velocities on frozen soil slopes increased with flow rate at all four slopes, but they increased from 5 to 15° before getting stabilized. Therefore, rill flow velocity can be effectively measured with leading edge method by multiplying the leading edge velocity with a correction coefficient of 0.80. This study provides a strategy to measure rill flow velocity for studies on soil erosion mechanisms.

  8. In vitro pollen germination and pollen viability in passion fruit (Passiflora spp.

    Directory of Open Access Journals (Sweden)

    Taliane Leila Soares

    2013-12-01

    Full Text Available The use of Passiflora species for ornamental purposes has been recently developed, but little is known about pollen viability and the potential for crossing different species. The objective of this study was to evaluate the pollen viability of six Passiflora species collected from different physiological stages of development through in vitro germination and histochemical analysis using dyes. The pollen was collected in three stages (pre-anthesis, anthesis and post-anthesis. Three compositions of culture medium were used to evaluate the in vitro germination, and two dyes (2,3,5-triphenyltetrazolium chloride, or TTC, and Lugol's solution were used for the histochemical analysis. The culture medium containing 0.03% Ca(NO3 4H2O, 0.02% of Mg(SO4 .7H2O, 0.01% of KNO3, 0,01% of H3BO3, 15% sucrose, and 0.8% agar, pH 7.0, showed a higher percentage of pollen grains germinated. Anthesis is the best time to collect pollen because it promotes high viability and germination. The Lugol's solution and TTC dye overestimated the viability of pollen, as all accessions showed high viability indices when compared with the results obtained in vitro.

  9. Psychrophilic and Mesophilic Fungi in Frozen Food Products

    Science.gov (United States)

    Kuehn, Harold H.; Gunderson, Millard F.

    1963-01-01

    The mold flora of certain frozen pastries and chicken pies was investigated. Molds were determined qualitatively or quantitatively, or both, by preparing pour plates of the blended product and incubating the plates at various temperatures. The mesophilic fungal flora developed on plates incubated at 10 and 20 C, whereas psychrophilic fungi were obtained on plates incubated at 0 and 5 C. About 2,000 cultures of fungi, representing about 100 different species, were isolated from various products. Four different brands of blueberry, two brands of cherry pastries, two brands of apple, and one brand of raspberry pastries were examined. In addition, two brands of chicken pies were studied. Blueberry pastries had a much higher total fungal population than the other products, although different brands of blueberry pastries varied considerably. Blueberry pastries had from 347 to 1,586 psychrophilic fungi per g. Cherry pastries had about 70 to 110 psychrophiles per g, and apple pastries had 19 to 92 psychrophiles per g. Chicken pies contained very few psychrophilic fungi, about 15 per g. Aureobasidium pullulans was recovered most frequently. About 90% of the psychrophilic fungi found in blueberry products was A. pullulans. Depending upon the brand of cherry pastry, either Phoma spp. or A. pullulans was the most common fungus present. Apple pastries also displayed brand variation, but were unique in having many mesophilic aspergilli. This genus was generally absent from other products. The Penicillium content of apple pastries was also rather high; 50% of the psychrophilic flora was represented by this genus. The psychrophilic fungal flora of chicken pies was composed primarily of penicillia (50%) and Chrysosporium pannorum (46%). PMID:13927344

  10. Non-disruptive measurement system of cell viability in bioreactors

    Science.gov (United States)

    Rudek, F.; Nelsen, B. L.; Baselt, T.; Berger, T.; Wiele, M.; Prade, I.; Hartmann, P.

    2016-04-01

    Nutrient and oxygen transport, as well as the removal of metabolic waste are essential processes to support and maintain viable tissue. Current bioreactor technology used to grow tissue cultures in vitro has a fundamental limit to the thickness of tissues. Based on the low diffusion limit of oxygen a maximum tissue thickness of 200 μm is possible. The efficiency of those systems is currently under investigation. During the cultivation process of the artificial tissue in bioreactors, which lasts 28 days or longer, there are no possibilities to investigate the viability of cells. This work is designed to determine the influence of a non-disruptive cell viability measuring system on cellular activity. The measuring system uses a natural cellular marker produced during normal metabolic activity. Nicotinamide adenine dinucleotide (NADH) is a coenzyme naturally consumed and produced during cellular metabolic processes and has thoroughly been studied to determine the metabolic state of a cell. Measuring the fluorescence of NADH within the cell represents a non-disruptive marker for cell viability. Since the measurement process is optical in nature, NADH fluorescence also provides a pathway for sampling at different measurement depths within a given tissue sample. The measurement system we are using utilizes a special UV light source, to excite the NADH fluorescence state. However, the high energy potentially alters or harms the cells. To investigate the influence of the excitation signal, the cells were irradiated with a laser operating at a wavelength of 355 nm and examined for cytotoxic effects. The aim of this study was to develop a non-cytotoxic system that is applicable for large-scale operations during drug-tissue interaction testing.

  11. Lactose digestion from flavored and frozen yogurts, ice milk, and ice cream by lactase-deficient persons.

    Science.gov (United States)

    Martini, M C; Smith, D E; Savaiano, D A

    1987-10-01

    Lactose digestion from and tolerance to flavored and frozen yogurts, ice cream, and ice milk were evaluated (20 g lactose/meal) in lactase-deficient subjects by use of breath hydrogen techniques. Unflavored yogurt caused significantly less hydrogen production than milk (37 vs 185 delta ppm X h, n = 9). Flavored yogurt was intermediate (77 delta ppm X h). Subjects were free of symptoms after consuming flavored and unflavored yogurts. Of seven commercial yogurts tested, all contained significant levels of microbial beta-galactosidase (beta-gal). In addition, eight subjects were fed meals of milk, ice milk, ice cream, and frozen yogurts with and without cultures containing high levels of beta-gal. Peak hydrogen excretion after consumption of frozen yogurt with high beta-gal was less than one-half of that observed after the other five test meals and intolerance symptoms were absent. Tolerance to frozen yogurt, produced under usual commercial procedures, was found to be similar to that of ice milk and ice cream.

  12. [Repair mechanism of frozen sublethally damaged Staphylococcus aureus].

    Science.gov (United States)

    Huang, Zhongmin; Lv, Haipeng; Ai, Zhilu; Wang, Na; Xie, Xinhua; Fan, Huiping; Pan, Zhili; Suo, Biao

    2015-11-04

    To study the repair mechanisms of frozen sublethally damaged Staphylococcus aurous cells. We resuscitated frozen sublethally damaged S. aureus at 37 degrees C for different time within 3 h. Meanwhile, we compared the morphological changes of the frozen sublethally damaged cells after 1 h of resuscitation using transmission electron microscopy assay (TEM). The expressions of the transcriptional attenuator MsrR (msrR), iron (Fe3+) ABC transporter ATP-binding protein (fhuC), and cytochrome b (cytB) genes were quantitatively analyzed by real-time fluorescence quantitative PCR (Real-time PCR) method. The content of cells outside leakage, active oxygen (ROS), and superoxide dismutase (SOD) activity were also determined by ultraviolet spectrophotometry. More than 99% of the frozen sublethally damaged S. aureus repaired after 3 h. The resuscitated cells expressed an equal resistance to high concentration of NaCl. Real-time PCR results showed that the msrR and fhuC genes expressions were down-regulated, whereas the cytB gene expression was up-regulated significantly. The frozen sublethally damaged S. aureus cellar surface ultrastructure significant changed during resuscitation. The cell surface became compact and sturdy from smooth and transparent. The cell leakage rate of ultraviolet absorption material gradually decreased. Meanwhile, the intracellular ROS level declined along with the decrease of SOD activity. Frozen sublethally damaged cells may regain the capability of resistance to high salt stress by repairing cell membrane integrity, reducing the content of ROS through gene regulation, inhibiting the toxicity of active oxygen to the cells. Meanwhile, the regulation of metabolism related genes (cytB) provides the energy for the requirement of cells, therefore, the frozen sublethally damaged cells were repaired finally.

  13. Morphology and Viability of Pleistocene Microbiota from the CRREL Permafrost Tunnel Near Fox, Alaska

    Science.gov (United States)

    Hoover, Richard B.

    2000-01-01

    The U. S. Army Cold Regions Research and Engineering Laboratory maintains the CRREL Permafrost Tunnel at Fox, Alaska (-10 miles north of Fairbanks.) The active microbial ecosystems and the cryopreserved anabiotic viable microorganisms and dead microbial remains and biomarkers frozen within the permafrost and ice of the CRREL Permafrost Tunnel are of direct relevance to Astrobiology. Microbial extremophiles from permafrost and ice provide information concerning where and how should we search for evidence of life elsewhere in the Cosmos. The permafrost and ice wedges of the Fox tunnel preserves a magnificent of record of Pliocene, Pleistocene and Holocene life on Earth spanning more than 2.5 million years. This record includes frozen fossil bacteria, archaea, algae, mosses, higher plants, insects and mammals. In this paper we present the preliminary results of studies of the morphology, ultramicrostructure and elemental distributions of Fox tunnel microbiota as determined in-situ by the Environmental Scanning Electron Microscope (ESEM) and the Field Emission Scanning Electron Microscope (FESEM) investigations. The long-term viability of cryopreserved microbiota and potential implications to Astrobiology will be discussed.

  14. Cell viability and angiogenic potential of a bioartificial adipose substitute.

    Science.gov (United States)

    Panneerselvan, Anitha; Nguyen, Luong T H; Su, Yan; Teo, Wee Eong; Liao, Susan; Ramakrishna, Seeram; Chan, Ching Wan

    2015-06-01

    An implantable scaffold pre-seeded with cells needs to remain viable and encourage rapid angiogenesis in order to replace injured tissues, especially for tissue defect repairs. We created a bioartificial adipose graft composed of an electrospun 3D nanofibrous scaffold and fat tissue excised from New Zealand white rabbits. Cell viability and angiogenesis potential of the bioartificial substitute were examined during four weeks of culture in Dulbecco's Modified Eagle Medium by immunohistochemical staining with LIVE/DEAD® cell kit and PECAM-1 antibody, respectively. In addition, a Matrigel® assay was performed to examine the possibility of blood vessels sprouting from the bioartificial graft. Our results showed that cells within the graft were viable and vascular tubes were present at week 4, while cells in a fat tissue block were dead in vitro. In addition, capillaries were observed sprouting from the graft into the Matrigel, demonstrating its angiogenic potential. We expect that improved cell viability and angiogenesis in the bioartificial substitute, compared to intact autologous graft, could potentially contribute to its survival following implantation. Copyright © 2012 John Wiley & Sons, Ltd.

  15. No. 347-Obstetric Management at Borderline Viability.

    Science.gov (United States)

    Ladhani, Noor Niyar N; Chari, Radha S; Dunn, Michael S; Jones, Griffith; Shah, Prakesh; Barrett, Jon F R

    2017-09-01

    The primary objective of this guideline was to develop consensus statements to guide clinical practice and recommendations for obstetric management of a pregnancy at borderline viability, currently defined as prior to 25+6 weeks. Clinicians involved in the obstetric management of women whose fetus is at the borderline of viability. Women presenting for possible birth at borderline viability. This document presents a summary of the literature and a general consensus on the management of pregnancies at borderline viability, including maternal transfer and consultation, administration of antenatal corticosteroids and magnesium sulfate, fetal heart rate monitoring, and considerations in mode of delivery. Medline, EMBASE, and Cochrane databases were searched using the following keywords: extreme prematurity, borderline viability, preterm, pregnancy, antenatal corticosteroids, mode of delivery. The results were then studied, and relevant articles were reviewed. The references of the reviewed studies were also searched, as were documents citing pertinent studies. The evidence was then presented at a consensus meeting, and statements were developed. The content and recommendations were developed by the consensus group from the fields of Maternal-Fetal Medicine, Neonatology, Perinatal Nursing, Patient Advocacy, and Ethics. The quality of evidence was rated using criteria described in the Grading of Recommendations Assessment, Development and Evaluation methodology framework (reference 1). The Board of the Society of Obstetricians and Gynaecologists of Canada approved the final draft for publication. The quality of evidence was rated using the criteria described in the Grading of Recommendations, Assessment, Development, and Evaluation methodology framework. The interpretation of strong and weak recommendations is described later. The Summary of Findings is available upon request. A multidisciplinary approach should be used in counselling women and families at borderline

  16. Textural characteristics of fresh and frozen meat chicken and turkey

    Directory of Open Access Journals (Sweden)

    M. M. Momchilovа

    2016-01-01

    Full Text Available The purpose of the study is to determine the structural characteristics of the texture of chilled and frozen meat of chickens and turkeys. The object of the study were chicken and turkey breast and legs frozen and refrigerated. Research was carried out using two methods - the analysis of the profile of texture (TPA and shear Warner-Bratzler (WB. The total chemical composition determined by methods: total protein - Kjeldahl, carbohydrates, fats and total ash down in the relevant known methods. Samples have similar protein containing identical anatomical sites, have some differences in the content of fat and minerals. The results show that the water content of the turkey breasts and legs lower than with the chicken at about equal protein content. According to analysis of variance (ANOVA, a statistical difference was observed between the shear forces of refrigerated and frozen chicken legs, but no significant differences in the results of the shear characteristics of the refrigerated and frozen chicken breast samples were detected. Parameters produced by Warner-Bratzler correlates well with the characteristics of strength, hardness and wiry, obtained by texture profile analysis. The results indicate that the test samples have different textural characteristics depending on the type of meat and the thermal state. Methods of Warner-Bratzler and ATP method in combination with others, may be used to determine in which the thermal state (chilled or frozen were meat turkey and chicken.

  17. Viability of encapsulated Lactobacillus sp. Mar 8

    Directory of Open Access Journals (Sweden)

    EVI TRIANA

    2006-04-01

    Full Text Available Lactobacillus sp. Mar 8 had advantages as probiotic digestive system cholesterol lowering Lactobacillus. Applying in industry, particular processing technique is necessary for gaining product that ready for marketing and consuming. Spray drying is common technique using in various food processing. High processing temperature, 100-200oC, for 3-10 second become the barrier because cells were under extreme temperature stress. Therefore, encapsulate was needed to protect the cells from those extreme conditions. Viability and survival rate of encapsulated Lactobacillus sp. Mar 8 have been investigated. The result showed that Lactobacillus sp. Mar 8 that was encapsulated by 10% skim milk has higher viability than those by 5% skim milk, namely 72.37% and 51.69% respectively. Survival rate of encapsulated Lactobacillus cells will come to zero in 41.28 years. Therefore, encapsulated Lactobacillus sp. Mar 8 may use as probiotic agent.

  18. Twin pregnancy obtained with frozen-thawed embryos after in vitro maturation in a patient with polycystic ovarian syndrome

    OpenAIRE

    Godin, P. A.; Gaspard, Olivier; Thonon, Fabienne; Jouan, Caroline; Wijzen, F.; Dubois, Michel; Foidart, Jean-Michel

    2003-01-01

    Purpose: A twin pregnancy was obtained in a patient with polycystic ovary syndrome after the transfer of three in vitro maturation-derived day 3 embryos that has been frozen and thawed. Methods: The patient had received mild hMG stimulation followed by hCG injection. After culture for 24 - 48 h, mature oocytes were fertilized by ICSI. Embryos were cultured until day 3; supernumerary embryos were cryopreserved using a slow protocol. Results: Among 15 nonatretic oocytes, 9 matured, 8 were ferti...

  19. Influence of electrospun scaffolds prepared from distinct polymers on proliferation and viability of endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Matveeva, V. G., E-mail: matveeva-vg@mail.ru; Antonova, L. V., E-mail: antonova.la@mail.ru; Velikanova, E. A.; Sergeeva, E. A.; Krivkina, E. O.; Glushkova, T. V.; Kudryavtseva, Yu. A.; Barbarash, O. L.; Barbarash, L. S. [Research Institute for Complex Issues of Cardiovascular Diseases, Kemerovo, 650002 (Russian Federation)

    2015-10-27

    We compared electrospun nonwoven scaffolds from polylactic acid (PLA), polycaprolactone (PCL), and polyhydroxybutyrate/valerate (PHBV)/polycaprolactone (PHBV/PCL). The surface of PHBV/PCL and PCL scaffolds was highly porous and consisted of randomly distributed fibers, whilst the surface of PLA scaffolds consisted of thin straight fibers, which located more sparsely, forming large pores. Culture of EA.hy 926 endothelial cells on these scaffolds during 7 days and further fluorescent microscopy demonstrated that the surface of PHBV/PCL scaffolds was most favorable for efficient adhesion, proliferation, and viability of endothelial cells. The lowest proliferation rate and cell viability were detected on PLA scaffolds. Therefore, PHBV/PCL electrospun nonwoven scaffolds demonstrated the best results regarding endothelial cell proliferation and viability as compared to PCL and PLA scaffolds.

  20. Stiffness Evolution in Frozen Sands Subjected to Stress Changes

    KAUST Repository

    Dai, Sheng

    2017-04-21

    Sampling affects all soils, including frozen soils and hydrate-bearing sediments. The authors monitor the stiffness evolution of frozen sands subjected to various temperature and stress conditions using an oedometer cell instrumented with P-wave transducers. Experimental results show the stress-dependent stiffness of freshly remolded sands, the dominant stiffening effect of ice, creep after unloading, and the associated exponential decrease in stiffness with time. The characteristic time for stiffness loss during creep is of the order of tens of minutes; therefore it is inevitable that frozen soils experience sampling disturbances attributable to unloading. Slow unloading minimizes stiffness loss; conversely, fast unloading causes a pronounced reduction in stiffness probably attributable to the brittle failure of ice or ice-mineral bonding.

  1. Fluorescent antibody-viability staining and beta-glucuronidase assay as rapid methods for monitoring Escherichia coli viability in coastal marine waters.

    Science.gov (United States)

    Caruso, G; De Pasquale, F; Mancuso, M; Zampino, D; Crisafi, E

    2006-01-01

    A faecal pollution monitoring of coastal Messina waters was performed by comparing three (microscopic, enzyme, and culture) methods. Evidence of Escherichia coli cells (29.99 to 96.79% of the total enteropathogenic serotypes) retaining their viability into the marine environment was shown. beta-Glucuronidase activity rates suggested that living cells were also metabolically active. Heavily polluted sites were detected, where improperly treated urban wastes were discharged. Significant relationships between microscopic and enzymatic data proved both methods to be suitable alternatives to the culture method for E. coli detection, improving environmental quality assessment.

  2. Lactose-free frozen yogurt: production and characteristics.

    Science.gov (United States)

    Skryplonek, Katarzyna; Gomes, David; Viegas, Jorge; Pereira, Carlos; Henriques, Marta

    2017-01-01

    Nowadays, consumer demand is driving better and more nutritious dairy products. Changing from traditional to new lactose-free products poses technological challenges for the food industry in order to maintain or improve their food characteristics and consumer preferences. This study investigates the production of lactose-free frozen yogurt by enzymati- cally hydrolysis of lactose, and its influence on the final product characteristics. In the case of lactose-free products, commercial Ha-lactase® was used for hydrolysis, and the reaction occurred simultaneously with fermentation. The effect of lactose hydrolysis on the physicochemical properties, texture, viscosity, overrun and sensory attributes in the final product was investigated. After yogurt maturation, the acidity of the lactose-free product was significantly higher than in the control, suggesting that breaking down lactose enhances the fermentation process. Lactose-free frozen yogurt had significantly lower hardness and stickiness and higher viscosity than control frozen yogurt. Moreover, lactose hydrolysis promoted a smooth and creamy consistency, whereas in the case of conventional prod- ucts a coarse structure, due to the presence of large ice crystals, was identified. Hydrolysis of lactose also improved the sweetness and brightness of frozen yogurt. The improved textural properties of lactose-free product results from the fact that monosaccharides produced during lactose hydrolysis depress the freezing point of the mix, which enables product with softer structure and bigger resistance to ice recrystallization to be obtained. The study showed that lactose-free frozen yogurt may be used successfully for production of novel lactose-free frozen desserts. Lactose hydrolysis improves the texture and viscosity of the product,     as well as enhancing its sensory quality.

  3. Segmental infarction of the testis: can frozen sections avoid orchidectomy?

    Science.gov (United States)

    Pacella, E; Grillo, F; Lapetina, C; Cabiddu, F; Auriati, L; Tunesi, G; Mastracci, L

    2015-03-01

    Segmental infarction (SI) of the testis is a rare condition that can masquerade as a mass lesion, thus requiring exclusion of tumour. If clinical exams do not exclude a neoplastic lesion with certainty, orchidectomy is usually performed. A case of SI of the testis is presented; the use of frozen section of the enucleated mass demonstrated the ischaemic nature of the lesion, so avoiding orchidectomy. The 8 year follow-up was uneventful. The use of frozen section in SI could permit the selection of cases in which testicular-sparing surgery should be considered.

  4. Successful birth of the first frozen oocyte baby in India

    Directory of Open Access Journals (Sweden)

    Priya Selvaraj

    2009-01-01

    Full Text Available We report the first pregnancy and birth in India after the transfer of embryos generated from frozen- thawed oocytes. A 29-year-old woman with previous bad obstetric history and an abnormal karyotype, necessitating donor oocyte programme. Embryos were generated by microinjection of frozen-thawed sperms into thawed human oocytes (intracytoplasmic sperm injection. This resulted in an healthy male baby with a birth weight of 2.54 kg which was born by cesarean section at 35-36 weeks of gestation with normal follow-up. Thus oocyte cryopreservation can be performed with reproducible success leading to a viable offspring.

  5. Cryoelectrolysis—electrolytic processes in a frozen physiological saline medium

    Science.gov (United States)

    Lugnani, Franco; Macchioro, Matteo

    2017-01-01

    Background Cryoelectrolysis is a new minimally invasive tissue ablation surgical technique that combines the ablation techniques of electrolytic ablation with cryosurgery. The goal of this study is to examine the hypothesis that electrolysis can take place in a frozen aqueous saline solution. Method To examine the hypothesis we performed a cryoelectrolytic ablation protocol in which electrolysis and cryosurgery are delivered simultaneously in a tissue simulant made of physiological saline gel with a pH dye. We measured current flow, voltage and extents of freezing and pH dye staining. Results Using optical measurements and measurements of currents, we have shown that electrolysis can occur in frozen physiological saline, at high subzero freezing temperatures, above the eutectic temperature of the frozen salt solution. It was observed that electrolysis occurs when the tissue resides at high subzero temperatures during the freezing stage and essentially throughout the entire thawing stage. We also found that during thawing, the frozen lesion temperature raises rapidly to high subfreezing values and remains at those values throughout the thawing stage. Substantial electrolysis occurs during the thawing stage. Another interesting finding is that electro-osmotic flows affect the process of cryoelectrolysis at the anode and cathode, in different ways. Discussion The results showing that electrical current flow and electrolysis occur in frozen saline solutions imply a mechanism involving ionic movement in the fluid concentrated saline solution channels between ice crystals, at high subfreezing temperatures. Temperatures higher than the eutectic are required for the brine to be fluid. The particular pattern of temperature and electrical currents during the thawing stage of frozen tissue, can be explained by the large amounts of energy that must be removed at the outer edge of the frozen lesion because of the solid/liquid phase transformation on that interface. Conclusion

  6. Effect of storage temperature on the viability of human periodontal ligament fibroblasts.

    Science.gov (United States)

    Chen, Fubo; Qi, Shengcai; Lu, Liyan; Xu, Yuanzhi

    2015-02-01

    The viability of periodontal ligament fibroblasts (PDLF) can affect the long-term prognosis of replanted avulsed teeth. When immediate replantation of an avulsed tooth is not possible, the cells should be incubated in a physiological storage medium instantly to maintain their biological activity. The ability of different storage media to preserve PDLF viability has been previously evaluated. However, few studies have showed the effect of temperature on the viability of PDLF cultured with various storage media in vitro. This study was designed to measure PDLF activity by CCK-8 assay to compare the effectiveness at 4, 22 (room temperature), and 37°C under various storage media. Statistical analysis demonstrated that tap water, saline, and saliva decreased cell viability as the storage temperature increased. But the temperature played only a minor role on cell viability when cells were incubated in Hank's balanced salt solution (HBSS), Dubelco's modified Eagle's medium (DMEM), or milk. Within the parameters of this study, it seems that room temperature is adequate for storing the avulsed teeth in HBSS, DMEM, or milk in the extra-alveolar period. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  7. Electrotherapy modalities for adhesive capsulitis (frozen shoulder).

    Science.gov (United States)

    Page, Matthew J; Green, Sally; Kramer, Sharon; Johnston, Renea V; McBain, Brodwen; Buchbinder, Rachelle

    2014-10-01

    Adhesive capsulitis (also termed frozen shoulder) is a common condition characterised by spontaneous onset of pain, progressive restriction of movement of the shoulder and disability that restricts activities of daily living, work and leisure. Electrotherapy modalities, which aim to reduce pain and improve function via an increase in energy (electrical, sound, light, thermal) into the body, are often delivered as components of a physical therapy intervention. This review is one in a series of reviews which form an update of the Cochrane review 'Physiotherapy interventions for shoulder pain'. To synthesise the available evidence regarding the benefits and harms of electrotherapy modalities, delivered alone or in combination with other interventions, for the treatment of adhesive capsulitis. We searched CENTRAL, MEDLINE, EMBASE, CINAHL Plus and the ClinicalTrials.gov and World Health Organization (WHO) International Clinical Trials Registry Platform (ICTRP) clinical trials registries up to May 2014, unrestricted by language, and reviewed the reference lists of review articles and retrieved trials to identify any other potentially relevant trials. We included randomised controlled trials (RCTs) and controlled clinical trials using a quasi-randomised method of allocation that included adults with adhesive capsulitis and compared any electrotherapy modality to placebo, no treatment, a different electrotherapy modality, or any other intervention. The two main questions of the review focused on whether electrotherapy modalities are effective compared to placebo or no treatment, or if they are an effective adjunct to manual therapy or exercise (or both). The main outcomes of interest were participant-reported pain relief of 30% or greater, overall pain, function, global assessment of treatment success, active shoulder abduction, quality of life, and the number of participants experiencing any adverse event. Two review authors independently selected trials for inclusion

  8. Modeling the impact of high temperatures on microalgal viability and photosynthetic activity.

    Science.gov (United States)

    Béchet, Quentin; Laviale, Martin; Arsapin, Nicolas; Bonnefond, Hubert; Bernard, Olivier

    2017-01-01

    Culture collapse due to high temperatures can significantly impact the profitability of outdoor algal cultivation systems. The objective of this study was to model for the first time the impact of high temperatures on algal activity and viability. Viability measurements on Dunaliella salina cultures were based on cytometry with two fluorescent markers (erythrosine and fluorescein di-acetate), and photosynthetic activity was measured by Pulse Amplitude Modulation (PAM) fluorometry. Kinetic studies revealed that viability and activity losses during exposure to high temperatures could be described by a Weibull model. Both mortality and activity were shown to be functions of the thermal dose received by the algae, defined as the product of duration of exposure to high temperatures and an exponential function of temperature. Simulations at five climatic locations revealed that culture collapse due to high temperatures could impact productivity of D. salina in non-temperature-controlled outdoor photobioreactors by 35 and 40% in arid and Mediterranean climates, respectively. The model developed in this study can be used to forecast the impact of high temperatures on algal biofuel productivity. When coupled with models predicting the temperature of outdoor cultivation systems, this model can also be used to select the best combination of location, system geometry, and algal species to minimize the risks of culture collapse and therefore maximize biofuel productivity.

  9. Combined Effect of Trolox and EDTA on Frozen-Thawed Sperm Quality

    Directory of Open Access Journals (Sweden)

    Sara Keshtgar

    2016-05-01

    Full Text Available The freezing and thawing process not only is associated with serious damage to sperm such as damage to the plasma membrane and the acrosomal membrane but also changes the membrane permeability to some ions including calcium. Also, the generation of oxygen free radicals is increased during the freezing-thawing process. The purpose of this study was to evaluate of the effects of Trolox as an antioxidant and edetic acid (EDTA as a calcium chelator on frozen-thawed (FT sperm and compare these effects with those on fresh sperm. This study was done on these men of 25 healthy men, who referred to Shiraz Infertility Centerbetween2012 and2013. Normal samples were transferred to the ReproductivePhysiology Laboratory, Department of Physiology,Shiraz University of Medical Sciences, Shiraz. The samples were divided into two groups randomly: fresh and FT sperm groups. Each group was divided into five subgroups: control group, the solvent group (0.1%dimethyl sulfoxide [DMSO], Trolox group (200μM, EDTA group (1.1mM, and Trolox+EDTA group. The percentages of motility, viability, and acrosome-reacted sperm were tested. The percentages of motility and viability in the FT sperm were lower than those in the fresh sperm. The progressive motility of the FT sperm was improved nonsignificantly with Trolox+EDTA. However, the effect of Trolox+EDTA on the progressive motility of the FT sperm was much more than that on the fresh sperm. The fewest acrosome-reacted sperm were observed in the EDTA-containingFT sperm. Antioxidant supplementation or omission of extracellular calcium may partly improve motility and also reduce acrosomal damage in FT sperm.

  10. Combined Effect of Trolox and EDTA on Frozen-Thawed Sperm Quality.

    Science.gov (United States)

    Keshtgar, Sara; Iravanpour, Farideh; Gharesi-Fard, Behrooz; Kazerooni, Marjaneh

    2016-05-01

    The freezing and thawing process not only is associated with serious damage to sperm such as damage to the plasma membrane and the acrosomal membrane but also changes the membrane permeability to some ions including calcium. Also, the generation of oxygen free radicals is increased during the freezing-thawing process. The purpose of this study was to evaluate of the effects of Trolox as an antioxidant and edetic acid (EDTA) as a calcium chelator on frozen-thawed (FT) sperm and compare these effects with those on fresh sperm. This study was done on these men of 25 healthy men, who referred to Shiraz Infertility Centerbetween2012 and2013. Normal samples were transferred to the ReproductivePhysiology Laboratory, Department of Physiology, Shiraz University of Medical Sciences, Shiraz. The samples were divided into two groups randomly: fresh and FT sperm groups. Each group was divided into five subgroups: control group, the solvent group (0.1%dimethyl sulfoxide [DMSO]), Trolox group (200μM), EDTA group (1.1mM), and Trolox+EDTA group. The percentages of motility, viability, and acrosome-reacted sperm were tested. The percentages of motility and viability in the FT sperm were lower than those in the fresh sperm. The progressive motility of the FT sperm was improved nonsignificantly with Trolox+EDTA. However, the effect of Trolox+EDTA on the progressive motility of the FT sperm was much more than that on the fresh sperm. The fewest acrosome-reacted sperm were observed in the EDTA-containingFT sperm. Antioxidant supplementation or omission of extracellular calcium may partly improve motility and also reduce acrosomal damage in FT sperm.

  11. Mammalian Cell Tissue Culture Techniques.

    Science.gov (United States)

    Phelan, Katy; May, Kristin M

    2016-06-01

    Cultured tissues and cells are used extensively in physiological and pharmacological studies. In vitro cultures provide a means of examining cells and tissues without the complex interactions that would be present if the whole organism were studied. A number of special skills are required in order to preserve the structure, function, behavior, and biology of cells in culture. This unit describes the basic skills required to maintain and preserve cell cultures: maintaining aseptic technique, preparing media with the appropriate characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

  12. Rapid onsite assessment of spore viability.

    Energy Technology Data Exchange (ETDEWEB)

    Branda, Steven; Lane, Todd W.; VanderNoot, Victoria A.; Gaucher, Sara P.; Jokerst, Amanda S.

    2005-12-01

    This one year LDRD addresses problems of threat assessment and restoration of facilities following a bioterror incident like the incident that closed down mail facilities in late 2001. Facilities that are contaminated with pathogenic spores such as B. anthracis spores must be shut down while they are treated with a sporicidal agent and the effectiveness of the treatment is ascertained. This process involves measuring the viability of spore test strips, laid out in a grid throughout the facility; the CDC accepted methodologies require transporting the samples to a laboratory and carrying out a 48 hr outgrowth experiment. We proposed developing a technique that will ultimately lead to a fieldable microfluidic device that can rapidly assess (ideally less than 30 min) spore viability and effectiveness of sporicidal treatment, returning facilities to use in hours not days. The proposed method will determine viability of spores by detecting early protein synthesis after chemical germination. During this year, we established the feasibility of this approach and gathered preliminary results that should fuel a future more comprehensive effort. Such a proposal is currently under review with the NIH. Proteomic signatures of Bacillus spores and vegetative cells were assessed by both slab gel electrophoresis as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection. The conditions for germination using a number of chemical germinants were evaluated and optimized and the time course of protein synthesis was ascertained. Microseparations were carried out using both viable spores and spores inactivated by two different methods. A select number of the early synthesis proteins were digested into peptides for analysis by mass spectrometry.

  13. Influence of storage temperature on growth of Penicillium polonicum and Penicillium glabrum and potential for deterioration of frozen chicken nuggets.

    Science.gov (United States)

    Saccomori, Fernanda; Wigmann, Évelin Francine; Bernardi, Angélica Olivier; Alcano-González, María de Jesús; Copetti, Marina Venturini

    2015-05-04

    The practice of freezing food is one of the main processes used by the industry to prolong the shelf life of foods. Its use has expanded in recent years due to the increased consumption of convenience products, many of which are sold in frozen form. The temperature at which these foods are maintained during marketing in supermarkets or stored in the consumer's home is critical to ensure microbiological stability of products. Temperature abuse can allow microbial growth, especially growth of filamentous psychrophilic fungi. Besides economic losses in the industrial sector due to the return of products and loss of confidence by consumers, the development of fungi in foods is a public health problem due to the possibility of mycotoxin production. The aim of this study was to assess the growth at temperatures of 5, 0, -5 and -18°C for two species of fungi involved in the deterioration of frozen chicken nuggets, Penicillium polonicum (33/12 NGT) and Penicillium glabrum (29/12 NGT), inoculated both in culture medium and in the food. The results demonstrated that P. polonicum was able to form microcolonies on potato dextrose agar plates at 0°C and form visible colonies on the surface of the frozen chicken nuggets kept at -5°C for 120 days, regardless of brand. For P. glabrum the limiting growth temperature was 5°C in the culture medium and 0°C on frozen chicken nuggets, regardless of the brand analyzed. Thus, it is essential to adhere to the storage temperatures recommended to ensure the stability and safety of this food product. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Cryoprotectant redistribution along the frozen straw probed by Raman spectroscopy.

    Science.gov (United States)

    Karpegina, Yu A; Okotrub, K A; Brusentsev, E Yu; Amstislavsky, S Ya; Surovtsev, N V

    2016-04-01

    The distribution of cryoprotectant (10% glycerol) and ice along the frozen plastic straw (the most useful container for freezing mammalian semen, oocytes and embryos) was studied by Raman scattering technique. Raman spectroscopy being a contactless, non-invasive tool was applied for the straws filled with the cryoprotectant solution and frozen by controlled rate programs commonly used for mammalian embryos freezing. Analysis of Raman spectra measured at different points along the straw reveals a non-uniform distribution of the cryoprotectant. The ratio between non-crystalline solution and ice was found to be increased by several times at the bottom side of the solution column frozen by the standard freezing program. The increase of the cryoprotectant fraction occurs in the area where embryos or oocytes are normally placed during their freezing. Possible effects of the cooling rate and the ice nucleation temperature on the cryoprotectant fraction at the bottom side of the solution column were considered. Our findings highlight that the ice fraction around cryopreserved embryos or oocytes can differ significantly from the averaged one in the frozen plastic straws. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Plastic Straws for Frozen Semen (A Field Evaluation)

    Science.gov (United States)

    Macpherson, J. W.; King, G. J.

    1966-01-01

    A field trial was conducted to compare the routine frozen semen methods presently used at this laboratory with those reported by French workers. When the extender used was sterile milk both methods resulted in fertility equal to the standard. Fertility results obtained with the French method employing Lactiphos extender were significantly lower than the standard. ImagesFig. 1. PMID:4225285

  16. Plastic straws for frozen semen. (A field evaluation).

    Science.gov (United States)

    Macpherson, J W; King, G J

    1966-04-01

    A field trial was conducted to compare the routine frozen semen methods presently used at this laboratory with those reported by French workers. When the extender used was sterile milk both methods resulted in fertility equal to the standard. Fertility results obtained with the French method employing Lactiphos extender were significantly lower than the standard.

  17. Characteristics of frozen colostrum thawed in a microwave oven

    Energy Technology Data Exchange (ETDEWEB)

    Jones, L.R.; Taylor, A.W.; Hines, H.C.

    1987-09-01

    Use of a microwave oven to thaw frozen colostrum was evaluated. Colostrum was collected from nine cows, four of which were immunized to produce specific colostral antibodies. Colostrum from each cow was frozen, subsequently thawed, and pooled. One-liter aliquots of the pooled colostrum were frozen and assigned randomly to three thawing treatments. Colostrum was thawed using one of three regimens: 10 min in a microwave oven at full power (650 W), 17 min in a microwave oven at half power (325 W), and 25 min in 45 degrees C water. Colostrum thawed in the microwave oven was slightly coagulated and had lower volume and total protein content than colostrum thawed in water. Casein and pH were not different among treatments. Both concentration and total content of immunoglobulin A were higher in the control than in microwave treatments. Neither amount nor concentration of immunoglobulin G and immunoglobulin M were different among treatments. Immunological activity, measured by a hemolytic test, was lower for microwave treatments than the control but did not differ between microwave treatments. Frozen colostrum thawed in a microwave oven should provide a reasonable source of colostrum when fresh high quality colostrum is not available.

  18. Parasites and Morphometric Indices of Frozen Fish Sold in Nsukka ...

    African Journals Online (AJOL)

    The parasites and morphometric indices of frozen fish sold in Nsukka Urban Market in Nsukka Local Government Area of Enugu State, Nigeria were investigated between June and December 2008 to determine the parasite prevalence, mean intensity and abundance and some morphometric indices associated with ...

  19. Frozen shoulder or missed posterior dislocation? | Leijnen | South ...

    African Journals Online (AJOL)

    The clinical presentation of a missed posterior shoulder dislocation has similarities with an idiopathic frozen shoulder masking proper diagnosis at the time of injury. We report a case of a 48-year-old woman with shoulder pain which demonstrates the importance of correct initial diagnosis and management.

  20. Monitoring and analysis of frozen debris lobes, phase IB.

    Science.gov (United States)

    2015-09-01

    Frozen debris lobes (FDLs) are slow-moving landslides in permafrost, many of which are present within the Dalton Highway corridor in the Brooks Range of Alaska. During this phase of the research, we continued our investigations of FDL-A (the closest ...

  1. Development of frozen-fried yam slices: Optimization of the ...

    African Journals Online (AJOL)

    The industrial process of production of frozen French fries traditionally includes a pre-frying step. ... An adiabatic system was also developed by means of an insulator in which the core temperature of fried yam slices can be maintained constant at about 55°C after 15min of cooling, facilitating texture measurements at ...

  2. Smoked And frozen fish consumption and marketing channels in the ...

    African Journals Online (AJOL)

    Eighty-six per cent (86%) of consumers preferred smoked fish while 14% preferred frozen fish. Herrings, horse mackerel and salmon were the most preferred fish. Consumer preferences were based on taste, affordability, availability and wholesomeness in soup and stew. The fishmongers indicated that they obtained their ...

  3. DYNAMIC LINKAGES BETWEEN PRICES AND IMPORTS FOR JAPANESE FROZEN TUNA

    OpenAIRE

    Lei, Li-Fen; Sun, Lih-Chyun; Liu, His-Chiang

    1998-01-01

    VAR models have been used to describe the dynamic relationships among market price, Japanese harvest, import-from-Taiwan, and import-from-Korea for frozen Big-Eye tuna and Yellow-Fin tuna markets in Japan. It is found that tuna imports from South Korea exert more significant effects on Japan market prices than import-from-Taiwan.

  4. The effects of commercial fibres on frozen bread dough

    Directory of Open Access Journals (Sweden)

    JELENA FILIPOVIĆ

    2010-02-01

    Full Text Available The daily intake of dietary fibres in highly industrialized countries is at a low level and, therefore, adversely affecting human health. The objective of this research was to analyze the influence of different commercial fibres (originating from sugar beet pulp fibrex, and Jerusalem artichoke inulin HPX and GR in yeast dough at a level of 5 %, on the rheological properties of dough and the quality of bread during frozen storage. Frozen dough characteristics were determined using a Brabender maturograph and test baking was followed according the AACC procedure. The dough was frozen at –18 °C and stored over a period of 60 days. The results concerning the dough (proving time and stability and bread quality (volume and crumb quality were statistically analyzed by multivariance Manova and discriminative analysis, which indicated that there was a significant difference between dough without fibres and dough with different fibres (fibrex, inulin HPX and GR. The discrimination coefficient points that the greatest influence of fibres on the final proof and proving stability is after 30 days (6.250 and after 0 days (6.158, respectively, but the greatest influence of fibres on bread volume and bread crumb quality (15.488 and 3.638, respectively can be expected on non frozen dough, due to above mention their adverse the effect on gluten network.

  5. The Erosion of Frozen Argon by Swift Helium Ions

    DEFF Research Database (Denmark)

    Besenbacher, F.; Bøttiger, Jørgen; Graversen, O.

    1981-01-01

    The temperature, energy, and thickness dependence of the erosion rates of frozen argon films when irradiated with 0.1–3 MeV helium ions have been measured. The erosion yields Y are much too high to be explained by the concentional collisional cascade-sputtering theory and are furthermore...

  6. coagulation factors level in fresh frozen plasma in rwanda

    African Journals Online (AJOL)

    2014-02-02

    Feb 2, 2014 ... compared with the calculated value to get the effect of storage conditions (time and ... month up to three months of storage had been stable. Plot of coagulation factors levels in fresh frozen based on time point and sex .... reversal of therapy with vitamin K antagonists(8). Especially in emergency situations, ...

  7. 21 CFR 146.120 - Frozen concentrate for lemonade.

    Science.gov (United States)

    2010-04-01

    ... 146.120 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... lemonade is the frozen food prepared from one or both of the lemon juice ingredients specified in paragraph... percent by weight. (b) The lemon juice ingredients referred to in paragraph (a) of this section are: (1...

  8. Cryo-scanning electron microscopy discloses differences in dehydration of frozen boar semen stored in large containers.

    Science.gov (United States)

    Ekwall, H

    2009-02-01

    In general, freezing in flat plastic polyethylene terephthalate (PET) bags (FlatPacks) at 50 degrees C/min gives better post-thaw viability, in terms of sperm motility and membrane integrity, than does freezing in plastic maxi-straws, probably owing to differences in cryobiology. To test the hypothesis that this better survival post-thaw relates to the degree of sperm dehydration during freezing, the present study investigated the structure of boar semen in a frozen state using cryo-scanning electron microscopy (cryo-SEM) to compare two different packages (FlatPacks and maxi-straws) for single artificial insemination (AI) doses, and three different freezing rates. The semen was split-sample frozen in maxi-straws or FlatPacks (both holding 5 ml) using 3% glycerol as cryoprotectant. Three freezing rates were applied from -5 degrees C to -100 degrees C, namely 2 degrees C/min, 50 degrees C/min and 1200 degrees C/min, the lattermost by plunging the samples into liquid nitrogen (LN(2)). The samples were thereafter fractured into LN(2) and larger areas of extra-cellular, unbound frozen water ('ice lakes') were measured to determine the degree of dehydration of the spermatozoa. These areas decreased in size with an increase in cooling rate, the differences in size being more dramatic for maxi-straws than for FlatPacks. Size of ice lakes was also influenced by location within package in relation to cooling rate, the central values being always smaller in maxi-straws than in Flatpacks (p < 0.05 at 2 degrees C/min and 50 degrees C/min) but not at 1200 degrees C/min, which suggested the FlatPack allows for more homogenous freezing of boar semen.

  9. SENSORY AND CHEMICAL ASPECTS OF FROZEN SOY YOGURT FERMENTED WITH Enterococcus faecium AND Lactobacillus jugurti

    Directory of Open Access Journals (Sweden)

    D. P. MIGUEL

    2008-09-01

    Full Text Available

    As recent studies have demonstrated the beneficial effects to health of soy yogurt fermented with E. faecium and L. jugurti, aim to offer new options of consumption or this product, the goal of this work was to present the processing of frozen soy yogurt and the evaluation of its sensory characteristics. Then compare it with the other ice creams regularly processed with cow and soymilk fermented with a mixed culture of L. delbrueckii ssp. bulgaricus and S. thermophilus and also with the non-fermented ice creams acidified with the addition of lactic acid. The effect of the lactic acid bacteria in the oxidation of the product was also evaluated. The results demonstrated that it is possible to have a frozen soy yogurt fermented with E. faecium and L. jugurti with good sensory characteristics up to a period of 180 days. The lactic bacteria were not able to stop the development of the oxidation process for a long period of storage but it did not alter the sensory characteristics of the product.

  10. Evaluation of Morphological Change and Aggregation Process of Ice Crystals in Frozen Food by Using Fractal Analysis

    Science.gov (United States)

    Koshiro, Yoko; Watanabe, Manabu; Takai, Rikuo; Hagiwara, Tomoaki; Suzuki, Toru

    Size and shape of ice crystals in frozen food materials are very important because they affect not only quality of foods but also the viability of industrial processing such as freeze-drying of concentration. In this study, 30%wt sucrose solution is used as test samples. For examining the effect of stabilizerspectine and xantan gum is added to the sucrose solution. They are frozen on the cold stage of microscope to be observed their growing ice crystals under the circumstance of -10°C. Their size and shape are measured and quantitatively evaluated by applying fractal analysis. lce crystal of complicated shape has large fractal dimension, and vice versa. It successflly categorized the ice crystals into two groups; one is a group of large size and complicated shape, and the other is a group of small size and plain shape. The critical crystal size between the two groups is found to become larger with increasing holding time. It suggests a phenomenological model for metamorphoses process of ice crystals. Further, it is indicated that xantan gum is able to suppress the smoothing of ice crystals.

  11. Population Viability Analysis of Riverine Fishes

    Energy Technology Data Exchange (ETDEWEB)

    Bates, P.; Chandler, J.; Jager, H.I.; Lepla, K.; Van Winkle, W.

    1999-04-12

    Many utilities face conflkts between two goals: cost-efficient hydropower generation and protecting riverine fishes. Research to develop ecological simulation tools that can evaluate alternative mitigation strategies in terms of their benefits to fish populations is vital to informed decision-making. In this paper, we describe our approach to population viability analysis of riverine fishes in general and Snake River white sturgeon in particular. We are finding that the individual-based modeling approach used in previous in-stream flow applications is well suited to addressing questions about the viability of species of concern for several reasons. Chief among these are: (1) the abiIity to represent the effects of individual variation in life history characteristics on predicted population viabili~, (2) the flexibili~ needed to quanti~ the ecological benefits of alternative flow management options by representing spatial and temporal variation in flow and temperaturty and (3) the flexibility needed to quantifi the ecological benefits of non-flow related manipulations (i.e., passage, screening and hatchery supplementation).

  12. 77 FR 60675 - Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Initiation of Antidumping...

    Science.gov (United States)

    2012-10-04

    ... International Trade Administration Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Initiation of Antidumping Duty New Shipper Review AGENCY: Import Administration, International Trade... antidumping duty order on certain frozen fish fillets (``fish'') from the Socialist Republic of Vietnam...

  13. 77 FR 20356 - Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Extension of Preliminary...

    Science.gov (United States)

    2012-04-04

    ... International Trade Administration Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Extension... AGENCY: Import Administration, International Trade Administration, Department of Commerce. SUMMARY: The... administrative review, and aligned new shipper reviews, of certain frozen fish fillets (``fish fillets'') from...

  14. Prediction of chemical, physical and sensory data from process parameters for frozen cod using multivariate analysis

    DEFF Research Database (Denmark)

    Bechmann, Iben Ellegaard; Jensen, H.S.; Bøknæs, Niels

    1998-01-01

    Physical, chemical and sensory quality parameters were determined for 115 cod (Gadus morhua) samples stored under varying frozen storage conditions. Five different process parameters (period of frozen storage, frozen storage. temperature, place of catch, season for catching and state of rigor) we...... by ones and zeroes only. These results illustrate the application of multivariate analysis as an effective strategy for improving the quality of frozen fish products. (C) 1998 Society of Chemical Industry......Physical, chemical and sensory quality parameters were determined for 115 cod (Gadus morhua) samples stored under varying frozen storage conditions. Five different process parameters (period of frozen storage, frozen storage. temperature, place of catch, season for catching and state of rigor) were...... varied systematically at two levels. The data obtained were evaluated using the multivariate methods, principal component analysis (PCA) and partial least squares (PLS) regression. The PCA models were used to identify which process parameters were actually most important for the quality of the frozen cod...

  15. Evaluation of the ovarian reserve in women transplanted with frozen and thawed ovarian cortical tissue

    DEFF Research Database (Denmark)

    Greve, Tine; Schmidt, Kirsten Tryde; Kristensen, Stine Gry

    2012-01-01

    To investigate ovarian reserve and ovarian function in women transplanted with frozen/thawed ovarian tissue.......To investigate ovarian reserve and ovarian function in women transplanted with frozen/thawed ovarian tissue....

  16. Inactivation of Paragonimus westermani metacercariae in soy sauce-marinated and frozen freshwater crabs.

    Science.gov (United States)

    Kim, Tae Im; Oh, Se-Ra; Dai, Fuhong; Yang, Hyun-Jong; Ha, Sang-Do; Hong, Sung-Jong

    2017-03-01

    Soy sauce-marinated freshwater crabs (Eriocheir japonicus) are a source of human paragonimiasis. The viability of Paragonimus westermani metacercariae (PwMc) in marinated crabs was investigated in an experimental setting. The PwMc collected from freshwater crayfish were inoculated into freshwater crabs, which were then frozen or marinated in soy sauce. All PwMc in the freshwater crabs were inactivated after freezing for 48 h at -20 °C and after freezing for 12 h at -40 °C. After marinating for 32 days, the survival rate of PwMc in 5% NaCl soy sauce was 50%, in 7.5% NaCl soy sauce it was 33.3%, and in 10.0% NaCl soy sauce it was 31.3%. When marinated for 64 days, all PwMc were inactivated in all experimental groups. These results revealed that freezing and soy sauce marination were detrimental to the survival of PwMc in freshwater crabs. Specifically, freezing crabs for more than 48 h or soaking them in soy sauce containing at least 5.0% NaCl for 64 days can inactivate PwMc. These results can inform the production of the traditional Korean soy sauce-marinated freshwater crabs known as gejang.

  17. Influence of pH on the physicochemical and sensorial characteristics of strawberry frozen yogurt

    OpenAIRE

    Pereira, Gustavo das Graças; Universidade Federal de Lavras; Rafael, Leonardo Mesquita; Universidade Federal de Lavras; Gajo, Adriano Alvarenga; Universidade Federal de Lavras; Ramos, Thaís de Melo; Universidade Federal de Lavras; Pinto, Sandra Maria; Universidade Federal de Lavras; Resende, Jaime Vilela de; Universidade Federal de Lavras; Abreu, Luiz Ronaldo de; Universidade Federal de Lavras

    2012-01-01

    Frozen yogurt is a frozen fermented dessert which presents structural characteristics similar to ice cream and nutritional and sensorial properties which resemble to yogurt. The acidification of ice cream mix by means of lactic bacteria can influence the properties of frozen yogurt. For that reason, the present work aimed to evaluate the influence of pH on the physicochemical and sensorial characteristics of strawberry frozen yogurt. The formulation consisted of 6 % of milk fat, 10 % of milk ...

  18. Long-term storage of peripheral blood stem cells frozen and stored with a conventional liquid nitrogen technique compared with cells frozen and stored in a mechanical freezer.

    Science.gov (United States)

    McCullough, Jeffrey; Haley, Rebecca; Clay, Mary; Hubel, Allison; Lindgren, Bruce; Moroff, Gary

    2010-04-01

    Cryopreservation of hematopoietic progenitor cells using liquid nitrogen and controlled-rate freezing requires complex equipment and highly trained staff and is expensive. We compared the liquid nitrogen method with methods using a combination of dimethyl sulfoxide (DMSO) and hydroxyethyl starch (HES) for cryopreservation followed by storage in mechanical freezers. Peripheral blood stem cells (PBSCs) were collected from normal donors by apheresis and allocated to one of four preservation and storage conditions: 1) 10% DMSO with freezing in liquid nitrogen and storage in liquid nitrogen, 2) 5% DMSO and 6% HES with freezing and storage in a -80 degrees C mechanical freezer, 3) 5% DMSO and 6% HES with freezing in a -80 degrees C mechanical freezer and storage in a -135 degrees C mechanical freezer, or 4) 5% DMSO and 6% HES with freezing and storage both in a 135 degrees C mechanical freezer. Cells were stored for 5 years during which total nucleated cells (TNCs), cell viability, CD34+ cell content, and colony-forming unit-granulocyte-macrophage content were determined. There were some significant differences in the variables measured during freezing and the 5 years of storage compared to the values before freezing and storage; however, these differences were not consistent and do not favor one protocol over the others. Samples stored for 24 hours before cryopreservation showed a significant decrease in TNCs, but no other significant changes during the 5 years. In vitro measurements indicate that PBSCs can be successfully frozen and stored using a combination of DMSO and HES providing smaller amounts of DMSO and allowing simplified freezing and storage conditions.

  19. 21 CFR 146.148 - Reduced acid frozen concentrated orange juice.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Reduced acid frozen concentrated orange juice. 146... Canned Fruit Juices and Beverages § 146.148 Reduced acid frozen concentrated orange juice. (a) Reduced acid frozen concentrated orange juice is the food that complies with the requirements for composition...

  20. 75 FR 57501 - Frozen Warmwater Shrimp From Brazil, China, India, Thailand, and Vietnam

    Science.gov (United States)

    2010-09-21

    ... COMMISSION Frozen Warmwater Shrimp From Brazil, China, India, Thailand, and Vietnam AGENCY: United States... orders on frozen warmwater shrimp from Brazil, China, India, Thailand, and Vietnam. SUMMARY: The... orders on frozen warmwater shrimp from Brazil, China, India, Thailand, and Vietnam would be likely to...

  1. 75 FR 48724 - Frozen Warmwater Shrimp From Brazil, China, India, Thailand, and Vietnam

    Science.gov (United States)

    2010-08-11

    ... COMMISSION Frozen Warmwater Shrimp From Brazil, China, India, Thailand, and Vietnam AGENCY: United States... orders on frozen warmwater shrimp from Brazil, China, India, Thailand, and Vietnam. SUMMARY: The... orders on frozen warmwater shrimp from Brazil, China, India, Thailand, and Vietnam would be likely to...

  2. 7 CFR 319.56-12 - Importation of frozen fruits and vegetables.

    Science.gov (United States)

    2010-01-01

    ... HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Fruits and Vegetables § 319.56-12 Importation of frozen fruits and vegetables. Frozen fruits and vegetables may be imported... 7 Agriculture 5 2010-01-01 2010-01-01 false Importation of frozen fruits and vegetables. 319.56-12...

  3. 7 CFR 318.13-13 - Movement of frozen fruits and vegetables.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Movement of frozen fruits and vegetables. 318.13-13... Regulated Articles From Hawaii and the Territories § 318.13-13 Movement of frozen fruits and vegetables. Frozen fruits and vegetables may be certified for movement from Hawaii, Puerto Rico, Guam, the...

  4. 78 FR 2946 - United States Standards for Grades of Frozen Vegetables

    Science.gov (United States)

    2013-01-15

    ... Agricultural Marketing Service United States Standards for Grades of Frozen Vegetables AGENCY: Agricultural... Grades of Frozen Vegetables. AMS is proposing to replace the dual grade nomenclature with single letter... with turnips, and frozen mixed vegetables. DATES: Comments must be submitted on or before March 18...

  5. 76 FR 64001 - United States Standards for Grades of Frozen Okra

    Science.gov (United States)

    2011-10-17

    ... Service 7 CFR Part 52 United States Standards for Grades of Frozen Okra AGENCY: Agricultural Marketing... of Agriculture (USDA) has revised the United States Standards for Grades of Frozen Okra. The grade standards for frozen okra have been changed from a ``variables score point'' system to an ``individual...

  6. 7 CFR 58.318 - Butter, frozen or plastic cream melting machines.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Butter, frozen or plastic cream melting machines. 58... Service 1 Equipment and Utensils § 58.318 Butter, frozen or plastic cream melting machines. Shavers, shredders or melting machines used for rapid melting of butter, frozen or plastic cream shall be of...

  7. Expression pattern of vascular endothelial growth factor in canine folliculogenesis and its effect on the growth and development of follicles after ovarian organ culture.

    Science.gov (United States)

    Abdel-Ghani, M A; Shimizu, T; Suzuki, H

    2014-10-01

    In this study, the expressions of VEGF in dog follicles were detected by immunohistochemistry and the effects of VEGF treatment on the primordial to primary follicle transition and on subsequent follicle progression were examined using a dog ovary organ culture system. The frozen-thawed canine ovarian follicles within slices of ovarian cortical tissue were cultured for 7 and 14 days in presence or absence of VEGF. After culture, the ovaries were fixed, sectioned, stained and counted for morphologic analysis. The results showed that VEGF was expressed in the theca cells of antral follicles and in the granulosa cells nearest the oocyte in preantral follicle but not in granulosa cells of primordial and primary follicles; however, the VEGF protein was expressed in CL. After in vitro culture, VEGF caused a decrease in the number of primordial follicles and concomitant increase in the number of primary follicles that showed growth initiation and reached the secondary and preantral stages of development after 7 and 14 days. Follicular viability was also improved in the presence of VEGF after 7 and 14 days in culture. In conclusion, treatment with VEGF was found to promote the activation of primordial follicle development that could provide an alternative approach to stimulate early follicle development in dogs. © 2014 Blackwell Verlag GmbH.

  8. [Influence of high molecular weight polyethylene on viability of osteoblasts and new bone formation].

    Science.gov (United States)

    Ren, Gaohong; Lin, Angru; Pei, Guoxian; Hu, Basheng

    2006-02-01

    To investigate the influence of high molecular weight polyethylene (HMWP) on the viability of osteoblasts and new bone formation in the process of fracture healing, the osteoblasts derived from adult human bone marrow were cultured in HMWP maceration extract and normal culture medium. The viability of the osteoblasts was measured by MTT assay, and the function of the osteoblasts was detected by use of alkaline phosphatase test kit. The locked double-plating (steel plate and HMWP plate) was implanted and fixed at the artificial fracture of distal femur of dogs. Specimens were gained at 3, 6, 9 and 12 weeks postoperatively, examined with macroscopy, microscope and scanning electron microscope (SEM). The results showed that HMWP did no harm to osteoblasts. There is no significant difference in activities of proliferation and alkaline phosphatase between HMWP maceration extract and normal culture medium at each observation time of at 2,4,8, and 14 dyas (P>0. 05). Bone tissue under the implanted HMWP plate manifested no absorption; the new bones formed under the HMWP plate and gradually matured as time went on. It is demonstrated in this study that HMWP has no adverse influence on the viability of osteoblasts and new bone formation and it can be used as internal fixation implant in treating fractures.

  9. A key inactivation factor of HeLa cell viability by a plasma flow

    Energy Technology Data Exchange (ETDEWEB)

    Sato, Takehiko; Yokoyama, Mayo [Institute of Fluid Science, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai 980-8577 (Japan); Johkura, Kohei, E-mail: sato@ifs.tohoku.ac.jp [Department of Histology and Embryology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621 (Japan)

    2011-09-21

    Recently, a plasma flow has been applied to medical treatment using effects of various kinds of stimuli such as chemical species, charged particles, heat, light, shock wave and electric fields. Among them, the chemical species are known to cause an inactivation of cell viability. However, the mechanisms and key factors of this event are not yet clear. In this study, we focused on the effect of H{sub 2}O{sub 2} in plasma-treated culture medium because it is generated in the culture medium and it is also chemically stable compared with free radicals generated by the plasma flow. To elucidate the significance of H{sub 2}O{sub 2}, we assessed the differences in the effects of plasma-treated medium and H{sub 2}O{sub 2}-added medium against inactivation of HeLa cell viability. These two media showed comparable effects on HeLa cells in terms of the survival ratios, morphological features of damage processes, permeations of H{sub 2}O{sub 2} into the cells, response to H{sub 2}O{sub 2} decomposition by catalase and comprehensive gene expression. The results supported that among chemical species generated in a plasma-treated culture medium, H{sub 2}O{sub 2} is one of the main factors responsible for inactivation of HeLa cell viability. (fast track communication)

  10. Optimizing Photo-Encapsulation Viability of Heart Valve Cell Types in 3D Printable Composite Hydrogels.

    Science.gov (United States)

    Kang, Laura Hockaday; Armstrong, Patrick A; Lee, Lauren Julia; Duan, Bin; Kang, Kevin Heeyong; Butcher, Jonathan Talbot

    2017-02-01

    Photocrosslinking hydrogel technologies are attractive for the biofabrication of cardiovascular soft tissues, but 3D printing success is dependent on multiple variables. In this study we systematically test variables associated with photocrosslinking hydrogels (photoinitiator type, photoinitiator concentration, and light intensity) for their effects on encapsulated cells in an extrusion 3D printable mixture of methacrylated gelatin/poly-ethylene glycol diacrylate/alginate (MEGEL/PEGDA3350/alginate). The fabrication conditions that produced desired hydrogel mechanical properties were compared against those that optimize aortic valve or mesenchymal stem cell viability. In the 3D hydrogel culture environment and fabrication setting studied, Irgacure can increase hydrogel stiffness with a lower proportional decrease in encapsulated cell viability compared to VA086. Human adipose derived mesenchymal stem cells (HADMSC) survived increasing photoinitiator concentrations in photo-encapsulation conditions better than aortic valve interstitial cells (HAVIC) and aortic valve sinus smooth muscle cells (HASSMC). Within the range of photo-encapsulation fabrication conditions tested with MEGEL/PEGDA/alginate (0.25-1.0% w/v VA086, 0.025-0.1% w/v Irgacure 2959, and 365 nm light intensity 2-136 mW/cm2), the highest viabilities achieved were 95, 93, and 93% live for HASSMC, HAVIC, and HADMSC respectively. These results identify parameter combinations that optimize cell viability during 3D printing for multiple cell types. These results also indicate that general oxidative stress is higher in photocrosslinking conditions that induce lower cell viability. However, suppressing this increase in intracellular oxidative stress did not improve cell viability, which suggests that other stress mechanisms also contribute.

  11. Assessment of probiotic viability during Cheddar cheese manufacture and ripening using propidium monoazide-PCR quantification

    Directory of Open Access Journals (Sweden)

    Emilie eDesfossés-Foucault

    2012-10-01

    Full Text Available The use of a suitable food carrier such as cheese could significantly enhance probiotic viability during storage. The main goal of this study was to assess viability of commercial probiotic strains during Cheddar cheesemaking and ripening (four to six months by comparing the efficiency of microbiological and molecular approaches. Molecular methods such as quantitative PCR (qPCR allow bacterial quantification, and DNA-blocking molecules such as propidium monoazide (PMA select only the living cells’ DNA. Cheese samples were manufactured with a lactococci starter and with one of three probiotic strains (Bifidobacterium animalis subsp. lactis BB-12, Lactobacillus rhamnosus RO011 or Lactobacillus helveticus RO052 or a mixed culture containing B. animalis subsp. lactis BB-12 and L. helveticus RO052 (MC1, both lactobacilli strains (MC2 or all three strains (MC3. DNA extractions were then carried out on PMA-treated and non-treated cell pellets in order to assess PMA treatment efficiency, followed by quantification using the 16S rRNA gene, the elongation factor Tu gene (tuf or the transaldolase gene (tal. Results with intact/dead ratios of bacteria showed that PMA-treated cheese samples had a significantly lower bacterial count than non-treated DNA samples (P<0.005, confirming that PMA did eliminate dead bacteria from PCR quantification. For both quantification methods, the addition of probiotic strains seemed to accelerate the loss of lactococci viability in comparison to control cheese samples, especially when L. helveticus RO052 was added. Viability of all three probiotic strains was also significantly reduced in mixed culture cheese samples (P<0.0001, B. animalis subsp. lactis BB-12 being the most sensitive to the presence of other strains. However, all probiotic strains did retain their viability (log nine cfu/g of cheese throughout ripening. This study was successful in monitoring living probiotic species in Cheddar cheese samples through PMA-qPCR.

  12. Exercise regulates breast cancer cell viability

    DEFF Research Database (Denmark)

    Dethlefsen, Christine; Lillelund, Christian; Midtgaard, Julie

    2016-01-01

    Purpose: Exercise decreases breast cancer risk and disease recurrence, but the underlying mechanisms are unknown. Training adaptations in systemic factors have been suggested as mediating causes. We aimed to examine if systemic adaptations to training over time, or acute exercise responses......, in breast cancer survivors could regulate breast cancer cell viability in vitro. Methods: Blood samples were collected from breast cancer survivors, partaking in either a 6-month training intervention or across a 2 h acute exercise session. Changes in training parameters and systemic factors were evaluated...... and pre/post exercise-conditioned sera from both studies were used to stimulate breast cancer cell lines (MCF-7, MDA-MB-231) in vitro. Results: Six months of training increased VO2peak (16.4 %, p

  13. Candida sergipensis, a new asexual yeast species isolated from frozen pulps of tropical fruits.

    Science.gov (United States)

    Trindade, Rita C; Resende, Maria A; Pimenta, Raphael S; Lachance, Marc-André; Rosa, Carlos A

    2004-07-01

    Sixteen strains of the new yeast species Candida sergipensis have been isolated from frozen pulps of the tropical fruits umbú ( Spondias tuberosa Avr. Cam.) and mangaba ( Hancornia speciosa Gom.). Candida sergipensis was one of the prevalent species in the yeast community of these substrates. The new asexual ascomycetous yeast is phylogenetically related to Candida spandovensis and Candida sorbophila, species belonging to the Wickerhamiella clade, as evidenced by the sequences of the D1/D2 domains of their large subunit ribosomal DNAs. The species C. sergipensis and C. spandovensis can be separated on the basis of growth on 50% glucose agar, xylose and succinate, negative for the first species and positive for the second. The type culture is strain UFMG-R188 (CBS 9567).

  14. Artificial evolution by viability rather than competition.

    Directory of Open Access Journals (Sweden)

    Andrea Maesani

    Full Text Available Evolutionary algorithms are widespread heuristic methods inspired by natural evolution to solve difficult problems for which analytical approaches are not suitable. In many domains experimenters are not only interested in discovering optimal solutions, but also in finding the largest number of different solutions satisfying minimal requirements. However, the formulation of an effective performance measure describing these requirements, also known as fitness function, represents a major challenge. The difficulty of combining and weighting multiple problem objectives and constraints of possibly varying nature and scale into a single fitness function often leads to unsatisfactory solutions. Furthermore, selective reproduction of the fittest solutions, which is inspired by competition-based selection in nature, leads to loss of diversity within the evolving population and premature convergence of the algorithm, hindering the discovery of many different solutions. Here we present an alternative abstraction of artificial evolution, which does not require the formulation of a composite fitness function. Inspired from viability theory in dynamical systems, natural evolution and ethology, the proposed method puts emphasis on the elimination of individuals that do not meet a set of changing criteria, which are defined on the problem objectives and constraints. Experimental results show that the proposed method maintains higher diversity in the evolving population and generates more unique solutions when compared to classical competition-based evolutionary algorithms. Our findings suggest that incorporating viability principles into evolutionary algorithms can significantly improve the applicability and effectiveness of evolutionary methods to numerous complex problems of science and engineering, ranging from protein structure prediction to aircraft wing design.

  15. Fertilisation rate obtained with frozen-thawed boar semen supplemented with rosmarinic acid using a single insemination timed according to vulvar skin temperature changes.

    Science.gov (United States)

    Luño, Victoria; Gil, Lydia; Olaciregui, Maite; Grandía, Juan; Ansó, Trinidad; De Blas, Ignacio

    2015-03-01

    Artificial insemination (AI) of sows with frozen-thawed semen usually results in lower pregnancy rates and litter sizes than the use of liquid preserved semen. The present study evaluated the effectiveness of vulvar skin temperature changes as a predictor of ovulation in sows and determined the fertility rates obtained after AI with frozen-thawed semen supplemented with rosmarinic acid (RA). Semen was collected from mature boars and cryopreserved in experimental extenders supplemented with or without 105 μM of RA. Multiparous sows were inseminated with a single dose of semen when vulvar skin temperature decreased to a value below 35 °C. Intrauterine insemination was performed using 1.5 × 109 spermatozoa. The sows were slaughtered 48 h after AI and the embryos and oocytes were recovered from the oviducts. Total and progressive motility, viability and acrosome integrity were significantly (P < 0.05) higher in RA-supplemented semen samples compared with the control. Fertilisation occurred in all sows inseminated in the study, although there were no significant differences between the experimental groups. Sows inseminated with RA-supplemented semen showed a slight increase in the number of embryos recovered as compared to sows inseminated with control semen. In conclusion, insemination according to vulvar skin temperature changes resulted in successful fertilisation in all sows, although supplementation of the freezing media with RA did not improve the fertilising ability of frozen-thawed boar sperm.

  16. The relationship between sperm viability and DNA fragmentation rates.

    Science.gov (United States)

    Samplaski, Mary K; Dimitromanolakis, Apostolos; Lo, Kirk C; Grober, Ethan D; Mullen, Brendan; Garbens, Alaina; Jarvi, Keith A

    2015-05-14

    In humans, sperm DNA fragmentation rates have been correlated with sperm viability rates. Reduced sperm viability is associated with high sperm DNA fragmentation, while conversely high sperm viability is associated with low rates of sperm DNA fragmentation. Both elevated DNA fragmentation rates and poor viability are correlated with impaired male fertility, with a DNA fragmentation rate of >30% indicating subfertility. We postulated that in some men, the sperm viability assay could predict the sperm DNA fragmentation rates. This in turn could reduce the need for sperm DNA fragmentation assay testing, simplifying the infertility investigation and saving money for infertile couples. All men having semen analyses with both viability and DNA fragmentation testing were identified via a prospectively collected database. Viability was measured by eosin-nigrosin assay. DNA fragmentation was measured using the sperm chromosome structure assay. The relationship between DNA fragmentation and viability was assessed using Pearson's correlation coefficient. From 2008-2013, 3049 semen analyses had both viability and DNA fragmentation testing. A strong inverse relationship was seen between sperm viability and DNA fragmentation rates, with r=-0.83. If viability was ≤50% (n=301) then DNA fragmentation was ≥ 30% for 95% of the samples. If viability was ≥75% (n=1736), then the DNA fragmentation was ≤30% for 95% of the patients. Sperm viability correlates strongly with DNA fragmentation rates. In men with high levels of sperm viability≥75%, or low levels of sperm viability≤ 30%, DFI testing may be not be routinely necessary. Given that DNA fragmentation testing is substantially more expensive than vitality testing, this may represent a valuable cost-saving measure for couples undergoing a fertility evaluation.

  17. Princesa congelada? Uma leitura feminista de Frozen - uma aventura congelante

    Directory of Open Access Journals (Sweden)

    Jacqueline Sobral

    2016-02-01

    Full Text Available O presente artigo se vale das teorias da representação social para debater a construção simbólica do papel da mulher na sociedade, tendo como suporte as narrativas de contos de fada, em especial as histórias produzidas pela Disney. Partindo da hipótese de que o filme Frozen - uma aventura congelante (Frozen, 2014 representa uma leitura moderna dos tradicionais contos de fadas, propomos pensar o protagonismo da “princesa Elsa” como uma possível adoção de diretrizes feministas na construção de histórias infantis por um dos maiores conglomerados de mídia do mundo.

  18. The subzero microbiome: microbial activity in frozen and thawing soils.

    Science.gov (United States)

    Nikrad, Mrinalini P; Kerkhof, Lee J; Häggblom, Max M

    2016-06-01

    Most of the Earth's biosphere is characterized by low temperatures (soils are not yet well understood, although the threat of microbial contribution to positive feedback of carbon flux is substantial. To date, several studies have examined microbial community dynamics in frozen soils and permafrost due to changing environmental conditions, and some have undertaken the complicated task of characterizing microbial functional groups and how their activity changes with changing conditions, either in situ or by isolating and characterizing macromolecules. With increasing temperature and wetter conditions microbial activity of key microbes and subsequent efflux of greenhouse gases also increase. In this review, we aim to provide an overview of microbial activity in seasonally frozen soils and permafrost. With a more detailed understanding of the microbiological activities in these vulnerable soil ecosystems, we can begin to predict and model future expectations for carbon release and climate change. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  19. Intraoperative visualization, frozen section, and permanent pathology in endometrial cancer

    Directory of Open Access Journals (Sweden)

    Soheila Aminimoghaddam

    2014-12-01

    Conclusion: Although the sample size of the studied population was small but our study results support the previous data and suggest that in early stages and low grade tumors, gross examination and frozen section diagnosis are conveniently predictive of lymph node metastasis. These data might be useful for prediction of tumor invasion using frozen section and gross examination in low grade tumors and early stages and for doing complete surgical staging and lymph node sampling. However the im-portance of surgical staging always must be considered in patients who need systemat-ic lymphadenectomy. In overall these data might help to come up with new guidelines for surgical risk assessment in endometrial cancer.

  20. Relationships among frozen-thawed semen fertility, physical parameters, certain routine sperm characteristics and testosterone in breeding Murrah buffalo (Bubalus bubalis bulls

    Directory of Open Access Journals (Sweden)

    A. K. Singh

    2014-09-01

    Full Text Available Aim: The present study was carried out to examine the relationships among frozen-thawed semen fertility, physical parameters, seminal quality, and testosterone concentration in Murrah buffalo bulls. Materials and Methods: A total of 30 breeding Murrah buffalo bulls (either progeny tested or under progeny testing program were randomly selected from two government bull farms in Punjab. None of the bulls selected for this study had any preceding physical abnormality. A field fertility trial was conducted to determine the first service conception rate (FSCR. The number of females inseminated per bull semen was 10. All the bulls were inspected for structural soundness, measurement of scrotal circumference, testicular biometry, and internal pelvic area (IPA. Frozen-thawed semen was evaluated for total motility, progressive motility, viability, concentration, abnormality, and hypo-osmotic swelling test (HOST. Testosterone was estimated in blood plasma, seminal plasma as well as frozen-thawed semen extracts for establishing relationship. Results: The FSCR was 48% in the bulls having a scrotal circumference of ≥44 cm, although, there was no significant correlation between FSCR and scrotal circumference. Similarly, no consistent relationship existed between sperm concentration and scrotal circumference. A positive correlation was observed between IPA and FSCR (r=0.294. Of the six post-thaw seminal components (total motility, progressive motility, viability, HOST (%, total abnormality and concentration only total motility had a high significant (p<0.01 correlation with FSCR (r=0.694. Varied correlations existed between other seminal parameters and fertility. Using a simple regression analysis, the post-thaw motility, IPA, prepuce length and testosterone (independent variables combined to explain approximately 62% of the variation in the FSCR (dependent variable. Conclusion: The present study indicated that despite low to high correlations between seminal

  1. Grid-group cultural theory: an introduction

    NARCIS (Netherlands)

    Mamadouh, V.

    1999-01-01

    This article offers an introduction to grid-group cultural theory (also known as grid-group analysis, Cultural Theory or theory of socio-cultural viability), an approach that has been developed over the past thirty years in the work of the British anthropologists Mary Douglas and Michael Thompson,

  2. Enhanced Viability of Endothelial Colony Forming Cells in Fibrin Microbeads for Sensor Vascularization

    Directory of Open Access Journals (Sweden)

    Jarel K. Gandhi

    2015-09-01

    Full Text Available Enhanced vascularization at sensor interfaces can improve long-term function. Fibrin, a natural polymer, has shown promise as a biomaterial for sensor coating due to its ability to sustain endothelial cell growth and promote local vascularization. However, the culture of cells, particularly endothelial cells (EC, within 3D scaffolds for more than a few days is challenging due to rapid loss of EC viability. In this manuscript, a robust method for developing fibrin microbead scaffolds for long-term culture of encapsulated ECs is described. Fibrin microbeads are formed using sodium alginate as a structural template. The size, swelling and structural properties of the microbeads were varied with needle gauge and composition and concentration of the pre-gel solution. Endothelial colony-forming cells (ECFCs were suspended in the fibrin beads and cultured within a perfusion bioreactor system. The perfusion bioreactor enhanced ECFCs viability and genome stability in fibrin beads relative to static culture. Perfusion bioreactors enable 3D culture of ECs within fibrin beads for potential application as a sensor coating.

  3. Viewing the Disney Movie Frozen through a Psychodynamic Lens.

    Science.gov (United States)

    Kowalski, Christopher; Bhalla, Ruchi

    2015-10-14

    The Disney movie Frozen is the fifth highest grossing movie of all time. In order to better understand this phenomenon and to hypothesize as to why the movie resonated so strongly with audiences, we have interpreted the movie using psychodynamic theory. We pay particular attention to the themes of puberty, adolescence and sibling relationships and discuss examples of ego defenses that are employed by the lead character in relation to these concepts.

  4. Measuring Negative Pore Pressures in Partially Frozen Saturated Soils

    OpenAIRE

    Holten, Johannes Gaspar

    2017-01-01

    Freezing of soil is an issue which has many implications on modern infrastructure, in which frost heave plays a pivotal role. During freezing the behavior of the soil and the flow of water is altered. In an engineering perspective, it is important to grasp the driving forces behind these behavioral changes. The main contributor to frost heave is the development of a large negative pore pressure in the unfrozen water in partially frozen fine-grained soil, termed cryosuction. The suction leads...

  5. Novel Membrane System to Automatically Deglycerolize Thawed Frozen Human Blood

    Science.gov (United States)

    1997-04-01

    equivalent to the current Haemonetics 115 device. • Washing frozen/thawed RBCs with the WUP I and WUP II/B/ H devices yielded glycerol washouts of...RHeaton, A, Mesbah -Karimi N, Bross J. Vox Sang 12. Blood Bank Week 1995;12(23):3 36 List of Tables Table 1: Sequence of Events for TBPS Console Table...2..... *1. . ...i.... .,..6. . . ............ 6 ..... 4 E...,..6.... H ..o

  6. Angular correlation of annihilation photons in frozen aqueous solutions

    DEFF Research Database (Denmark)

    Milosevic-Kvajic, M.; Mogensen, O. E.; Kvajic, G.

    1972-01-01

    Linear‐slit angular correlation curves were obtained at about −140°C for frozen aqueous solutions of HF, HCl, HBr, HI, NH3, FeCl2, FeCl3, NaI, H2SO4, NHO3, MnSO4, KMnO4, K2Cr2O7, NaOH, and LiOH. We found no appreciable influence of a 4% concentration of the last seven impurities. Only halide...

  7. Surface-Atmosphere Moisture Coupling in Eurasian Frozen Ground Regions

    Science.gov (United States)

    Frauenfeld, O. W.; Ford, T.

    2014-12-01

    Permafrost represents an impermeable barrier to moisture, resulting in a saturated or near-saturated surface layer during the warm season in many continuous and discontinuous permafrost zones. These surface conditions could lead to enhanced convection and precipitation during the warm season, and significant local recycling of moisture. In areas underlain by sporadic or isolated permafrost, or in seasonally frozen areas, the moisture can drain away more readily, resulting in much drier soil conditions. As climate change causes frozen ground degradation, this will thus also alter the patterns of atmospheric convection, moisture recycling, and the hydrologic cycle in high-latitude land areas. In this study, we analyze evaporative fraction (EF) as a proxy for evapotranspiration, and precipitation from the Modern-Era Retrospective analysis for Research and Applications (MERRA-land) reanalysis dataset. We focus on 1979-2012 and document patterns and changes in EF over the Eurasian high latitudes. We find strong, positive April EF trends over the study period, particularly in the Lena River Basin, 80% of which is underlain by continuous permafrost. In fact, these significant positive trends in spring EF are strongest over continuous permafrost across the Eurasian high latitudes, but negative for sporadic and isolated permafrost. In addition, we find a strong, statistically significant relationship between EF anomalies and the probability of subsequent precipitation over the Lena Basin during April. This association therefore suggests a potential land-atmosphere coupling between frozen ground and precipitation. As the permafrost and seasonally frozen ground distribution changes in the future, this will likely have repercussions for the Arctic hydrologic cycle.

  8. Cu(II) chloride complexes in frozen ethanol solution

    Energy Technology Data Exchange (ETDEWEB)

    Plyusnin, V.F.; Usov, O.M.

    1985-12-01

    This paper examines the photochemical properties of Cu(II) chloride complexes in frozen organic matrices. It was necessary to determine the spectroscopic parameters and conditions of the existence of individual complexes at 77 K. Ethanol was saturated with gaseous HC1, the solubility of which is appreciably greater than that of such salts as LiC1, NaC1, or KC1. The spectroscopic parameters of the complexes are determined and discussed.

  9. Characterization of a frozen shoulder model using immobilization in rats.

    Science.gov (United States)

    Kim, Du Hwan; Lee, Kil-Ho; Lho, Yun-Mee; Ha, Eunyoung; Hwang, Ilseon; Song, Kwang-Soon; Cho, Chul-Hyun

    2016-12-08

    The objective of this study was to investigate serial changes for histology of joint capsule and range of motion of the glenohumeral joint after immobilization in rats. We hypothesized that a rat shoulder contracture model using immobilization would be capable of producing effects on the glenohumeral joint similar to those seen in patients with frozen shoulder. Sixty-four Sprague-Dawley rats were randomly divided into one control group (n = 8) and seven immobilization groups (n = 8 per group) that were immobilized with molding plaster for 3 days, or for 1, 2, 3, 4, 5, or 6 weeks. At each time point, eight rats were euthanized for histologic evaluation of the axillary recess and for measurement of the abduction angle. Infiltration of inflammatory cells was found in the synovial tissue until 2 weeks after immobilization. However, inflammatory cells were diminished and fibrosis was dominantly observed in the synovium and subsynovial tissue 3 weeks after immobilization. From 1 week after immobilization, the abduction angle of all immobilization groups at each time point was significantly lower than that of the control group. Our study demonstrated that a rat frozen shoulder model using immobilization generates the pathophysiologic process of inflammation leading to fibrosis on the glenohumeral joint similar to that seen in patients with frozen shoulder. This model was attained within 3 weeks after immobilization. It may serve as a useful tool to investigate pathogenesis at the molecular level and identify potential target genes that are involved in the development of frozen shoulder.

  10. On the compressibility and temperature boundary of warm frozen soils

    Science.gov (United States)

    Qi, Jilin; Dang, Boxiang; Guo, Xueluan; Sun, Xiaoyu; Yan, Xu

    2017-04-01

    A silty-clay obtained along the Qinghai-Tibetan railway and a standard Chinese sand were taken as study objects. Saturated frozen soil samples were prepared for testing. Step-load was used and confined compression was carried out on the soils under different temperatures. Compression index and pseudo-preconsolidation pressure (PPC) were obtained. Unlike unfrozen soils, PPC is not associated with stress history. However, it is still the boundary of elastic and plastic deformations. Different compression indexes can be obtained from an individual compression curve under pressures before and after PPC. The parameters at different thermal and stress conditions were analyzed. It is found that temperature plays a critical role in mechanical behaviours of frozen soils. Efforts were then made on the silty-clay in order to suggest a convincing temperature boundary in defining warm frozen soil. Three groups of ice-rich samples with different ice contents were prepared and tested under confined compression. The samples were compressed under a constant load and with 5 stepped temperatures. Strain rates at different temperatures were examined. It was found that the strain rate at around -0.6°C increased abruptly. Analysis of compression index was performed on the data both from our own testing program and from the literature, which showed that at about -1°C was a turning point in the curves for compression index against temperature. Based on both our work and taking into account the unfrozen water content vs. temperature, the range of -1°C to -0.5°C seems to be the temperature where the mechanical properties change greatly. For convenience, -1.0°C can be defined as the boundary for warm frozen soils.

  11. THE EFFECTS OF AMBIENT DISSOCIATION ON FROZEN HYPERSONIC STAGNATION FLOW.

    Science.gov (United States)

    The effects of ambient dissociation on frozen flow in the vicinity of a stagnation point are discussed within the framework of the thin shock-layer...recombination heat transfer. The general effects of ambient dissociation at low Reynolds numbers are discussed, and special consideration is given to the...effects of ambient dissociation on a variety of flow quantities are calculated. The discussion includes an estimate of the extent of gas-phase reaction at low Reynolds numbers. (Author)

  12. Stability of Frozen Rat Plasma Containing Different Antibiotics

    Science.gov (United States)

    Berti, Maria A.; Maccari, Mario

    1975-01-01

    The antibiotic activity was determined at different intervals of time on plasma samples, taken from rats treated with a certain number of commonly used antibiotics, and kept at −20 C up to 8 weeks. The results of the microbiological assays demonstrate that the stability of the antibiotics in the frozen plasma decreases in the following order: oxytetracycline > cephalexin, streptomycin, erythromycin > demeclocycline > ampicillin, amoxycillin > penicillin G, cephaloridine, rolitetracycline, and tetracycline. PMID:1211917

  13. Bacterial content in the intestine of frozen common carp Cyprinus ...

    African Journals Online (AJOL)

    The quantitative and qualitative analyses of bacterial flora associated with the intestine of common carp Cyprinus carpio in fresh and frozen storage conditions for 16 months was determined. Aerobic plate counts (APC) ranged from 1.8 ± 2.7 × 109 to 1.1 ± 2.9 x 104 colony forming unit (CFU) g-1, 2.0 ± 2.1 x 1010 to 6.1 ± 1.4 ...

  14. Metyrapone, an inhibitor of cytochrome oxidases, does not affect viability in a neuroblastoma cell model of bilirubin toxicity

    Directory of Open Access Journals (Sweden)

    Maria N. Naguib Leerberg

    2014-01-01

    Conclusion: Herein we have shown that metyrapone does not increase cell death in neuroblastoma cells in culture exposed to bilirubin. Our results question the relationship between the oxidative mechanism evaluated by spectrophotometry and cell viability. Our findings add to the discussion on whether bilirubin oxidation represents a potentially important protective mechanism in neurons challenged by hyperbilirubinemia.

  15. Viability, Apoptosis, Proliferation, Activation, and Cytokine Secretion of Human Keratoconus Keratocytes after Cross-Linking

    Directory of Open Access Journals (Sweden)

    Xuefei Song

    2015-01-01

    Full Text Available Purpose. The purpose of this study was to determine the impact of cross-linking (CXL on viability, apoptosis, proliferation, activation, and cytokine secretion of human keratoconus (KC keratocytes, in vitro. Methods. Primary KC keratocytes were cultured in DMEM/Ham’s F12 medium supplemented with 10% FCS and underwent UVA illumination (370 nm, 2 J/cm2 during exposure to 0.1% riboflavin and 20% Dextran in PBS. Twenty-four hours after CXL, viability was assessed using Alamar blue assay; apoptosis using APO-DIRECT Kit; proliferation using ELISA-BrdU kit; and CD34 and alpha-smooth muscle actin (α-SMA expression using flow cytometry. Five and 24 hours after CXL, FGFb, HGF, TGFβ1, VEGF, KGF, IL-1β, IL-6, and IL-8 secretion was measured using enzyme-linked-immunoabsorbent assay (ELISA. Results. Following CXL, cell viability and proliferation decreased (P0.06. Five hours after CXL, FGFb secretion increased significantly (P=0.037; however no other cytokine secretion differed significantly from controls after 5 or 24 hours (P>0.12. Conclusions. Cross-linking decreases viability, triggers apoptosis, and inhibits proliferation, without an impact on multipotent hematopoietic stem cell transformation and myofibroblastic transformation of KC keratocytes. CXL triggers FGFb secretion of KC keratocytes transiently (5 hours, normalizing after 24 hours.

  16. In Vitro Pollen Viability and Pollen Germination in Cherry Laurel (Prunus laurocerasus L.

    Directory of Open Access Journals (Sweden)

    Melekber Sulusoglu

    2014-01-01

    Full Text Available Pollen quality is important for growers and breeders. This study was carried out to determine in vitro pollen viability and pollen germination in seven genotypes of cherry laurel (Prunus laurocerasus L.. Two pollen viability tests, TTC (2,3,5-triphenyl tetrazolium chloride and IKI (iodine potassium iodide, were used. Pollen traits of genotypes were studied using an in vitro medium containing 0%, 5%, 10%, 15%, and 20% sucrose to determine the best sucrose concentrations for germination. In the second step, the germinated pollen was counted 1, 4, 6, 10, 12, 24, and 48 hours later until there was no further germination. The viability rates were different according to genotypes and tests used. The IKI and TTC staining tests and pollen germination had low correlation (r2 = 0.0614 and r2 = 0.0015, resp.. Painted pollen rate was higher and pollen was well-stained with IKI test and pollen viability estimated with TTC staining test was better than that estimated with the IKI staining test. 15% sucrose gave the best germination rates in most of the genotypes. Pollen germination rates were recorded periodically from one hour to 48 hours in 15% sucrose and the results showed that pollen germination rates increased after 6 hours of being placed in culture media.

  17. Viability, apoptosis, proliferation, activation, and cytokine secretion of human keratoconus keratocytes after cross-linking.

    Science.gov (United States)

    Song, Xuefei; Stachon, Tanja; Wang, Jiong; Langenbucher, Achim; Seitz, Berthold; Szentmáry, Nóra

    2015-01-01

    The purpose of this study was to determine the impact of cross-linking (CXL) on viability, apoptosis, proliferation, activation, and cytokine secretion of human keratoconus (KC) keratocytes, in vitro. Primary KC keratocytes were cultured in DMEM/Ham's F12 medium supplemented with 10% FCS and underwent UVA illumination (370 nm, 2 J/cm(2)) during exposure to 0.1% riboflavin and 20% Dextran in PBS. Twenty-four hours after CXL, viability was assessed using Alamar blue assay; apoptosis using APO-DIRECT Kit; proliferation using ELISA-BrdU kit; and CD34 and alpha-smooth muscle actin (α-SMA) expression using flow cytometry. Five and 24 hours after CXL, FGFb, HGF, TGFβ1, VEGF, KGF, IL-1β, IL-6, and IL-8 secretion was measured using enzyme-linked-immunoabsorbent assay (ELISA). Following CXL, cell viability and proliferation decreased (P 0.06). Five hours after CXL, FGFb secretion increased significantly (P = 0.037); however no other cytokine secretion differed significantly from controls after 5 or 24 hours (P > 0.12). Cross-linking decreases viability, triggers apoptosis, and inhibits proliferation, without an impact on multipotent hematopoietic stem cell transformation and myofibroblastic transformation of KC keratocytes. CXL triggers FGFb secretion of KC keratocytes transiently (5 hours), normalizing after 24 hours.

  18. The effect of Aloe vera gel on viability of dental pulp stem cells.

    Science.gov (United States)

    Sholehvar, Fatemeh; Mehrabani, Davood; Yaghmaei, Parichehr; Vahdati, Akbar

    2016-10-01

    Dental pulp stem cells (DPSCs) can play a prominent role in tissue regeneration. Aloe vera L. (Liliaceae) contains the polysaccharide of acemannan that was shown to be a trigger factor for cell proliferation, differentiation, mineralization, and dentin formation. This study sought to determine the viability of DPSCs in Aloe vera in comparison with Hank's balanced salt solution (HBSS). Twelve rabbits underwent anesthesia, and their incisor teeth were extracted; the pulp tissue was removed, chopped, treated with collagenase and plated in culture flasks. DPSCs from passage 3 were cultured in 24-well plates, and after 3 days, the culture media changed to 10, 25, 50, and 100% concentrations of Aloe vera at intervals of 45 and 90 min and 3 and 6 h. Distilled water was used as negative and HBSS as positive control for comparison. The cell morphology, viability, population doubling time (PDT), and growth kinetics were evaluated. RT-PCR was carried out for characterization and karyotyping for chromosomal stability. Aloe vera showed a significant higher viability than HBSS (74.74%). The 50% Aloe vera showed higher viability (97.73%) than other concentrations. PDT in 50% concentration was 35.1 h and for HBSS was 49.5 h. DPSCs were spindle shaped and were positive for CD73 and negative for CD34 and CD45. Karyotyping was normal. Aloe vera as an inexpensive and available herb can improve survival of avulsed or broken teeth in emergency cases as a transfer media. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  19. Effect of alcohol on insulin secretion and viability of human pancreatic islets

    Directory of Open Access Journals (Sweden)

    Nikolić Dragan

    2017-01-01

    Full Text Available Introduction/Objective. There are controversial data in the literature on the topic of effects of alcohol on insulin secretion, apoptosis, and necrosis of the endocrine and exocrine pancreas. The goal of this research was to determine how alcohol affects the insulin secretion and viability of human adult pancreatic islets in vitro during a seven-day incubation. Methods. Human pancreatic tissue was digested with Collagenase XI, using a non-automated method. Cultures were incubated in Roswell Park Memorial Institute (RPMI medium containing alcohol (10 μl of alcohol in 100 ml of medium. Insulin stimulation index (SI and viability of the islets were determined on the first, third, and seventh day of cultivation. Results. Analysis of the viability of the islets showed that there wasn’t significant difference between the control and the test group. In the test group, viability of the cultures declined with the time of incubation. SI of the test group was higher compared to the control group, by 50% and 25% on the first and third day of cultivation, respectively. On the seventh day, insulin secretion was reduced by 25%. The difference was not statistically significant (p > 0.05. In the test group, significant decline in insulin secretion was found on the third and seventh day of incubation (p ≤ 0.05. Conclusion. Alcohol can increase or decrease insulin secretion of islets cultures, which may result in an inadequate response of pancreatic β-cells to blood glucose, leading to insulin resistance, and increased risk of developing type 2 diabetes. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 41002

  20. Bacteriological Studies on Meat Pies and Frozen Prepared Dinners

    Science.gov (United States)

    Alexander, D. C.; Tittiger, F.

    1971-01-01

    A bacteriological survey of meat pies, frozen prepared dinners and entrees indicated that their bacterial populations were related to the components, the environment and handling in manufacture. Results were correlated with microbiological examinations of ingredients, line samples and plant environmental samples. Total bacterial counts, coliform counts and Staphylococcus counts of the finished product were low. Coliform organisms found were primarily Aerobacter spp. originating with the frozen vegetables. Enterococci were also found in the frozen vegetables and finished product. Both enterococci and Aerobacter organisms were recovered from the plant environment where they appeared to multiply. Many of the plant personnel carried coagulase-positive staphylococci in the throat or nasal cavity, and this was indicated as a possible source for these organisms found in low numbers in the finished product. Cooking of the products produced sterility except for the survival of some spore forming Bacillus spp. The presence of a poultry slaughtering operation adjacent to the manufacturing area was considered to contribute to microbial contamination of the air in the plant. PMID:4251416

  1. Surgical Tips in Frozen Abdomen Management: Application of Coliseum Technique

    Directory of Open Access Journals (Sweden)

    Ioannis D. Kyriazanos

    2015-01-01

    Full Text Available Wound dehiscence is a serious postoperative complication, with an incidence of 0.5–3% after primary closure of a laparotomy incision, and represents an acute mechanical failure of wound healing. Relatively recently the concept of “intentional open abdomen” was described and both clinical entities share common pathophysiological and clinical pathways (“postoperative open abdominal wall”. Although early reconstruction is the target, a significant proportion of patients will develop adhesions between abdominal viscera and the anterolateral abdominal wall, a condition widely recognized as “frozen abdomen,” where delayed wound closure appears as the only realistic alternative. We report our experience with a patient who presented with frozen abdomen after wound dehiscence due to surgical site infection and application of the “Coliseum technique” for its definitive surgical management. This novel technique represents an innovative alternative to abdominal exploration, for cases of “malignant” frozen abdomen due to peritoneal carcinomatosis. Lifting the edges of the surgical wound upwards and suspending them under traction by threads from a retractor positioned above the abdomen facilitates approach to the peritoneal cavity, optimizes exposure of intra-abdominal organs, and prevents operative injury to the innervation and blood supply of abdominal wall musculature, a crucial step for subsequent hernia repair.

  2. PRESERVING QUALITY OF FROZEN GREEN PEAS DURING LONG TIME STORAGE

    Directory of Open Access Journals (Sweden)

    FELICIA DIMA

    2012-06-01

    Full Text Available The purpose of this study was to analyze the degree which fresh green peas maintain their technological properties and nutritional components during long-term storage, for a period of 24 months, in a defined condition of the air, at maximum -18°C. Peas were blanched, at four different durations of this operation and the same temperature of the blanching water, 95°C. The peas were frozen after end stored. In conditions of long-term storage of frozen peas, it has been found a considerable loss of the initial content of vitamin C and damage of chlorophyll (total, a and b, under the action of chlorophyllase, causing some gray compounds. The conditions of the storage in frozen stage produced a distortion of color and sensorial qualities, higher as the storage temperature was higher than prescribed. It was found that the best results in preserving quality of peas during long-term storage (24 months were obtained for the blanched samples at 4 minutes at 95°C. The reducing of the content of vitamin C, chlorophyll a and b, and sensorial qualities was the lowest, overall.

  3. DNA integrity of fresh and frozen canine epididymal spermatozoa.

    Science.gov (United States)

    Varesi, Sara; Vernocchi, Valentina; Morselli, Maria Giorgia; Luvoni, Gaia Cecilia

    2014-12-01

    The aims of this study were to evaluate: (1) the effect of cryopreservation on DNA fragmentation of canine epididymal spermatozoa, and (2) the potential protective effect of melatonin on post-thaw sperm quality (motility, morphology, acrosomal and DNA integrity). Epididymal spermatozoa were collected after orchiectomy of ten dogs. Sperm samples were frozen in the presence or absence of melatonin (1mM). DNA fragmentation index (percentage of spermatozoa with fragmented DNA) was similar in fresh samples (3.3±3.6) and samples frozen with (4.2±3.8) or without (3.6±3.7) melatonin. Sperm motility was significantly (psperm motility. Proportions of spermatozoa with normal morphology were similar in fresh and frozen samples, irrespective of the presence of melatonin in the extender. Acrosome integrity was significantly decreased (pepididymal spermatozoa was not affected by the freezing procedure, and the presence of melatonin did not preserve motility and acrosome integrity which were adversely affected by cryopreservation. The evaluation of DNA status of thawed gametes is particularly relevant for epididymal spermatozoa since these spermatozoa are usually stored and used in assisted reproductive techniques. Copyright © 2014 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  4. Sperm viability staining in ecology and evolution: potential pitfalls

    DEFF Research Database (Denmark)

    Holman, Luke

    2009-01-01

    a number of interesting results, it has some potential pitfalls that have rarely been discussed. In the present paper, I review the major findings of ecology and evolution studies employing sperm viability staining and outline the method's principle limitations. The key problem is that the viability assay......The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced...

  5. Assessing the Viability of Tiger Subpopulations in a Fragmented Landscape

    National Research Council Canada - National Science Library

    Matthew Linkie; Guillaume Chapron; Deborah J. Martyr; Jeremy Holden; Nigel Leader-Williams

    2006-01-01

    .... This study aimed to provide such information for tigers in the Kerinci Seblat (KS) region, Sumatra, by identifying and assessing subpopulation viability under different management strategies. 2...

  6. Fault Detection and Isolation using Viability Theory and Interval Observers

    Science.gov (United States)

    Ghaniee Zarch, Majid; Puig, Vicenç; Poshtan, Javad

    2017-01-01

    This paper proposes the use of interval observers and viability theory in fault detection and isolation (FDI). Viability theory develops mathematical and algorithmic methods for investigating the adaptation to viability constraints of evolutions governed by complex systems under uncertainty. These methods can be used for checking the consistency between observed and predicted behavior by using simple sets that approximate the exact set of possible behavior (in the parameter or state space). In this paper, fault detection is based on checking for an inconsistency between the measured and predicted behaviors using viability theory concepts and sets. Finally, an example is provided in order to show the usefulness of the proposed approach.

  7. The effects of storage conditions on the viability of ...

    African Journals Online (AJOL)

    The effects of storage conditions on the viability of enteropathogenics bacteria in biobanking of human stools: Cases of Yersinia enterocolitica, Salmonella enterica Typhimurium and Vibrio cholerae O: 1.

  8. FEM-based oxygen consumption and cell viability models for avascular pancreatic islets

    Directory of Open Access Journals (Sweden)

    Buchwald Peter

    2009-04-01

    Full Text Available Abstract Background The function and viability of cultured, transplanted, or encapsulated pancreatic islets is often limited by hypoxia because these islets have lost their vasculature during the isolation process and have to rely on gradient-driven passive diffusion, which cannot provide adequate oxygen transport. Pancreatic islets (islets of Langerhans are particularly susceptible due to their relatively large size, large metabolic demand, and increased sensitivity to hypoxia. Here, finite element method (FEM based multiphysics models are explored to describe oxygen transport and cell viability in avascular islets both in static and in moving culture media. Methods Two- and three-dimensional models were built in COMSOL Multiphysics using the convection and diffusion as well as the incompressible Navier-Stokes fluid dynamics application modes. Oxygen consumption was assumed to follow Michaelis-Menten-type kinetics and to cease when local concentrations fell below a critical threshold; in a dynamic model, it was also allowed to increase with increasing glucose concentration. Results Partial differential equation (PDE based exploratory cellular-level oxygen consumption and cell viability models incorporating physiologically realistic assumptions have been implemented for fully scaled cell culture geometries with 100, 150, and 200 μm diameter islets as representative. Calculated oxygen concentrations and intra-islet regions likely to suffer from hypoxia-related necrosis obtained for traditional flask-type cultures, oxygen-permeable silicone-rubber membrane bottom cultures, and perifusion chambers with flowing media and varying incoming glucose levels are presented in detail illustrated with corresponding colour-coded figures and animations. Conclusion Results of the computational models are, as a first estimate, in good quantitative agreement with existing experimental evidence, and they confirm that during culture, hypoxia is often a problem for

  9. Storage Viability and Optimization Web Service

    Energy Technology Data Exchange (ETDEWEB)

    Stadler, Michael; Marnay, Christ; Lai, Judy; Siddiqui, Afzal; Limpaitoon, Tanachai; Phan, Trucy; Megel, Olivier; Chang, Jessica; DeForest, Nicholas

    2010-10-11

    Non-residential sectors offer many promising applications for electrical storage (batteries) and photovoltaics (PVs). However, choosing and operating storage under complex tariff structures poses a daunting technical and economic problem that may discourage potential customers and result in lost carbon and economic savings. Equipment vendors are unlikely to provide adequate environmental analysis or unbiased economic results to potential clients, and are even less likely to completely describe the robustness of choices in the face of changing fuel prices and tariffs. Given these considerations, researchers at Lawrence Berkeley National Laboratory (LBNL) have designed the Storage Viability and Optimization Web Service (SVOW): a tool that helps building owners, operators and managers to decide if storage technologies and PVs merit deeper analysis. SVOW is an open access, web-based energy storage and PV analysis calculator, accessible by secure remote login. Upon first login, the user sees an overview of the parameters: load profile, tariff, technologies, and solar radiation location. Each parameter has a pull-down list of possible predefined inputs and users may upload their own as necessary. Since the non-residential sectors encompass a broad range of facilities with fundamentally different characteristics, the tool starts by asking the users to select a load profile from a limited cohort group of example facilities. The example facilities are categorized according to their North American Industry Classification System (NAICS) code. After the load profile selection, users select a predefined tariff or use the widget to create their own. The technologies and solar radiation menus operate in a similar fashion. After these four parameters have been inputted, the users have to select an optimization setting as well as an optimization objective. The analytic engine of SVOW is LBNL?s Distributed Energy Resources Customer Adoption Model (DER-CAM), which is a mixed

  10. THE 'FROZEN' HEART OF THE CONTINENT: PLACE-BRANDING WINNIPEG IN CANADIAN CRIME FILMS

    Directory of Open Access Journals (Sweden)

    James Gacek

    2016-08-01

    Full Text Available Brands play an imperative role when representing places, as culture and the people who live and create it are at the core of the brand. Aside from promoting experiences and destinations, place brands enhance culture and identity, and orchestrate space and atmosphere to create a particular ambiance in which societies are immersed. Over the past century, the representation of Canada in fictional film has generally emphasized its northerly location, its imagined wilderness landscape, and inevitably, its harsh winter climate. For films that focus on crime, justice is frequently imagined along the lines of the cinematic western, with frontier lawmen bringing order and the reach of the state to remote frontier outposts. For much of the twentieth century, this cultural depiction of Winnipeg justice remained frozen and unchallenged. However, beginning in the latter decades of the twentieth century, a growing number of independent Canadian crime films have taken up these familiar Hollywood tropes in ways that reflect, refract, and sometimes challenge the dominant construction of crime and justice. I examine how these films construct crime and the places that foster crime by emphasizing aspects of racialized identity, class, and the branding dynamics of the city of Winnipeg. Far from the rugged northwest of mid-century Hollywood cinema, crime is relocated to the contemporary city, and while still shaped by the endless Canadian winter, justice is frequently fraught, and the power of law and order distant and unable to extend the civilizing effects of the state to these new urban frontiers.

  11. Assessment of a simple, non-toxic Alamar blue cell survival assay to monitor tomato cell viability.

    Science.gov (United States)

    Byth, H A; Mchunu, B I; Dubery, I A; Bornman, L

    2001-01-01

    The Alamar Blue (AB) assay, which incorporates a medox indicator that changes colour or fluorescence in response to metabolic activity, is commonly used to assess quantitatively the viability and/or proliferation of mammalian cells and micro-organisms. In this study the AB assay was adapted for the determination of the viability of plant cells. Cell suspension cultures of tomato, Lycopersicon esculentum, L., with differing viabilities, served as the experimental model for a comparison of the AB assay with the conventional 2,3,5-triphenyltetrazolium chloride (TTC) viability assay. The AB assay showed a sigmoidal relationship between cell viability and AB reduction (as quantified by spectrofluorometry or spectrophotometry), which was similar to that obtained using the TTC assay. Both assays detected a significant reduction in cell viability after 48 h exposure to virulent Ralstonia solanacearum (biovar III), while the TTC assay, in addition, revealed cell proliferation in control cells from 24 to 72 h. The TTC assay detected cell proliferation over a wider range of cell densities, while the AB assay was more rapid and versatile whilst being non-toxic and thus allowing subsequent cell analysis.

  12. Assessment of sperm nuclear quality after in vitro maturation of fresh or frozen/thawed mouse pre-pubertal testes.

    Science.gov (United States)

    Oblette, A; Rives, N; Dumont, L; Rives, A; Verhaeghe, F; Jumeau, F; Rondanino, C

    2017-10-01

    Is nuclear quality of in vitro generated spermatozoa from fresh or frozen/thawed pre-pubertal mouse testes similar to that of their in vivo counterparts? The production of spermatozoa with aneuploidy, DNA fragmentation or chromatin condensation defects was not significantly increased in organotypic cultures compared to in vivo controls. Although murine spermatozoa have been produced in vitro from pre-pubertal testes, their nuclear DNA integrity has never been investigated. Fresh and frozen/thawed testicular fragments from 6 to 7 days postpartum (dpp) mice were cultured for 30 days. Testicular tissues were frozen by controlled slow freezing (CSF) or solid surface vitrification (SSV). In total, 30 fresh, 30 CSF, 30 SSV testes were used for in vitro maturation and 6 testes from 36 to 37 dpp mice were used as in vivo controls. Murine spermatozoa were extracted from pooled in vitro cultured testicular fragments and from in vivo controls. Sperm aneuploidy was analyzed by fluorescence in situ hybridization (FISH), DNA fragmentation by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling, chromatin condensation by aniline blue staining, telomere length and number by quantitative FISH, DNA oxidation by immunocytochemical detection of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Because of the low spermatogenic yield in cultures, a hundred spermatozoa extracted from pooled tissues were examined and compared to their in vivo counterparts. Most of spermatozoa generated in vitro and in vivo were haploid, contained unfragmented DNA and normally condensed chromatin. A similar proportion of spermatozoa with aneuploidy, DNA fragmentation or chromatin condensation defects was found in cultures and in vivo. No significant difference in telomere length was found within the nuclei of in vitro and in vivo generated spermatozoa. However, the number of telomere spots was lower in gametes obtained from cultures of fresh, CSF and SSV testes than in their natural counterparts (P

  13. Microscale Electro-Hydrodynamic Cell Printing with High Viability.

    Science.gov (United States)

    He, Jiankang; Zhao, Xiang; Chang, Jinke; Li, Dichen

    2017-12-01

    Cell printing has gained extensive attentions for the controlled fabrication of living cellular constructs in vitro. Various cell printing techniques are now being explored and developed for improved cell viability and printing resolution. Here an electro-hydrodynamic cell printing strategy is developed with microscale resolution (95%). Unlike the existing electro-hydrodynamic cell jetting or printing explorations, insulating substrate is used to replace conventional semiconductive substrate as the collecting surface which significantly reduces the electrical current in the electro-hydrodynamic printing process from milliamperes (>0.5 mA) to microamperes (printed cells. The smallest width of the electro-hydrodynamically printed hydrogel filament is 82.4 ± 14.3 µm by optimizing process parameters. Multiple hydrogels or multilayer cell-laden constructs can be flexibly printed under cell-friendly conditions. The printed cells in multilayer hydrogels kept alive and gradually spread during 7-days culture in vitro. This exploration offers a novel and promising cell printing strategy which might benefit future biomedical innovations such as microscale tissue engineering, organ-on-a-chip systems, and nanomedicine. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Effect of different concentrations of soybean lecithin and virgin coconut oil in Tris-based extender on the quality of chilled and frozen-thawed bull semen

    Directory of Open Access Journals (Sweden)

    A. A. Tarig

    2017-06-01

    Full Text Available Aim: The objective of this study was to evaluate the effects of different concentrations of soybean lecithin (SL and virgin coconut oil (VCO in Tris-based extender on chilled and frozen-thawed bull semen quality parameters. Materials and Methods: A total of 24 ejaculates were collected from four bulls via an electroejaculator. Semen samples were diluted with 2% VCO in Tris-based extender which consists of various concentrations of SL (1, 1.25, 1.5, and 1.75%. A 20% egg yolk in Tris used as a positive control (C+. The diluted semen samples were divided into two fractions; one for chilling which were stored at 4°C for 24, 72, and 144 h before evaluated for semen quality parameters. The second fraction used for freezing was chilled for 3 h at 4°C, packed into 0.25 mL straws and then cryopreserved in liquid nitrogen. The samples were then evaluated after 7 and 14 days. Chilled and frozen semen samples were thawed at 37°C and assessed for general motility using computer-assisted semen analysis, viability, acrosome integrity and morphology (eosin-nigrosin stain, membrane integrity, and lipid peroxidation using thiobarbituric acid reaction test. Results: The results showed that all the quality parameters assessed were significantly (p<0.05 improved at 1.5% SL concentration in chilled semen. Treatment groups of 1, 1.25, 1.5, and 1.75% SL were higher in quality parameters than the control group (C+ in chilled semen. However, all the quality parameters in frozen-thawed semen were significantly higher in the C+ than the treated groups. Conclusion: In conclusion, supplementation of 1.5% SL in 2% VCO Tris-based extender enhanced the chilled bull semen. However, there was no marked improvement in the frozen-thawed quality parameters after treatment.

  15. Detection and Viability of Lactococcus lactis throughout Cheese Ripening

    Science.gov (United States)

    Cocolin, Luca

    2014-01-01

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

  16. Incorporating evolutionary processes into population viability models.

    Science.gov (United States)

    Pierson, Jennifer C; Beissinger, Steven R; Bragg, Jason G; Coates, David J; Oostermeijer, J Gerard B; Sunnucks, Paul; Schumaker, Nathan H; Trotter, Meredith V; Young, Andrew G

    2015-06-01

    We examined how ecological and evolutionary (eco-evo) processes in population dynamics could be better integrated into population viability analysis (PVA). Complementary advances in computation and population genomics can be combined into an eco-evo PVA to offer powerful new approaches to understand the influence of evolutionary processes on population persistence. We developed the mechanistic basis of an eco-evo PVA using individual-based models with individual-level genotype tracking and dynamic genotype-phenotype mapping to model emergent population-level effects, such as local adaptation and genetic rescue. We then outline how genomics can allow or improve parameter estimation for PVA models by providing genotypic information at large numbers of loci for neutral and functional genome regions. As climate change and other threatening processes increase in rate and scale, eco-evo PVAs will become essential research tools to evaluate the effects of adaptive potential, evolutionary rescue, and locally adapted traits on persistence. © 2014 Society for Conservation Biology.

  17. The effects of inclusion of exogenous phospholipid in Tris diluent with different level of egg yolk on the viability of bull spermatozoa

    Directory of Open Access Journals (Sweden)

    P Situmorang

    2002-10-01

    Full Text Available This study has been conducted to evaluate the effect of inclusion of phospholipid in Tris-diluent containing different level of egg yolk on the viability of bull spermatozoa after chilling and freezing. Semen was collected by means of artificial vagina, diluted in Tris-diluent to get a final concentration 100 x 106 spermatozoa/ml. Diluted semen was cooled to 50C for 45-60 minute, equiliberated in those temperature for 4 hours and frozen by placing the straw 5 cm above surface of liquid nitrogen for 10 minutes. The experiments was factorial designed with two level of phospholipid (0 and 0.5 mm and 4 concentration of egg yolk (0, 5, 10 and 20% v/v. The viability of spermatozoa was assessed by evaluating the percentage of motile, live and condition of apical ridge after the temperature reduced to 50C, stored at those temperature for 3 and 7 days and after thawing. Inclusion of phospholipid and level of egg yolk in Tris diluent significantly increased the viability of spermatozoa for both chilling and deepfreezing. The mean percentage of motile, live and intact apical ridge for 3 and 7 days of storage times at 50C, were significantly higher (P<0.05 in diluent containing phospholipid than without phospholipid. After thawing, the mean percentage motile, live and intact apical ridges were significantly higher (P<0.05 in a diluent containing phospholipid (49.9; 60.2 and 60.0 than those without phospholipid (39.1; 54.1 and 51.5. The effects of interaction between phospholipid and level of egg yolk on the viability of spermatozoa was not significant for both chilling and freezing. Level of egg yolk significantly (P<0.05 affected the viability of spermatozoa where the optimal level of egg yolk was 10% v/v for chilled semen and the higher level (20% v/v was needed for frozen semen. In conclusion, Tris diluent containing 0,5 mM phosphatidyl coline with 10 or 20% egg yolk gave a best protection for chilled and frozen semen respectively.

  18. THE PROPORTION OF X AND Y SPERM, VIABILITY AND MOTILITY OF RAM SPERMATOZOA AFTER SEPARATED WITH WHITE EGG ALBUMIN

    Directory of Open Access Journals (Sweden)

    Moh Takdir

    2017-02-01

    The aim of this research was to determine the proportion, viability and motility of X and Y ram spermatozoa separated with egg white albumin. Sperm samples derived from Garut ram, which was collected by using an artificial vagina. Observations were made on spermatozoa fraction above and below each medium fraction treatment. There are treatment egg white albumin as separation medium, each medium consisting of fractions top and bottom fraction with different concentration: 1 P0 = sperma before separation (control; 2 P1 = 10% above fraction + 30% lower fraction; P2 = 25% + 45%; P3 = 25% + 75%. Data proportion of X and Y, viability and motility were analyzed statistically by Completely Randomized Design patern in the direction followed by Duncan’s Multiple Range Test for data with a real difference. Separation with egg white albumin affect significantly increased the proportion of spermatozoa X and Y (P≤0.05, but tends to decrease the viability and motility of spermatozoa.The proportion of spermatozoa X and Y was highest in treatment P3,76.76% of spermatozoa X (fraction above 25% and 79.81% spermatozoa Y (75% lower fraction, with an average viability obtained respectively 68,9% (fraction above and 59,7% (bottom fraction, motility 77,5% (fraction above dan 84,0% (bottom fraction. It was concluded that the egg white albumin is very effective in changing the proportions of X and Y ram sperm with the quality of spermatozoa after separation feasible for applications insemination or processed into frozen semen.   (Keywords: Garut ram, White egg albumin, Spermatozoa X and Y

  19. Effect of storage duration, storage temperature, and diluent on the viability and fertility of fresh ram sperm.

    Science.gov (United States)

    O'Hara, L; Hanrahan, J P; Richardson, L; Donovan, A; Fair, S; Evans, A C O; Lonergan, P

    2010-03-01

    Cervical artificial insemination (AI) in sheep with fresh semen yields a much higher pregnancy rate than when frozen-thawed semen is used, and consequently frozen semen is only acceptable for laparoscopic insemination. The short life span of fresh semen is a major constraint on the use of AI in genetic improvement programs for sheep. The main objective of this study was to examine the effects of storage conditions on viability and fertilization ability of fresh ram (Ovis aries) semen up to 72h postcollection. Experiment 1 was designed to evaluate the effect of diluent type (standard skim milk, AndroMed, OviPro, and INRA 96) and storage temperature (5 degrees C and 15 degrees C) on the motility and viability of fresh ram semen. Storage temperature, irrespective of diluent, had a significant effect on both motility and viability. Storage at 5 degrees C maintained acceptable motility and viability up to 72h compared with that of storage at 15 degrees C. In Experiment 2, the penetrating ability of fresh ram semen, diluted in either skim milk, AndroMed, or INRA 96, was assessed using artificial mucus. Flat capillary tubes containing artificial mucus were suspended in 250muL semen at a sperm concentration of 20x10(6)/mL. Semen was stored at 5 degrees C and tested after 6, 24, 48, and 72h. There was a significant diluent by time interaction. In Experiment 3, the fertilizing ability of fresh ram semen stored at 5 degrees C was evaluated in vitro. Fresh semen (diluted in either skim milk, AndroMed, or INRA 96) was added to matured ewe oocytes at 6, 24, or 72h after semen collection. Cleavage rate was recorded at 48h postinsemination, and blastocyst development was recorded on Days 6 to 9. There was a significant treatment effect on cleavage and blastocyst rates; insemination of semen stored for 24h resulted in higher rates than those for storage at 72h. In Experiment 4, the fertilizing ability of fresh ram semen was evaluated in vivo. Semen was diluted in INRA 96, stored at

  20. The effect of alginate and hyaluronate on the viability and function of immunoisolated neonatal rat islets.

    Science.gov (United States)

    Velten, F; Laue, C; Schrezenmeir, J

    1999-11-01

    Recent observations suggest that the extracellular matrix plays a role as regards the viability and morphological integrity in long-term culture of Langerhans islets. For the present study we encapsulated neonatal rat islets without extracellular matrix (WEM), with alginate solution (AL) and with hyaluronate solution (HY) into cuprophane hollow fibers. Function was tested after week 1 and 5. The insulin release of WEM encapsulated islets decreased significantly during the culture period. In contrast to this, AL and HY embedded islets had stable secretion values throughout the whole cultivation. Histological examination confirmed that viability of HY and of AL embedded islets differed significantly from that of WEM encapsulated islets. Furthermore, HY seems to be a more advantageous environment to immunoisolated islets than AL. Both the insulin secretion values and the viability of HY embedded islets were higher than of AL embedded islets. We conclude that an extracellular matrix is important for immunoisolated islets, to maintain their function and morphological integrity and that HY is especially suitable for this application.

  1. Evaluation of tissue reaction, cell viability and cytokine production induced by Sealapex Plus

    Directory of Open Access Journals (Sweden)

    João Eduardo Gomes-Filho

    2011-08-01

    Full Text Available OBJECTIVE: The aim of this study was to investigate the effects of mineral trioxide aggregate (MTA, Sealapex, and a combination of Sealapex and MTA (Sealapex Plus on the reaction of subcutaneous connective tissue of rats, and on cell viability and cytokine production in mouse fibroblasts. MATERIAL AND METHODS: The tissue reaction was carried out with dentin tubes containing the materials implanted in the dorsal connective tissue of rats. The histological analysis was performed after 7 and 30 days. Millipore culture plate inserts with polyethylene tubes filled with materials were placed into 24-well cell culture plates with mouse fibroblasts to evaluate the cell viability by MTT assay. ELISA assays were also performed after 24 h of exposure of the mouse fibroblasts to set material disks. RESULTS: Histopathologic examination showed Von Kossa-positive granules that were birefringent to polarized light for all the studied materials at the tube openings. No material inhibited the cell viability in the in vitro test. It was detected IL-6 production in all root-end filling materials. MTA and Sealapex Plus induced a slight raise of mean levels of IL-1β. CONCLUSIONS: The results suggest that Sealapex Plus is biocompatible and stimulates the mineralization of the tissue.

  2. 40 CFR 405.80 - Applicability; description of the ice cream, frozen desserts, novelties and other dairy desserts...

    Science.gov (United States)

    2010-07-01

    ... PRODUCTS PROCESSING POINT SOURCE CATEGORY Ice Cream, Frozen Desserts, Novelties and Other Dairy Desserts Subcategory § 405.80 Applicability; description of the ice cream, frozen desserts, novelties and other dairy... manufacture of ice cream, ice milk, sherbert, water ices, stick confections, frozen novelties products, frozen...

  3. Viability and molecular authentication of Coccidioides immitis strains from culture collection of the Instituto Oswaldo Cruz, Rio de Janeiro, Brazil Viabilidade e autenticação molecular de cepas de Coccidioides immitis da coleção de culturas do Instituto Oswaldo Cruz, Rio de Janeiro, Brasil

    Directory of Open Access Journals (Sweden)

    Claudia C.F. Bezerra

    2006-06-01

    Full Text Available Twenty Coccidioides immitis strains were evaluated. Only 5 of the 20 strains kept under mineral oil maintained their viability while all 5 subcultures preserved in water remained viable and none of the 13 subcultures kept in soil were viable. A 519 bp PCR product from the csa gene confirmed the identity of the strains.Vinte cepas de Coccidioides immitis foram avaliadas. Cinco das 20 cepas preservadas sob óleo mineral mantiveram-se viáveis, todas as 5 subculturas preservadas em água permaneceram viáveis e nenhuma das 13 subculturas mantidas em solo foi viável. Um produto de PCR de 519 pb do gene csa confirmou a identidade das cepas.

  4. Effect of freezing time on the quality of Indian mackerel (Rastrelliger kanagurta) during frozen storage.

    Science.gov (United States)

    Lakshmisha, I P; Ravishankar, C N; Ninan, G; Mohan, C O; Gopal, T K S

    2008-09-01

    The present study aims to find the effect of freezing methods on the quality of mackerel (Rastrelliger kanagurta) in commercial plate and air blast freezers during freezing and subsequent frozen storage (-18 degrees C). Total time for freezing was significantly different (P plate and air blast freezers (90 and 220 min, respectively). This difference in the freezing time could be attributed to the varied quality of the 2 samples. Upon freezing, the moisture content decreased in air blast frozen samples compared to plate freezer where protein content decreased in both the samples. Upon freezing and during frozen storage, lipid oxidation products (peroxide value, thiobarbutiric acid value, and free fatty acid value) and volatile bases (total volatile base nitrogen and trimethyl amine nitrogen) showed an increasing trend in both the samples with values slightly higher in air blast frozen samples compared to plate frozen samples. The total plate counts showed a significantly (P plate frozen samples. The taste and overall acceptability was significantly different (P plate frozen samples compared to air blast frozen samples on 3rd month. Both samples were in acceptable condition up to 3 mo but the plate frozen samples quality was slightly better than the air blast frozen samples.

  5. Zona pellucida damage to human embryos after cryopreservation and the consequences for their blastomere survival and in-vitro viability.

    Science.gov (United States)

    Van Den Abbeel, E; Van Steirteghem, A

    2000-02-01

    The study objective was to quantify zona pellucida (ZP) damage in cryopreserved human embryos. The influence of two different freezing containers was investigated, and the influence of freezing damage on the survival and viability of the embryos evaluated. ZP damage did not differ according to whether embryos originated from in-vitro fertilization (IVF) cycles or from IVF cycles in association with intracytoplasmic sperm injection (ICSI). The freezing container, however, significantly influenced the occurrence of ZP damage after cryopreservation. More damage was observed when the embryos were frozen-thawed using plastic cryovials than using plastic mini-straws (16.6% versus 2.3%; P plastic mini-straws. The further cleavage of frozen-thawed embryos suitable for transfer was not different whether there was ZP damage or not; however, it was higher when there was 100% blastomere survival as compared with when some blastomeres were damaged (79.0% versus 43.7%; P plastic mini-straws. In conclusion, the aim of a cryopreservation programme should be to have as many fully intact embryos as possible after thawing. Increased ZP damage might indicate a suboptimal cryopreservation procedure.

  6. [Study on the pollen viability and stigma receptivity of Chrysanthemum morifolium 'Fubaiju'].

    Science.gov (United States)

    Xu, Lei; Liu, Chang-Li; Wang, Hui-Di; Chen, Ke-Li

    2012-10-01

    To study the blooming habits, pollen viability and stigma receptivity of Chrysanthemum morifolium and provide theoretical basis for its breeding. Explored the blooming habits by dynamic observation on the process of blossom, evaluated the pollen viability by germination in vitro culture method and estimated stigma receptivity by benzidine-hydrogen peroxide method. About the pollen viability, there were no significant differences between the flowers which in the same round of the capitulum; Tubular flowers in the center of a capitulum were significantly higher than that on the edge; In the morning pollen vitality gradually raised, during 11: 00 - 14: 00 maintained the highest, and then gradually decreased; Tubular flower began to loose powder on the third day, during 4th - 6th day the pollen viability was highest, respectively was 35.12%, 39.89%, 38.12%, then gradually decreased, on the 15th day was only 7.41%, finally turned into wither. Regard to the stigma receptivity, the center of a capitulum were significantly higher than that on the edge, outer edge ligulate flower had no receptivity; Revealed the strongest during 13: 00 - 14:00 in one day; During the 5th - 7th day was the strongest after flowering. The regulation of the stigma secreted mucus existed great consistency with the stigma receptivity, namely the stigma receptivity usually was strong when it secreted large number mucus. Understand the blossom habits of Chrysanthemun morifolium, as well as the dynamic changes regulation of pollen viability and stigma receptivity during its blossom, which could be used to select the flowers in a capitulum which are on the more suitable period and position for artificial pollination and hybridization breeding research.

  7. Establishing guidelines to retain viability of probiotics during spray drying

    NARCIS (Netherlands)

    Perdana, J.A.; Fox, M.B.; Boom, R.M.; Schutyser, M.A.I.

    2014-01-01

    We present a model-based approach to map processing conditions suitable to spray dry probiotics with minimal viability loss. The approach combines the drying history and bacterial inactivation kinetics to predict the retention of viability after drying. The approach was used to systematically assess

  8. 37 CFR 1.807 - Viability of deposit.

    Science.gov (United States)

    2010-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2010-07-01 2010-07-01 false Viability of deposit. 1.807... COMMERCE GENERAL RULES OF PRACTICE IN PATENT CASES Biotechnology Invention Disclosures Deposit of Biological Material § 1.807 Viability of deposit. (a) A deposit of biological material that is capable of...

  9. Studies On Fermentation, Alcohol Production And Viability In ...

    African Journals Online (AJOL)

    The reverse was true in the sugarcane bagasse medium. Yeasts with high viability tended to have high alcohol production ability in the sucrose medium and vice-versa. KEY WORDS: Alcohol production; fermentation; induced mutants; Saccharomyces cerevisiae; viability. Global Journal of Pure and Applied Sciences ...

  10. The Economy and Democracy: Viability and Challenges for ...

    African Journals Online (AJOL)

    The Economy and Democracy: Viability and Challenges for Sustainable Democratisation in Nigeria. ... Economic and Policy Review ... the viability for developing sustainable democracy in Nigeria against the background of the country's enormous economic potentials and the economic reforms introduced following the ...

  11. Pollen viability and germination in Jatropha ribifolia and Jatropha ...

    African Journals Online (AJOL)

    The aim of this work is to assess pollen viability using the staining technique and in vitro germination with different concentrations of sucrose in Jatropha ribifolia and Jatropha mollissima, contributing to the knowledge of the reproductive biology and subsidizing their conservation, management and utilization. Pollen viability ...

  12. Viability, Advantages and Design Methodologies of M-Learning Delivery

    Science.gov (United States)

    Zabel, Todd W.

    2010-01-01

    The purpose of this study was to examine the viability and principle design methodologies of Mobile Learning models in developing regions. Demographic and market studies were utilized to determine the viability of M-Learning delivery as well as best uses for such technologies and methods given socioeconomic and political conditions within the…

  13. Recent developments in the use of viability dyes and quantitative PCR in the food microbiology field.

    Science.gov (United States)

    Elizaquível, P; Aznar, R; Sánchez, G

    2014-01-01

    The increase in foodborne outbreaks highlights the need for rapid, sensitive and specific methods for food safety monitoring, enabling specific detection and quantification of viable foodborne pathogens. Real-time PCR (qPCR) combined with the use of viability dyes, recently introduced, fulfils all these requirements. The strategy relies on the use of DNA-binding molecules such as propidium monoazide (PMA) or ethidium monoazide (EMA) as sample pretreatment previous to the qPCR. These molecules permeate only membrane-compromised cells and have successfully been applied for different types of foodborne pathogens, including bacteria and viruses. Moreover, those dyes have been explored to monitor different food manufacturing processes as an alternative to classical cultural methods. In this review, state-of-the-art information regarding viability PCR (v-PCR) is compiled. © 2013 The Society for Applied Microbiology.

  14. Verification of cell viability at progressively higher scanning forces using a hybrid atomic force and fluorescence microscope.

    Science.gov (United States)

    Barnes, C A; O'Hagan, B M G; Howard, C V; McKerr, G

    2007-11-01

    The prudent use of the atomic force microscope as a supra-vital live cell imaging tool requires that cell viability must be determined before and after scanning. Complementary optical techniques in conjunction with the fluorescent dyes rhodamine-123 and ethidium homodimer have been used within this study to determine cell viability after increasing loads are applied in contact mode. Guideline force ranges for five commonly cultured cell lines, human squamous carcinoma (A431), fibroblast, HeLa, Potorous tridactylis (PtK2) and rat intestinal epithelial (RIE) cells are given.

  15. [Hemodialysis improves the subendocardial viability ratio].

    Science.gov (United States)

    De Blasio, Antonella; Sirico, Maria; Di Micco, Lucia; Di Iorio, Biagio

    2013-01-01

    The subendocardial viability ratio (SEVR), a parameter introduced by Buckberg, represents a non-invasive measure of myocardial perfusion related to left ventricular work. AIM. The aim of this study was to verify if dialysis may determine modifications of SEVR and how these modifications are modulated in the 2-day interdialytic period. METHODS.We studied 54 subjects of mean age 6314 years and receiving dialysis for 3215 months. Exclusion criteria were diabetes, resistant hypertension and peripheral vascular diseases and intradialytic hypotension evidenced during the study dialysis session. Pulse wave velocity and SEVR assessments were performed during the third dialysis session of the week, before (pre-HD) and after (post-HD) dialysis, in 2-day interdialytic period after and at the beginning of the following dialysis session. RESULTS.Dialysis reduces PWV, in particular the tertile with the lowest PWV presents the highest percentage reduction (-26%) compared with the second and the third tertiles. In the same way, dialysis leads to an increase of SEVR and patients in the tertile with the highest SEVR values maintain high SEVR values during dialysis and in the interdialytic period. Patients with severe vascular calcifications present higher PWV value and lower SEVR value. CONCLUSIONS.The results of present study demonstrate that ultrafiltration improves PWV (with a mean reduction of 16%) and SEVR (increase of 13%) and that the severity of vascular calcifications influences the effect of ultrafiltration on these two parameters. More studies are certainly necessary to verify our findings. Considered the higher mortality of patients with higher SEVR, it would be important to understand if new dialytic strategies are needed in patients with higher PVW and lower SEVR values.

  16. Viability and functional integrity of washed platelets

    Energy Technology Data Exchange (ETDEWEB)

    Pineda, A.A.; Zylstra, V.W.; Clare, D.E.; Dewanjee, M.K.; Forstrom, L.A.

    1989-07-01

    The viability and functional integrity of saline- and ACD-saline-washed platelets were compared with those of unwashed platelets. After template bleeding time (TBT) was measured, 15 healthy volunteers underwent plateletpheresis and ingested 600 mg of aspirin. Autologous /sup 111/In-labeled platelets were transfused: unwashed (n = 5), washed with 0.9 percent saline solution (SS) (n = 5), and washed with a buffered 12.6 percent solution of ACD-A in 0.9 percent saline solution (n = 5). After transfusion, we measured TBT at 1, 4, and 24 hours; platelet survival at 10 minutes and 1, 4, and 24 hours and daily for 6 days; and the percentage of uptake in liver and spleen by quantitative whole-body radionuclide scintigraphy at 24 and 190 hours. We found that saline washing affected platelet recovery, 23.47 +/- 12 percent (p less than 0.001) as compared to 52.43 +/- 17 percent (p less than 0.002) for ACD-saline and 73.17 +/- 8 percent for control; that saline washing resulted in a greater liver uptake than control and ACD-saline-washed platelets (31.9 +/- 8% (p less than 0.001) vs 17.7 +/- 4.1 and 19.3 +/- 2.1% (p greater than 0.1), respectively); that, unlike control and ACD-saline-washed platelets, saline-washed platelets did not shorten bleeding time; and that neither type of washing affected survival. Although ACD-saline washing affects recovery, it also results in intact function, normal survival, higher recovery than SS platelets, and no significant liver uptake.

  17. Bacteriology of unshelled frozen blue swimming crab (Portunus pelagicus).

    Science.gov (United States)

    Giuffrida, Alessandro; Ziino, Graziella; La Paola, Roberto; Bottari, Teresa; Panebianco, Antonio

    2004-04-01

    In this study, 45 (10 whole specimens and 35 frozen claws) frozen samples of Portunus pelagicus imported into Sicily (Italy) from the west coast of Africa were examined to assess their bacteriological characteristics and suitability for consumption. Bacteriological examination was performed on two subsamples for each whole crab. The first was the body and claw muscle; the second was a pool of viscera and gills. In the case of frozen claws, each muscle claw was a sample. An aerobic plate count at 30 degrees C (mesophilic aerobic plate count [MAPC]) and 18 degrees C (psychrotrophic aerobic plate count [PAPC]) for 3 days, sulfite-reducing anaerobes, total coliforms, Escherichia coli, enterococci, and Aeromonas spp. were enumerated. Detection of halophilic Vibrio spp. was also performed using salt polymixin broth as an enrichment medium and thiosulfate citrate bile salts sucrose agar as a selective medium; a further morphological and biochemical identification of suspected colonies was performed. The bacterial load of muscle and viscera and gills was low. The MAPC ranged from 0.78 to 3.26 log CFU/g, and the PAPC ranged from 0.48 to 2.41 log CFU/g. Vibrio spp., Aeromonas spp., sulfite-reducing anaerobes, and E. coli were never isolated from muscles or viscera and gills. In contrast to the findings of others, this study showed good bacteriological quality of crabs imported into Sicily from the west coast of Africa. This study also demonstrated the positive influence of the characteristics of environment of origin and postharvest handling hygiene; these parameters could be useful in the context of the application of the hazard analysis critical control point system to this production.

  18. Number density of liquid inclusions formed in frozen aqueous electrolyte.

    Science.gov (United States)

    Hashimoto, Takuya; Harada, Makoto; Nojima, Shuichi; Okada, Tetsuo

    2013-10-07

    Frozen aqueous chlorides (≤50 mM) are characterized by using confocal fluorescence microscopy and small angel X-ray scattering (SAXS). The former method allows us to determine the size of a liquid inclusion formed in the ice matrix at temperatures above the eutectic point of the system (t(eu)). Isolated liquid inclusions of a uniform size are formed when the temperature of a frozen electrolyte increases past t(eu). The size of the liquid inclusions depends on the observation temperature as well as on the concentration (c(salt)) and type of salt dissolved in the original unfrozen solution. However, the number density of liquid inclusions is almost constant and independent of these experimental parameters, particularly when an electrolyte is frozen in liquid nitrogen. Salt accumulation can then occur at the imperfections of the ice crystals. The occurrence probability of the imperfections is independent of the nature of an incorporated salt. The amount of a salt confined in each inclusion ranges from 7 to 240 fmol, depending on c(salt). SAXS measurements provide information on the size of individual salt crystals formed at temperatures below t(eu). The radius of gyration of a salt crystal ranges from 2 to 2.8 nm, and does not depend significantly on c(salt). Thus, each inclusion is formed from 10(6)-10(9) nanocrystals, which can act as seeds. When doped ice is prepared at higher temperatures, for example -16 °C, the isolation of liquid inclusions is not sufficient and coalescence occurs more easily upon an increase in temperature or cs(alt). However, when c(salt) is lower than 10 mM, the number density of liquid inclusions is almost constant, irrespective of the freezing temperature. Copyright © 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Levels of Mercury in Persian Gulf Frozen Fish Species

    Directory of Open Access Journals (Sweden)

    Parisa Ziarati

    2017-03-01

    Full Text Available Severe discharge of sewage and industrial effluents into the Persian Gulf leads to the deposition of various types of heavy metals, especially lead and mercury, in the muscles of fish. Total mercury and methylmercury contents were determined in the edible parts (muscle tissue, fillet of two different most popular frozen fish species from the Persian Gulf to ascertain whether the concentrations exceeded the maximum level fixed by the European Commission or not. During the period from October 2015 to June 2016, a total of 150 frozen fish packaged samples were randomly collected from the recognized supermarkets in Tehran province, Iran. The mercury (Hg concentration of samples was determined by atomic absorption spectrophotometer using a mercuric hydride system (MHS 10 and also by direct mercury analyzer (DMA. High concentration of total Hg was found in a Carcharhinus dussumie brand (0.91 ± 0.12 μg/g while the lowest level was detected in Pomadasys furcatus (0.29 ± 0.02 μg/g. In current study the mean concentrations of Mercury in all studied frozen fish samples were 0.79 ± 0.11 µg/g that means Hg levels were above 0.5 μg/g, which is the maximum standard level recommended by Joint FAO/WHO/Expert Committee on Food Additives (JECFA. In 13% of Pomadasys and in 47.2 % of Carcharhinus fish samples total mercury concentrations exceeded the maximum level fixed by the European Commission. All samples had also mean Hg concentrations that exceeded EPA's established safety level of 0.3 μg/g.

  20. Seminal plasma proteins interacting with sperm surface revert capacitation indicators in frozen-thawed ram sperm.

    Science.gov (United States)

    Ledesma, Alba; Fernández-Alegre, Estela; Cano, Adriana; Hozbor, Federico; Martínez-Pastor, Felipe; Cesari, Andreína

    2016-10-01

    This study was conducted to evaluate the effects of interacting seminal plasma proteins (iSPP) obtained by AV or EE on frozen-thawed ram sperm in order to test the hypothesis whether this fraction could be sufficient to emulate the effect of complete seminal plasma (SP). Additionally, we evaluated whether these proteins have a differential effect between spermatozoa from high and low fertility rams and between breeding and non-breeding seasons. We assessed sperm motility, quality parameters (intracellular reactive oxygen species, membrane fluidity, plasma membrane permeability and mitochondrial activity) and capacitation status. The main findings from this work were: i) iSPP had no effect on sperm motility, whereas SP (AV or EE) addition produced the highest values of total motility (74.13±2.99 and 72.27±2.99 for AV and EE, respectively) and progressive motility (64.97±2.64 and 63.73±2.64 for AV and EE, respectively); ii) iSPP had no effect on sperm quality parameters (p>0.05), but whole SP improved all parameters evaluated. Moreover, SP collected by AV yielded significantly higher viability (44.60±2.87) and sperm with stable plasma membrane (44.56±2.49) comparing with the addition of SP collected by EE (35.80±2.47 and 36.67±1.71, respectively); iii) iSPP and SP collected by EE, but not by AV, reverted molecular signals of capacitation as protein tyrosine phosphorylation caused by freezing temperatures; iv) there were no effects of fertility or season in sperm quality parameters evaluated. This study demonstrated that, although the iSPP have a clear decapacitating effect, including the ability to revert cryo-capacitation indicators, they are not sufficient to emulate the effects of complete SP regarding sperm functional parameters. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Frozen and broadband slow light in coupled periodic nanowire waveguides.

    Science.gov (United States)

    Gutman, Nadav; Dupree, W Hugo; Sun, Yue; Sukhorukov, Andrey A; de Sterke, C Martijn

    2012-02-13

    We develop novel designs enabling slow-light propagation with vanishing group-velocity dispersion ("frozen light") and slow-light with large delay-bandwidth product, in periodic nanowires. Our design is based on symmetry-breaking of periodic nanowire waveguides and we demonstrate its vailidy through two- and three-dimensional simulations. The slow-light is associated with a stationary inflection point which appears through coupling between forward and backward waveguide modes. The mode coupling also leads to evanescent modes, which enable efficient light coupling to the slow mode.

  2. Exploring the outcomes in studies of primary frozen shoulder

    DEFF Research Database (Denmark)

    Rodgers, Sara; Brealey, Stephen; Jefferson, Laura

    2014-01-01

    considered important; and re-examined papers previously obtained for a systematic review of patients' views of interventions for frozen shoulder to investigate their views on outcomes. RESULTS: Thirty-one studies investigated the outcomes range of movement (28 studies), pain (22), function and disability (22......), adverse events (13), quality of life (7) and other outcomes (5). Many different types of pain and ranges of movement were measured. Function and disability was measured using fifteen instruments, the content of which varied considerably. Function and disability, pain and range of movement (132, 108...

  3. Macro and micro minerals: are frozen fruits a good source?

    Directory of Open Access Journals (Sweden)

    Patricia D.S. Spada

    2010-12-01

    Full Text Available Fruits are rich in minerals, which are essential for a wide variety of metabolic and physiologic processes in the human body. The use of frozen fruits has greatly spread in the last years not only in the preparation of juices, but also as raw material for yogurts, candies, cookies, cakes, ice creams, and children's food. However, up to now there is no data about the mineral profile of frozen fruits. This is the first database to quantify the levels of minerals in 23 samples of frozen fruits, including the most used around the world and some native fruits from the Amazon rainforest in Brazil. Considering the Dietary Reference Intakes, 100g of frozen fruits can provide 0.2 to 2.8% of macro and 2.5 to 100% of microminerals for adults (31-50 years old. Although geographical differences should be considered, these data can help to plan diets and to develop population interventions aiming to prevent chronic diseases.As frutas são ricas em minerais, sendo estes essenciais para uma grande variedade de processos metabólicos e fisiológicos no corpo humano. A utilização de frutas congeladas tem se ampliado nos últimos anos, não só na preparação de sucos, mas também como matéria-prima para iogurtes, doces, biscoitos, bolos, sorvetes e alimentos infantis. No entanto, até o momento não há dados sobre o perfil mineral de frutas congeladas. Este trabalho é o primeiro banco de dados para quantificar os níveis de minerais em 23 amostras de frutas congeladas, bastante consumidas em todo o mundo e de algumas frutas nativas da floresta amazônica, Brasil. Considerando-se as Referências de Ingestão Diárias, 100g de frutas congeladas podem fornecer 0,2-2,8% de macro e de 2,5 a 100% dos mi-crominerais para adultos (31-50 anos. Embora as diferenças geográficas devam ser consideradas, estes dados ajudam para o plano de dietas e desenvolvimento de intervenções junto à população co o objetivo de prevenir doenças crônicas.

  4. Melting Frozen Droplets Using Photo-Thermal Traps

    Science.gov (United States)

    Dash, Susmita; de Ruiter, Jolet; Varanasi, Kripa

    2017-11-01

    Ice buildup is an operational and safety hazard in wind turbines, power lines, and airplanes. While traditional de-icing methods are energy-intensive or environmentally unfriendly, passive anti-icing approach using superhydrophobic surfaces fails under humid conditions, which necessitates development of passive deicing methods. Here, we investigate a passive technique for deicing using a multi-layer surface design that can efficiently absorb and convert the incident solar radiation to heat. The corresponding increase in substrate temperature allows for easy removal of frozen droplets from the surface. We demonstrate the deicing performance of the designed surface both at very low temperatures, and under frost and snow coverage.

  5. Deteksi Kerusakan DNA Spermatozoa Semen Segar dan Semen Beku Sapi Menggunakan Pewarnaan Toluidine Blue (DETECTION OF SPERM DNA DAMAGE INFRESH AND FROZEN SEMEN USING TOLUIDINE BLUE STAINING

    Directory of Open Access Journals (Sweden)

    Langgeng Priyanto

    2015-05-01

    Full Text Available Sperm DNA integrity is very important in the success of fertilization and embryo development.Toluidine blue (TB is a sensitive staining to examine the chromatin structure of sperm. The aim of thisstudy was to detect the sperm DNA damage before and after freezing using TB. Semen was obtained fromeight superior bulls (two Brahman, two Ongole, two Simental and two Limosin belong to Lembang ArtificialInsemination Centre. Semen was collected twice a week using artificial vagina, then was evaluatedmacrocopically and microscopically after collected, including sperm motility, viability, plasma membraneintegrity (MI and acrosome intact (AI, sperm concentration, abnormality and sperm DNA integrity. Thesemen that been used in this study showed the total motility was more than 70%, sperm concentrationwas more than 1000x106, and the sperm abnormality was below 20%. The result showed that the qualityof semen after freezing processed was decreased significantly (P<0.05 on percentage of sperm motility,viability, MI and AI, whereas there was no different on DNA integrity (P>0.05. Sperm DNA integrity offresh and frozen semen were 93.91±4.77% and 92.06 ±2.41% respectively. The decrease of DNA integritywas low (1.84% compared to motility (28.3%, viability (21.6%, MI (14.1%, and AI (11.8%. In conclusion,toluidine blue can be used to detect of DNA damage and the freezing process will not decrease the DNAintegrity.

  6. Establishment and characterization of primary glioblastoma cell lines from fresh and frozen material: a detailed comparison.

    Directory of Open Access Journals (Sweden)

    Christina Susanne Mullins

    Full Text Available BACKGROUND: Development of clinically relevant tumor model systems for glioblastoma multiforme (GBM is important for advancement of basic and translational biology. High molecular heterogeneity of GBM tumors is well recognized, forming the rationale for molecular tests required before administration of several of the novel therapeutics rapidly entering the clinics. One model that has gained wide acceptance is the primary cell culture model. The laborious and time consuming process is rewarded with a relative high success rate (about 60%. We here describe and evaluate a very simple cryopreservation procedure for GBM tissue prior to model establishment that will considerably reduce the logistic complexity. METHODS: Twenty-seven GBM samples collected ad hoc were prepared for primary cell culture freshly from surgery (#1 and after cryopreservation (#2. RESULTS: Take rates after cryopreservation (59% were as satisfactory as from fresh tissue (63%; p = 1.000. We did not observe any relevant molecular or phenotypic differences between cell lines established from fresh or vitally frozen tissue. Further, sensitivity both towards standard chemotherapeutic agents (Temozolomide, BCNU and Vincristine and novel agents like the receptor tyrosine kinase inhibitor Imatinib did not differ. CONCLUSIONS: Our simple cryopreservation procedure facilitates collection, long-time storage and propagation (modeling of clinical GBM specimens (potentially also from distant centers for basic research, (pre- clinical studies of novel therapies and individual response prediction.

  7. Comparative performance of isolation methods using Preston broth, Bolton broth and their modifications for the detection of Campylobacter spp. from naturally contaminated fresh and frozen raw poultry meat.

    Science.gov (United States)

    Seliwiorstow, T; De Zutter, L; Houf, K; Botteldoorn, N; Baré, J; Van Damme, I

    2016-10-03

    The performance of different isolation methods was evaluated for the detection of Campylobacter from naturally contaminated raw poultry meat. Therefore, fresh and frozen poultry meat samples were analysed using the standard procedure (ISO 10272-1:2006), enrichment in Preston broth, and enrichment in modified Bolton broth (supplemented with (i) potassium clavulanate (C-BB), (ii) triclosan (T-BB), (iii) polymyxin B (P-BB)). The enrichment cultures were streaked onto both modified charcoal cefoperazone deoxycholate agar (mCCDA) and RAPID'Campylobacter agar (RCA). Moreover, direct plating on mCCDA and RCA was performed to quantify Campylobacter. In total, 33 out of 59 fresh retail meat samples (55.9%) were Campylobacter positive. For both fresh and frozen poultry meat samples, enrichment in Bolton broth (ISO 10272-1:2006) resulted in a higher number of positive samples than enrichment in Preston broth. Supplementation of Bolton broth with potassium clavulanate (C-BB) and triclosan (T-BB) enhanced the Campylobacter recovery from fresh poultry meat compared to non-supplemented Bolton broth, although the use of C-BB was less applicable than T-BB for Campylobacter recovery from frozen samples. Additionally, the use of RCA resulted in a higher isolation rate compared to mCCDA. The present study demonstrates the impact of culture medium on the recovery of Campylobacter from fresh and frozen naturally contaminated poultry meat samples and can support laboratories in choosing the most appropriate culturing method to detect Campylobacter. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Synovial fluid white cell count and histopathological examination of periprosthetic tissue samples (frozen and permanent sections in the diagnosis of prosthetic knee infection

    Directory of Open Access Journals (Sweden)

    Obada B.

    2017-02-01

    Full Text Available The aim of the study was to determine prospectively the importance of synovial fluid white cell count and intraoperative frozen and permanent sections analysis (number of polymorphonuclear leukocytes per high-power field in the diagnosis of septic total knee arthroplasty. There were studied prospectively 72 patients who needed a revision total knee arthroplasty between 2013-2015. 30 patients were diagnosed with prosthetic joint infection due to high rates of ESR (93% and CRP (90% and preoperative positive culture from aspirated synovial fluid and 42 patients were considered to have aseptic failure according to negative preoperative culture from joint aspirate. For all the patients was analysed synovial fluid white cell count and histopathological aspect of intraoperative frozen and permanent sections of periprosthetic tissue. The results showed a median value of 13800 of sinovial white cells count for infected knee and 92 for noninfected knee. 90% of the patients with joint infection had more than 5 polymorphonuclear leukocytes per high power field on intraoperative frozen sections and 83% on permanent sections. None of the patients from aseptic group had more than 5 polymorphonuclear leukocytes per field on permanent sections. The erythrocyte sedimentation rate and C-reactive protein level can be supplemented with cultures of aspirated joint fluid and fluid white cell count to confirm the diagnosis of periprosthetic infection. When the preoperative diagnosis remain unclear, the histological examination of frozen or permanent sections of periprosthetic tissue with at least 5 polymorphonuclear leukocytes per high power field, is predictive for the presence of infection.

  9. Monitoring of Legionella pneumophila viability after chlorine dioxide treatment using flow cytometry.

    Science.gov (United States)

    Mustapha, Pascale; Epalle, Thibaut; Allegra, Séverine; Girardot, Françoise; Garraud, Olivier; Riffard, Serge

    2015-04-01

    The viability of three Legionella pneumophila strains was monitored after chlorine dioxide (ClO2) treatment using a flow cytometric assay. Suspensions of L. pneumophila cells were submitted to increasing concentrations of ClO2. Culturable cells were still detected when using 4 mg/L, but could no longer be detected after exposure to 6 mg/L of ClO2, although viable but not culturable (VBNC) cells were found after exposure to 4-5 mg/L of ClO2. When testing whether these VBNC were infective, two of the strains were resuscitated after co-culture with Acanthamoeba polyphaga, but neither of them could infect macrophage-like cells. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  10. Evaluation of Giardia duodenalis viability after metronidazole treatment by flow cytometry

    Directory of Open Access Journals (Sweden)

    Joana Barbosa

    2014-12-01

    Full Text Available Giardia duodenalis (syn. lamblia; syn. intestinalis susceptibility testing is not routinely performed because the classical culture methods are very time-consuming and laborious. We developed a novel flow cytometry (FC assay to evaluate the susceptibility of G. duodenalis trophozoites to metronidazole (MTZ. Different concentrations of MTZ were added to cultures of trophozoites (10 5 /mL and the cultures were incubated for different periods. The 50% inhibitory concentration was calculated and propidium iodide (PI was used to quantify the number of dead cells. After treatment, PI-positive trophozoites increased with increasing drug concentration and exposure time. An excellent correlation was found between FC and the classical method. A novel, accurate and reliable method is now available to evaluate G. duodenalis viability.

  11. Motion and Pain Relief Remain 23 Years After Manipulation Under Anesthesia for Frozen Shoulder

    National Research Council Canada - National Science Library

    Vastamäki, Heidi; Vastamäki, Martti

    2013-01-01

    Manipulation under anesthesia (MUA) as treatment for idiopathic frozen shoulder increases motion, provides pain relief, and restores function, but it is unclear whether the improvements persist long...

  12. Free polyunsaturated fatty acids cause taste deterioration of salmon during frozen storage

    DEFF Research Database (Denmark)

    Refsgaard, Hanne; Brockhoff, P.M.B.; Jensen, Benny

    2000-01-01

    Increased intensity of train oil taste, bitterness, and metal taste are the most pronounced sensory changes during frozen storage of salmon (Refsgaard, H. H. F.; Brockhoff, P. B.; Jensen, B. Sensory and Chemical Changes in Farmed Atlantic Salmon (Salmo salar) during Frozen Storage. J. Agric. Food...... of train oil taste, bitterness and metal taste was in the order: DHA > palmitoleic acid > linoleic acid > EPA. Formation of free fatty acids was inhibited by cooking the salmon meat before storage. Furthermore, no changes in phospholipid level were observed during frozen storage. The results suggest...... that enzymatic hydrolysis of neutral lipids plays a major role in the sensory deterioration of salmon during frozen storage....

  13. Evaluation of thermal effects and strain-rate sensitivity in frozen soil

    Directory of Open Access Journals (Sweden)

    Zhu Zhi-Wu

    2014-01-01

    Full Text Available Temperature variation is one important factor that affects the dynamic mechanical properties of frozen soil under impact loading. Thermal damage is a collective phenomenon that can be caused by temperature variation. This paper investigates the effects of thermal damage on strain course. A split Hopkinson pressure bar was employed to investigate the dynamic mechanical characteristics of frozen soil at different temperatures and different strain rates. The stress-strain curves were obtained under impact loading. The compressive strength of frozen soil showed a negative temperature sensitivity and positive strain-rate trend. Specifically, the strength of frozen soil increased with decreasing temperatures and increasing strain rates.

  14. Patients' Attitudes towards the Surplus Frozen Embryos in China

    Directory of Open Access Journals (Sweden)

    Xuan Jin

    2013-01-01

    Full Text Available Background. Assisted reproductive techniques have been used in China for more than 20 years. This study investigates the attitudes of surplus embryo holders towards embryos storage and donation for medical research. Methods. A total of 363 couples who had completed in vitro fertilization (IVF treatment and had already had biological children but who still had frozen embryos in storage were invited to participate. Interviews were conducted by clinics in a narrative style. Results. Family size was the major reason for participants’ (discontinuation of embryo storage; moreover, the moral status of embryos was an important factor for couples choosing embryo storage, while the storage fee was an important factor for couples choosing embryo disposal. Most couples discontinued the storage of their embryos once their children were older than 3 years. In our study, 58.8% of the couples preferred to dispose of surplus embryos rather than donate them to research, citing a lack of information and distrust in science as significant reasons for their decision. Conclusions. Interviews regarding frozen embryos, including patients’ expectations for embryo storage and information to assist them with decisions regarding embryo disposal, are beneficial for policies addressing embryo disposition and embryo donation in China.

  15. Field-scale water balance closure in seasonally frozen conditions

    Directory of Open Access Journals (Sweden)

    X. Pan

    2017-11-01

    Full Text Available Hydrological water balance closure is a simple concept, yet in practice it is uncommon to measure every significant term independently in the field. Here we demonstrate the degree to which the field-scale water balance can be closed using only routine field observations in a seasonally frozen prairie pasture field site in Saskatchewan, Canada. Arrays of snow and soil moisture measurements were combined with a precipitation gauge and flux tower evapotranspiration estimates. We consider three hydrologically distinct periods: the snow accumulation period over the winter, the snowmelt period in spring, and the summer growing season. In each period, we attempt to quantify the residual between net precipitation (precipitation minus evaporation and the change in field-scale storage (snow and soil moisture, while accounting for measurement uncertainties. When the residual is negligible, a simple 1-D water balance with no net drainage is adequate. When the residual is non-negligible, we must find additional processes to explain the result. We identify the hydrological fluxes which confound the 1-D water balance assumptions during different periods of the year, notably blowing snow and frozen soil moisture redistribution during the snow accumulation period, and snowmelt runoff and soil drainage during the melt period. Challenges associated with quantifying these processes, as well as uncertainties in the measurable quantities, caution against the common use of water balance residuals to estimate fluxes and constrain models in such a complex environment.

  16. COAGULATION FACTORS LEVEL IN FRESH FROZEN PLASMA IN RWANDA.

    Science.gov (United States)

    Uwamungu, S; Nyamache, A K; Masaisa, F; Njoki, S K; Abdalah, F; Saibu, K; Ndahiriwe, O; Agwata, D

    2014-02-01

    Objectives: To determine the level of coagulation factors and inherited inhibitors in Fresh Frozen Plasma (FFP) and to evaluate Prothrombin Time and activated partial thrombin time in fresh frozen plasma. Cross-sectional study. Jomo Kenyatta University of Agriculture and Technology in Medical Laboratory Sciences. Eighteen blood bags collected from voluntary blood donors. Coagulation factors and inhibitors levels, Prothrombin Time (PT) and Activated Partial thrombin Time (APTT) remained within the reference range requested by quality assurance regulations after three months of storage. APTT and PT show an increase from baseline to one month then remain constant up to three months, while, Fibrinogen, Factor II, Factor V, Factor VII, Factor X, Von Willbrand Factor, Protein C and Antithrombin decreased from baseline up to three months and then Factor VIII, Factor IX, Factor XI, Factor XII and Protein S, remained constant from baseline up to one month and decreased up to three months. There is good retention of all coagulation factors and inhibitors in plasma produced from whole blood within eight hours of collection, stored at minus 18 degrees C for three months.

  17. Field-scale water balance closure in seasonally frozen conditions

    Science.gov (United States)

    Pan, Xicai; Helgason, Warren; Ireson, Andrew; Wheater, Howard

    2017-11-01

    Hydrological water balance closure is a simple concept, yet in practice it is uncommon to measure every significant term independently in the field. Here we demonstrate the degree to which the field-scale water balance can be closed using only routine field observations in a seasonally frozen prairie pasture field site in Saskatchewan, Canada. Arrays of snow and soil moisture measurements were combined with a precipitation gauge and flux tower evapotranspiration estimates. We consider three hydrologically distinct periods: the snow accumulation period over the winter, the snowmelt period in spring, and the summer growing season. In each period, we attempt to quantify the residual between net precipitation (precipitation minus evaporation) and the change in field-scale storage (snow and soil moisture), while accounting for measurement uncertainties. When the residual is negligible, a simple 1-D water balance with no net drainage is adequate. When the residual is non-negligible, we must find additional processes to explain the result. We identify the hydrological fluxes which confound the 1-D water balance assumptions during different periods of the year, notably blowing snow and frozen soil moisture redistribution during the snow accumulation period, and snowmelt runoff and soil drainage during the melt period. Challenges associated with quantifying these processes, as well as uncertainties in the measurable quantities, caution against the common use of water balance residuals to estimate fluxes and constrain models in such a complex environment.

  18. The concentration of antibiotic in fresh-frozen bone graft.

    Science.gov (United States)

    Chang, Y; Shih, H-N; Chen, D W; Lee, M S; Ueng, S W; Hsieh, P-H

    2010-10-01

    We investigated the antibiotic concentration in fresh-frozen femoral head allografts harvested from two groups of living donors. Ten samples were collected from patients with osteoarthritis of the hip and ten from those with a fracture of the neck of the femur scheduled for primary arthroplasty. Cefazolin (1 g) was administered as a pre-operative prophylactic antibiotic. After storage at -80 degrees C for two weeks the pattern of release of cefazolin from morsellised femoral heads was evaluated by an in vitro broth elution assay using high-performance liquid chromatography. The bioactivity of the bone was further determined with an agar disc diffusion and standardised tube dilution bioassay. The results indicated that the fresh-frozen femoral heads contained cefazolin. The morsellised bone released cefazolin for up to four days. The concentration of cefazolin was significantly higher in the heads from patients with osteoarthritis of the hip than in those with a fracture.Also, in bioassays the bone showed inhibitory effects against bacteria.We concluded that allografts of morsellised bone from the femoral head harvested from patients undergoing arthroplasty of the hip contained cefazolin, which had been administered pre-operatively and they exhibited inhibitory effects against bacteria in vitro.

  19. Rapid Detection of Human Norovirus in Frozen Raspberries.

    Science.gov (United States)

    Summa, Maija; Maunula, Leena

    2017-10-10

    Raspberries have lately caused several human norovirus (HuNoV) outbreaks in Europe. In this study, we developed and evaluated for HuNoV reverse transcription (RT)-PCR detection in frozen raspberries extraction methods that have equal sensitivity but are less time-consuming than widely used methods based on polyethylene glycol (PEG) precipitation and chloroform-butanol purification. One method was applied to stored frozen raspberries linked to previous HuNoV outbreaks and berries on sale. In the virus elution-based Method 1, sparkling water eluted viruses most efficiently from the berries. Method 2, based on direct nucleic acid extraction with minor PEG supplement, yielded the highest number of positive findings (4 out of 9) at low virus concentration level of 100 genome copies HuNoV genogroup II per 25 g raspberries. Both methods showed approximately equal sensitivity to a method including PEG precipitation and chloroform-butanol purification. Two naturally contaminated berry samples linked to HuNoV outbreaks in 2006 and 2009 were still positive for HuNoV genogroup I, but all berry products purchased from a local store remained negative for HuNoV. In conclusion, this study presents two efficient and rapid methods which can be used in urgent HuNoV outbreak investigations, since the results of the virus analysis are available in a few hours.

  20. The Object(s) of Culture: Bruno Latour and the Relationship between Science and Culture.

    Science.gov (United States)

    Doll, William E., Jr.; Petrina, Stephen

    This paper begins by describing the type of newspaper stories about global warming, AIDS vaccines, and frozen embryos that are neither pure science nor pure politics. The paper states that the French sociologist and commentator on cultural trends, Bruno Latour, calls such stories "hybrid articles." It suggests that the purity of the…

  1. Viability of Strongyloides venezuelensis eggs and larvae in vermiculite containing the fungus Duddingtonia flagrans.

    Science.gov (United States)

    Silva, Laryssa Pinheiro Costa; Ferraz, Carolina Magri; Aguiar, Anderson Rocha; Araújo, Jackson Victor; Ribeiro, Steveen Rios; Rossi, Débora Goldner; Mendes, Luanderson Queiroz; Pereira, Fausto Edmundo Lima; Moreira, Narcisa Imaculada Brant; Braga, Fabio Ribeiro

    2017-07-01

    Strongyloidiasis is the most clinically important disease among the infections caused by geohelminths, seeing that this parasite can cause autoinfection. The use of nematophagous fungi like Duddingtonia flagrans, that have predation action on eggs and infecciososas forms of helminths, emerges as an alternative method for environmental control. For this reason, analyzing the viability of larvae and eggs of Strongyloides venezuelensis and the action of Duddingtonia flagrans AC001 in vermiculite, as well as the action of the nematophagous fungi in different growth stages, is important to elaborate and define the best culture conditions that favor the activity of the fungus. Two different growth conditions were applied: both eggs and AC001 fungi were added at the same time to the vermiculite (assay A) and the addition of eggs after the growth of the AC001 fungi in the vermiculite (assay B). To recover the L3 larvae, the Baermann-Moraes method was applied, followed by the counting of L3 dead and alive. At last, it was observed that the vermiculite enriched with organic material is an adequate culture medium not only for the growth of the S. venezuelensis but also for the growth of the D. flagrans fungus, being therefore, a satisfactory culture medium for tests of viability and predatory action of this fungus. It was also observed that the activity of the AC001 fungus is greater when it is growing concomitantly with the eggs, in other words, when it is in the adaptation phase.

  2. Effect of cadmium on cellular viability in two species of microalgae (Scenedesmus sp. and Dunaliella viridis).

    Science.gov (United States)

    Marcano, Letty Beatriz C; Carruyo, Ingrid M; Montiel, Xiomara M; Morales, Carolina B; de Soto, Patricia Moreno

    2009-07-01

    We determined the effect of several concentrations of cadmium (0, 5, 10, and 20 microg/l) on cellular viability in the microalgae Scenedesmus sp. and Dunaliella viridis, by measuring growth at 0, 24, 48, 72, and 96 h and pigment production at 10 days. Algae were obtained from the Nonvascular Plant Laboratory collection, in the Facultad Experimental de Ciencias, Universidad del Zulia, Venezuela. Growth was measured by cellular counting, while pigment content was evaluated using conventional spectrophotometric techniques. Growth of both species decreased in the exposed cultures comparing with the control, but its behavior was similar, because in both control and exposed cultures, its was observed an adaptive phase in the first hours, as well as a growth phase after 72 h. Cadmium concentrations above 10 microg/l produced an adverse effect on pigment production, depending on the concentration and/or exhibition time. However, even though cadmium inhibited growth and pigment production, levels of both parameters indicated cellular viability, demonstrating the adaptability of the algae cultures when they were exposed to the metal.

  3. The effect of ketorolac tromethamine, methylprednisolone, and platelet-rich plasma on human chondrocyte and tenocyte viability.

    Science.gov (United States)

    Beitzel, Knut; McCarthy, Mary Beth; Cote, Mark P; Apostolakos, John; Russell, Ryan P; Bradley, James; ElAttrache, Neal S; Romeo, Antony A; Arciero, Robert A; Mazzocca, Augustus D

    2013-07-01

    The purpose of this study was to evaluate the effect on cell viability of the isolated and combined use of allogeneic platelet-rich plasma (PRP) and ketorolac tromethamine on human chondrocytes and tenocytes in a highly controlled in vitro environment. PRP was produced from 8 subjects. Human chondrocytes (Lonza, Hopkinton, MA) and tenocytes isolated from samples of the long head of the biceps tendons were treated in culture with PRP, ketorolac tromethamine, and methylprednisolone, both alone and in combination. Control samples were treated in media containing 2% or 10% fetal bovine serum (FBS). Cells were exposed for 1 hour. Luminescence assays were obtained to examine cell viability after 24 hours and long-term effects on cell viability after 120 hours. Radioactive thymidine assay was used to measure proliferation after 120 hours. For chondrocytes, cell viability (120 hours) increased significantly with the treatment of PRP alone (43,949 ± 28,104 cells; P investigation into alternative treatment options such as combinations of PRP and ketorolac tromethamine. In vitro evaluation of their effect on cell viability might build a basis for further translational research and clinical application. Copyright © 2013 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.

  4. The market viability of nuclear hydrogen technologies.

    Energy Technology Data Exchange (ETDEWEB)

    Botterud, A.; Conzelmann, G.; Petri, M. C.; Yildiz, B.

    2007-04-06

    The Department of Energy Office of Nuclear Energy is supporting system studies to gain a better understanding of nuclear power's potential role in a hydrogen economy and what hydrogen production technologies show the most promise. This assessment includes identifying commercial hydrogen applications and their requirements, comparing the characteristics of nuclear hydrogen systems to those market requirements, evaluating nuclear hydrogen configuration options within a given market, and identifying the key drivers and thresholds for market viability of nuclear hydrogen options. One of the objectives of the current analysis phase is to determine how nuclear hydrogen technologies could evolve under a number of different futures. The outputs of our work will eventually be used in a larger hydrogen infrastructure and market analysis conducted for DOE-EE using a system-level market simulation tool now underway. This report expands on our previous work by moving beyond simple levelized cost calculations and looking at profitability, risk, and uncertainty from an investor's perspective. We analyze a number of technologies and quantify the value of certain technology and operating characteristics. Our model to assess the profitability of the above technologies is based on Real Options Theory and calculates the discounted profits from investing in each of the production facilities. We use Monte-Carlo simulations to represent the uncertainty in hydrogen and electricity prices. The model computes both the expected value and the distribution of discounted profits from a production plant. We also quantify the value of the option to switch between hydrogen and electricity production in order to maximize investor profits. Uncertainty in electricity and hydrogen prices can be represented with two different stochastic processes: Geometric Brownian Motion (GBM) and Mean Reversion (MR). Our analysis finds that the flexibility to switch between hydrogen and electricity leads

  5. Culture.:'Culture.'

    OpenAIRE

    Thin, Neil; Biswas-Diener, Robert

    2016-01-01

    Although the most visible elements of culture are dress, cuisine and architecture, culture is a highly psychological phenomenon. Culture is a pattern of meaning for understanding how the world works. This knowledge is shared among a group of people and passed from one generation to the next. This module defines culture, addresses methodological issues, and introduces the idea that culture is a process. Understanding cultural processes can help people get along better with others and be more s...

  6. Loss of viability and induction of apoptosis in human keratinocytes exposed to Staphylococcus aureus biofilms in vitro.

    Science.gov (United States)

    Kirker, Kelly R; Secor, Patrick R; James, Garth A; Fleckman, Philip; Olerud, John E; Stewart, Philip S

    2009-01-01

    Bacteria colonizing chronic wounds are believed to exist as polymicrobial, biofilm communities; however, there are few studies demonstrating the role of biofilms in chronic wound pathogenesis. This study establishes a novel method for studying the effect of biofilms on the cell types involved in wound healing. Cocultures of Staphylococcus aureus biofilms and human keratinocytes (HK) were created by initially growing S. aureus biofilms on tissue culture inserts then transferring the inserts to existing HK cultures. Biofilm-conditioned medium (BCM) was prepared by culturing the insert-supported biofilm in cell culture medium. As a control planktonic-conditioned medium (PCM) was also prepared. Biofilm, BCM, and PCM were used in migration, cell viability, and apoptosis assays. Changes in HK morphology were followed by brightfield and confocal microscopy. After only 3 hours exposure to BCM, but not PCM, HK formed dendrite-like extensions and displayed reduced viability. After 9 hours, there was an increase in apoptosis (pPCM-exposed HK all exhibited reduced scratch closure (p< or =0.0001). The results demonstrated that soluble products of both S. aureus planktonic cells and biofilms inhibit scratch closure. Furthermore, S. aureus biofilms significantly reduced HK viability and significantly increased HK apoptosis compared with planktonic S. aureus.

  7. Viability of zebrafish (Danio rerio) ovarian follicles after vitrification in a metal container.

    Science.gov (United States)

    Marques, Lis S; Bos-Mikich, Adriana; Godoy, Leandro C; Silva, Laura A; Maschio, Daniel; Zhang, Tiantian; Streit, Danilo P

    2015-12-01

    Cryopreservation of ovarian tissue has been studied for female germline preservation of farm animals and endangered mammalian species. However, there are relatively few reports on cryopreservation of fish ovarian tissue and especially using vitrification approach. Previous studies of our group has shown that the use of a metal container for the cryopreservation of bovine ovarian fragments results in good primordial and primary follicle morphological integrity after vitrification. The aim of this study was to assess the viability and in vitro development of zebrafish follicles after vitrification of fragmented or whole ovaries using the same metal container. In Experiment 1, we tested the follicular viability of five developmental stages following vitrification in four vitrification solutions using fluorescein diacetate and propidium iodide fluorescent probes. These results showed that the highest viability rates were obtained with immature follicles (Stage I) and VS1 (1.5 M methanol + 4.5 M propylene glycol). In Experiment 2, we used VS1 to vitrify different types of ovarian tissue (fragments or whole ovaries) in two different carriers (plastic cryotube or metal container). In this experiment, Stage I follicle survival was assessed following vitrification by vital staining after 24 h in vitro culture. Follicular morphology was analyzed by light microscopy after vitrification. Data showed that the immature follicles morphology was well preserved after cryopreservation. Follicular survival rate was higher (P vitrification devices were compared. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. 3D Printed Vascular Networks Enhance Viability in High-Volume Perfusion Bioreactor.

    Science.gov (United States)

    Ball, Owen; Nguyen, Bao-Ngoc B; Placone, Jesse K; Fisher, John P

    2016-12-01

    There is a significant clinical need for engineered bone graft substitutes that can quickly, effectively, and safely repair large segmental bone defects. One emerging field of interest involves the growth of engineered bone tissue in vitro within bioreactors, the most promising of which are perfusion bioreactors. Using bioreactor systems, tissue engineered bone constructs can be fabricated in vitro. However, these engineered constructs lack inherent vasculature and once implanted, quickly develop a necrotic core, where no nutrient exchange occurs. Here, we utilized COMSOL modeling to predict oxygen diffusion gradients throughout aggregated alginate constructs, which allowed for the computer-aided design of printable vascular networks, compatible with any large tissue engineered construct cultured in a perfusion bioreactor. We investigated the effect of 3D printed macroscale vascular networks with various porosities on the viability of human mesenchymal stem cells in vitro, using both gas-permeable, and non-gas permeable bioreactor growth chamber walls. Through the use of 3D printed vascular structures in conjunction with a tubular perfusion system bioreactor, cell viability was found to increase by as much as 50% in the core of these constructs, with in silico modeling predicting construct viability at steady state.

  9. Puget Sound steelhead life cycle model analyses - Population Viability Analysis

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This research was initiated by the Puget Sound Steelhead Technical Recovery Team to develop viability criteria for threatened Puget Sound steelhead and to support...

  10. Cadmium affects the mitochondrial viability and the acid soluble ...

    African Journals Online (AJOL)

    Cadmium affects the mitochondrial viability and the acid soluble thiols concentration in liver, kidney, heart and gills of Ancistrus brevifilis (Eigenmann, 1920). P Velasquez-Vottelerd, Y Anton, R Salazar-Lugo ...

  11. STUDY ON POLLEN VIABILITY AS BIOINDICATOR OF AIR QUALITY

    Directory of Open Access Journals (Sweden)

    Florentina ŞTEFLEA

    2012-01-01

    Full Text Available The aim of this research is to estimate the relationship between pollen viability and atmospheric pollution (in polluted and non-polluted conditions. The study was carried out in the city of Timisoara. Two areas, with different intensity of road traffic (very high and absent but all characterized by the presence of the same plant species, were selected. The pollen of herbaceous spontaneous species, arboreal species and a shrub species was used (Robinia pseudacacia, Aesculus x carnea, Catalpa bignonioides, Albizzia julibrissin, Rosa canina, Sambucus nigra, Malva neglecta, Ranunculus acer, Trifolium repens, Cichorium intybus. The pollen of these species was treated with TTC (2, 3, 5 Tryphenil-Tetrazolium-Chloride staining solution and viability was then estimated by light microscopy. The results of the mean pollen viability percentage of the examined species are reported. Pollen viability of herbaceous plants is significantly different between the two environments.

  12. Maintaining yeast viability in continuous primary beer fermentation

    National Research Council Canada - National Science Library

    Pires, Eduardo J; Teixeira, José A; Brányik, Tomás; Côrte‐Real, Manuela; Vicente, António A

    2014-01-01

    .... This work was aimed at solving one of the most relevant obstacles to implementing ICT on a large scale in beer fermentations, namely the control of biomass and the maintenance of cell viability in a gas‐lift bioreactor...

  13. Desiccation-induced changes in viability, lipid peroxidation and ...

    African Journals Online (AJOL)

    user

    2012-05-31

    Hendry et al., 1992) and A. saccharinum (Pukacka and Ratajczak, 2006) and intermediate seeds like Azadirachta indica (Varghese and. Naithani, 2002), Coffea Arabica (Dussert et al., 2006), indicating that loss of seed viability ...

  14. Femtosecond optical transfection of cells:viability and efficiency

    National Research Council Canada - National Science Library

    D. Stevenson; B. Agate; X. Tsampoula; P. Fischer; C. T. A. Brown; W. Sibbett; A. Riches; F. Gunn-Moore; K. Dholakia

    2006-01-01

    .... However, there remains no study into the true efficiency of this procedure. Here, we present a detailed analysis of transfection efficiency and cell viability for femtosecond optical transfection using a titanium sapphire laser at 800 nm...

  15. Approximate viability for nonlinear evolution inclusions with application to controllability

    Directory of Open Access Journals (Sweden)

    Omar Benniche

    2016-12-01

    Full Text Available We investigate approximate viability for a graph with respect to fully nonlinear quasi-autonomous evolution inclusions. As application, an approximate null controllability result is given.

  16. Assessment of frozen storage duration effect on quality characteristics of various horse muscles

    Directory of Open Access Journals (Sweden)

    Pil Nam Seong

    2017-12-01

    Full Text Available Objective The study aimed at assessing the effects of frozen storage duration on quality characteristics, lipid oxidation and sensory quality of various horse muscles. Methods Five representative muscles: longissimus dorsi (LD, gluteus medius (GM, semimembranosus (SM, biceps femoris (BF, and triceps brachii (TB at 24 h post-mortem obtained from 28-mo-old Jeju female breed horses (n = 8 were used in the present investigation. The muscles were vacuum-packaged and frozen at −20°C for 120, 240, and 360 days. All the samples were analyzed for thawing and cooking losses, pH, Warner–Bratzler shear forces (WBSF, color traits, total volatile basic nitrogen (TVBN, thiobarbituric acid reactive substances (TBARS and sensory traits. The muscle samples analyzed on day 0 of frozen storage (fresh, non-frozen were used for comparison. Results Results revealed that thawing and cooking losses significantly (p<0.05 increased in all the muscles after 120 days and then remained unchanged up to 360 days of frozen storage. The TBARS and TVBN contents significantly increased as increasing frozen storage time up to 360 days (p<0.05. While, significant decreases in WBSF values were observed for all the muscles with increased frozen storage time (p<0.05. Frozen storage variously affected the color traits of the muscles for instance; the redness of LD, GM, and BF muscles showed a decreasing tendency during frozen storage while it was not changed in TB and SM muscles. Furthermore, the frozen storage did not produce detrimental effects on sensory quality as it did not cause flavor and juiciness defects whereas it partially improved the tenderness of all the muscles studied. Conclusion Based on the results obtained from our work, it is concluded that frozen storage could be applied to increase the long-term shelf life of horsemeat while still retaining its sensory quality.

  17. Evaluation of goat milk as storage media to preserve viability of human periodontal ligament cells in vitro.

    Science.gov (United States)

    Ulusoy, Ayça Tuba; Kalyoncuoglu, Elif; Kaya, Senay; Cehreli, Zafer Cavit

    2016-08-01

    The purpose of this study was to evaluate the effectiveness of goat milk as a storage media for maintenance of periodontal ligament (PDL) cell viability of avulsed teeth and compare it with commonly used and/or investigated storage media. PDL cells were obtained from the root surface of healthy premolars and were cultured in Eagle's maintenance medium (EMM). Cell cultures were treated with the following storage media: tap water (negative control); EMM (positive control); Hank's balanced salt solution; ultra high temperature (UHT) long-shelf-life lactose-free cow milk; UHT long-shelf-life whole cow milk; UHT long-shelf-life skimmed cow milk; UHT long-shelf-life soy milk; UHT long-shelf-life goat milk, UHT long-shelf-life follow on milk with probiotic, 20% propolis, and egg white. Culture plates were incubated with experimental media at 20°C for 1, 3, 6, 12, and 24 h. PDL cell viability was assessed by tetrazolium salt-based colorimetric (MTT) assay at each test period. One-way anova was used to evaluate the effects of storage solutions at each time point, followed by post hoc Duncan's multiple comparison test (P = 0.05). A dendrogram was constructed to show the arrangement of hierarchical clustering. Goat milk displayed the highest capacity to maintain cell viability at all test intervals (P milk with the probiotic showed the lowest time-dependent PDL cell viability among all test media (P milks, HBSS performed significantly less effectively in maintaining PDL cell viability during the entire test period (P milk can be recommended as a suitable storage medium for avulsed teeth. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. Effect of photobiomodulation on viability and proliferation of stem cells from exfoliated deciduous teeth under different nutritional conditions

    Science.gov (United States)

    Morato de Souza, Letícia; Guilherme Roque Rinco, Ugo; Aparecida Tavares Aguiar, Daniela; Aparecido de Almeida Junior, Luciano; Cosme-Silva, Leopoldo; Marchini Oliveira, Thais; Teixeira Marques, Nádia Carolina; Thiemy Sakai, Vivien

    2018-02-01

    This study aimed to evaluate the effect of different doses of low-level laser irradiation on the viability and proliferation of stem cells from exfoliated deciduous teeth (SHED) cultured under nutritional deficit (cellular stress) or regular nutritional conditions. SHED underwent irradiation by a red laser between 1.2 and 6.2 J cm‑2. Prior to the irradiation, all groups received culture medium (MEMα, Eagle’s minimum essential medium alpha modification) supplemented with 1% of fetal bovine serum (FBS) for 1 h. After the irradiation, cells received MEMα supplemented with 10% of FBS (regular nutrition) or 1% of FBS (nutritional deficit). Cell viability and proliferation were respectively determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet assays 6 and 24 h after irradiation (P  nutritional deficit showed lower viability and proliferation after 1.2 J cm‑2 irradiation. All of the irradiated groups revealed significantly higher viability and proliferation in SHED maintained under nutritional deficit than in regular nutritional conditions, except in the 3.7 and 6.2 J cm‑2 groups by MTT assay. In the crystal violet assay, SHED irradiated with 1.2 J cm‑2 showed no difference between the different nutritional conditions. Decrease of FBS concentration in the culture medium seems to enhance the sensitivity of SHED to the effects of photobiomodulation therapy. Nutritional stress conditions improved cell viability and proliferation of SHED after laser irradiation, except for 1.2 J cm‑2.

  19. Effect of formulation on the quality of frozen bread dough

    Directory of Open Access Journals (Sweden)

    Myriam M. Salas-Mellado

    2003-06-01

    Full Text Available The main objective of this work was to determine the influence of formulation on the stability of bread dough during frozen storage. Bread doughs containing gluten and trehalose were submitted to mechanical freezing at -30° C and stored frozen for 45 days. Two types of instant yeast were tested: (A for sweet doughs and (B for savoury doughs. Specific volume was significantly affected by the yeast type, type A showing better effect than type B. Frozen storage of the doughs negatively affected the specific volume, crumb hardness and technological score of the bread. The addition of 10% trehalose had a beneficial effect on the cell survival rate for both the yeasts.O principal objetivo deste trabalho foi determinar a influência da formulação na estabilidade da massa de pão durante a estocagem congelada. As massas para pão contendo glúten Vital e trealose foram submetidas a congelamento mecânico de -30° C e estocadas na forma congelada por 45 dias. Foram testados dois tipos de leveduras instantâneas: (A para massas doces, e (B para massas salgadas. Foram avaliados na massa, os efeitos da formulação, tipo de levedura e a estocagem congelada, pelas características de extensibilidade e a contagem de leveduras viáveis, e no pão, pelo volume específico, a dureza do miolo e as características tecnológicas. Somente o volume específico foi influenciado pelo tipo de levedura, mostrando o tipo A um melhor efeito do que o tipo B. A estocagem congelada influenciou negativamente o volume específico, a dureza do miolo e a pontuação tecnológica do pão. A adição de 10% de trealose mostrou um efeito benéfico na taxa de sobrevivência celular para ambos tipos de leveduras.

  20. l-Carnitine in rooster semen cryopreservation: Flow cytometric, biochemical and motion findings for frozen-thawed sperm.

    Science.gov (United States)

    Fattah, A; Sharafi, M; Masoudi, R; Shahverdi, A; Esmaeili, V; Najafi, A

    2017-02-01

    Rooster semen cryopreservation is not efficient for artificial insemination in breeder flocks. l-Carnitine (LC) has been evaluated for effectiveness in cryopreservation media on the characteristics of rooster sperm after freeze-thawing. Motility characteristics, membrane functionality, abnormal morphology, apoptotic like changes, mitochondria activity and lipid peroxidation of rooster sperms were assessed after freeze-thawing with different concentrations of LC in Beltsville medium. Semen samples were collected from 12 roosters, twice a week, and diluted in the extenders that contained different concentrations of LC. Supplementation of Beltsevile with 1 and 2 mM LC was found to result in higher total motility (68.2± 1.7% and 69.1± 1.7%, respectively), progressive motility (28.4± 1.6%, 29.8± 1.6%), membrane functionality (76.2± 1.9% and 75.9± 1.9%), viability (58.2 ± 1.1%, 59.1 ± 1.1%) and lower significant of lipid peroxidation (2.53 ± 0.08 nmol/ml, 2.49 ± 0.08 nmol/ml) compared to control group containing no LC. Lower motility, progressive motility, and viability were observed in frozen-thawed sperm in extender containing 8 mM LC (35.8± 1.7%, 9.6± 1.2% and 27.1 ± 1.2%, respectively) compared to control. Morphology and mitochondrial activity were not affected by different concentrations of LC. Our results showed that supplementation of Beltsville extender with 1 and 2 mM LC significantly improved the quality of rooster sperm quality after freeze-thawing. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Testing a dual-fluorescence assay to monitor the viability of filamentous cyanobacteria.

    Science.gov (United States)

    Johnson, Tylor J; Hildreth, Michael B; Gu, Liping; Zhou, Ruanbao; Gibbons, William R

    2015-06-01

    Filamentous cyanobacteria are currently being engineered to produce long-chain organic compounds, including 3rd generation biofuels. Because of their filamentous morphology, standard methods to quantify viability (e.g., plate counts) are not possible. This study investigated a dual-fluorescence assay based upon the LIVE/DEAD® BacLight™ Bacterial Viability Kit to quantify the percent viability of filamentous cyanobacteria using a microplate reader in a high throughput 96-well plate format. The manufacturer's protocol calls for an optical density normalization step to equalize the numbers of viable and non-viable cells used to generate calibration curves. Unfortunately, the isopropanol treatment used to generate non-viable cells released a blue pigment that altered absorbance readings of the non-viable cell solution, resulting in an inaccurate calibration curve. Thus we omitted this optical density normalization step, and carefully divided cell cultures into two equal fractions before the isopropanol treatment. While the resulting calibration curves had relatively high correlation coefficients, their use in various experiments resulted in viability estimates ranging from below 0% to far above 100%. We traced this to the apparent inaccuracy of the propidium iodide (PI) dye that was to stain only non-viable cells. Through further analysis via microplate reader, as well as confocal and wide-field epi-fluorescence microscopy, we observed non-specific binding of PI in viable filamentous cyanobacteria. While PI will not work for filamentous cyanobacteria, it is possible that other fluorochrome dyes could be used to selectively stain non-viable cells. This will be essential in future studies for screening mutants and optimizing photobioreactor system performance for filamentous cyanobacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Impact of ZnO and Ag Nanoparticles on Bacterial Growth and Viability

    Science.gov (United States)

    Olson, M. S.; Digiovanni, K. A.

    2007-12-01

    Hundreds of consumer products containing nanomaterials are currently available in the U.S., including computers, clothing, cosmetics, sports equipment, medical devices and product packaging. Metallic nanoparticles can be embedded in or coated on product surfaces to provide antimicrobial, deodorizing, and stain- resistant properties. Although these products have the potential to provide significant benefit to the user, the impact of these products on the environment remains largely unknown. The purpose of this project is to study the effect of metallic nanoparticles released to the environment on bacterial growth and viability. Inhibition of bacterial growth was tested by adding doses of suspended ZnO and Ag nanoparticles into luria broth prior to inoculation of Escherichia coli cells. ZnO particles (approximately 40 nm) were obtained commercially and Ag particles (12-14 nm) were fabricated by reduction of silver nitrate with sodium borohydride. Toxicity assays were performed to test the viability of E. coli cells exposed to both ZnO and Ag nanoparticles using the LIVE/DEAD BacLight bacterial viability kit (Invitrogen). Live cells stain green whereas cells with compromised membranes that are considered dead or dying stain red. Cells were first grown, stained, and exposed to varying doses of metallic nanoparticles, and then bacterial viability was measured hourly using fluorescence microscopy. Results indicate that both ZnO and Ag nanoparticles inhibit the growth of E. coli in liquid media. Preliminary results from toxicity assays confirm the toxic effect of ZnO and Ag nanoparticles on active cell cultures. Calculated death rates resulting from analyses of toxicity studies will be presented.

  3. Effects of voluntary exercise on the viability, proliferation and BDNF levels of bone marrow stromal cells in rat pups born from morphine- dependent mothers during pregnancy.

    Science.gov (United States)

    Haydari, Sakineh; Safari, Manouchehr; Zarbakhsh, Sam; Bandegi, Ahmad Reza; Miladi-Gorji, Hossein

    2016-11-10

    This study was designed to investigate whether free access to a running wheel during pregnancy in morphine-dependent mothers would influence the viability, proliferation and BDNF levels of bone marrow stromal cells in rat pups. Pregnant rats were made dependent by chronic administration of morphine in drinking water simultaneously with free access to a running wheel. Male pups are weaned at 21days of birth and their bones marrows were aspirated from the femurs and tibias and also the bone marrow stromal cells (BMSCs) cultured. MTT assay was used to determine cell viability and proliferation rate. The level of BDNF was measured in the supernant of BMSCs culture by ELISA. The sedentary morphine-dependent mothers' pups showed a significant increase in the percentage cell viability and proliferation rate and also a significant decrease in the BDNF protein levels in BMSCs. The rat pups borne from exercising the control and morphine-dependent mothers exhibited an increase in the percentage viability, proliferation rate and BDNF levels of the BMSCs. This study showed that maternal exercise during pregnancy in morphine-dependent and non-dependent mothers, with increasing of BDNF levels increased the proliferation and viability of BMSCs in the rat pups. Also, chronic administration of morphine during pregnancy was able to increase the proliferation and viability of BMSCs in the rat pups. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  4. Comparative analysis of activity of coagulation Factors V and VIII and level of fibrinogen in fresh frozen plasma and frozen plasma

    Directory of Open Access Journals (Sweden)

    Mitu Dogra

    2015-01-01

    Full Text Available Background: The aim of this study was to analyse and compare the activity of factor V, VIII and fibrinogen level in fresh frozen plasma and frozen plasma frozen after 8 hrs but within 24 hours after phlebotomy. Materials and Methods: Fresh frozen plasma separated from whole blood within 8 hours was compared with plasma separated within 24 hours after phlebotomy in terms of coagulation factors V and VIII and level of fibrinogen by standard methods using semi automated coagulometer sysmex CA50. Results: Longer storage of whole blood before processing resulted in significant decrease (18.4% in activity of factor VIII but the fall in activity of factor V (6.52% or level of fibrinogen (1.81% was not significant. Discussion: These data suggest that there is good retention of coagulation factors in both types of plasma. Although there is significant fall in activity of factor VIII, but it is an acute phase reactant and raised in most of the diseases so it is suggested that frozen plasma would be an acceptable product for most patients requiring fresh frozen plasma.

  5. 78 FR 35643 - Frozen Warmwater Shrimp from China, Ecuador, India, Indonesia, Malaysia, Thailand, and Vietnam...

    Science.gov (United States)

    2013-06-13

    ... COMMISSION Frozen Warmwater Shrimp from China, Ecuador, India, Indonesia, Malaysia, Thailand, and Vietnam..., Malaysia, Thailand, and Vietnam of frozen warmwater shrimp, provided for in subheadings 0306.17.00, 1605.21... found to be subsidized by the Governments of China, India, Malaysia, Thailand, and Vietnam and that are...

  6. 78 FR 764 - Frozen Warmwater Shrimp From China, Ecuador, India, Indonesia, Malaysia, Thailand, and Vietnam...

    Science.gov (United States)

    2013-01-04

    ... COMMISSION Frozen Warmwater Shrimp From China, Ecuador, India, Indonesia, Malaysia, Thailand, and Vietnam..., Malaysia, Thailand, and Vietnam of frozen warmwater shrimp, provided for in subheadings 0306.17.00, 1605.21... subsidized by the Governments of China, Ecuador, India, Indonesia, Malaysia, Thailand, and Vietnam. Unless...

  7. 78 FR 64009 - Frozen Warmwater Shrimp From China, Ecuador, India, Malaysia, and Vietnam

    Science.gov (United States)

    2013-10-25

    ... COMMISSION Frozen Warmwater Shrimp From China, Ecuador, India, Malaysia, and Vietnam Determinations On the... imports from China, Ecuador, India, Malaysia, and Vietnam of frozen warmwater shrimp, provided for in... China, Ecuador, India, Malaysia, and Vietnam.\\2\\ \\1\\ The record is defined in sec. 207.2(f) of the...

  8. Retention of coagulation factors in plasma frozen after extended holding at 1-6 degrees C

    NARCIS (Netherlands)

    Smith, JF; Ness, PM; Moroff, G; Luban, NLC

    2000-01-01

    Background and Objectives: The ability to use plasma, isolated from units of whole blood and frozen within 24 h of phlebotomy, as a substitute for plasma frozen within 8 h of phlebotomy would have several advantages for blood centers. It should provide increased flexibility pertaining to the

  9. Using Disney's "Frozen" to Motivate Mathematics: Bringing Fractals into the Classroom

    Science.gov (United States)

    Piatek-Jimenez, Katrina; Phelps, Christine M.

    2016-01-01

    The movie "Frozen" took the world by storm and this global popularity of the movie and its music can be harnessed by teachers of mathematics. This article builds on the "frozen fractal" lyric from "Let It Go" to incorporate fractal geometry into primary mathematics classrooms.

  10. 77 FR 73424 - Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Partial Rescission of...

    Science.gov (United States)

    2012-12-10

    ... International Trade Administration Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Partial Rescission of Antidumping Duty Administrative Review AGENCY: Import Administration, International Trade... administrative review of the antidumping duty order on certain frozen fish fillets (``fish fillets'') from the...

  11. 78 FR 59915 - Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Initiation of Antidumping...

    Science.gov (United States)

    2013-09-30

    ... International Trade Administration Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Initiation of Antidumping Duty New Shipper Review; 2012-2013 AGENCY: Import Administration, International...'') of the antidumping duty (``AD'') order on certain frozen fish fillets (``fish fillets'') from the...

  12. 78 FR 18957 - Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Initiation of Antidumping...

    Science.gov (United States)

    2013-03-28

    ... International Trade Administration Certain Frozen Fish Fillets From the Socialist Republic of Vietnam: Initiation of Antidumping Duty New Shipper Review; 2012-2013 AGENCY: Import Administration, International...'') of the antidumping duty (``AD'') order on certain frozen fish fillets (``fish fillets'') from the...

  13. Stability after thawing of RBCs frozen with the high- and low-glycerol method

    NARCIS (Netherlands)

    Lelkens, Charles C. M.; Noorman, Femke; Koning, Jack G.; Truijens-de Lange, Rosa; Stekkinger, Perry S.; Bakker, Joa C.; Lagerberg, Johan W. M.; Brand, Anneke; Verhoeven, Arthur J.

    2003-01-01

    BACKGROUND: RBCs can be frozen with either the high-glycerol method (HGM) or the low-glycerol method (LGM). To date, the use of frozen RBCs is hampered by a 24-hour outdating period after thawing. A closed washing system (ACP 215) may solve this problem. STUDY DESIGN AND METHODS: We compared the

  14. 49 CFR 173.229 - Chloric acid solution or chlorine dioxide hydrate, frozen.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Chloric acid solution or chlorine dioxide hydrate, frozen. 173.229 Section 173.229 Transportation Other Regulations Relating to Transportation PIPELINE AND... Than Class 1 and Class 7 § 173.229 Chloric acid solution or chlorine dioxide hydrate, frozen. When the...

  15. The Use of Frozen Section in the Early Diagnosis of Acute Invasive Fungal Sinusitis.

    Science.gov (United States)

    Melancon, C Claire; Clinger, John D

    2017-08-01

    Objectives Acute invasive fungal sinusitis (AIFS) remains a significant cause of morbidity and mortality in the immunocompromised patient population. Early diagnosis is key to improving patient outcomes. Frozen section biopsies have been shown to decrease time to diagnosis when compared with permanent pathology. However, its accuracy has not been adequately described in the literature, specifically in regard to AIFS. The aim of this study is to evaluate the statistical diagnostic accuracy of frozen sections and to review the etiology, clinical presentation, and current diagnostic protocols in management of AIFS. Study Design Case series with chart review. Setting Tertiary referral center. Subjects and Methods Retrospective review included 67 pathologic records in Co-Path, with search criteria including invasive fungal, clinical history, frozen section, and final diagnosis between the dates of 2006 and 2015. Results Sixty-seven cases were reviewed per the search criteria in Co-Path. Of these, 31 met further criteria of having had frozen section analysis. Variables such as sensitivity, specificity, positive predictive value, and negative predictive value were assessed. All 21 positive frozen sections correlated with positive permanent pathology, giving a positive predictive value of 100%. Frozen section biopsies were 87.5% sensitive and 100% specific. Conclusion Early diagnosis of AIFS has been shown to decrease morbidity and mortality. Frozen section biopsies remain key in obtaining an early diagnosis among patients with a high clinical suspicion for invasive fungal sinusitis. Frozen section biopsies positive for invasive fungal pathology were universally consistent with definitive diagnosis.

  16. 75 FR 22424 - Frozen Warmwater Shrimp From Brazil, China, India, Thailand, and Vietnam

    Science.gov (United States)

    2010-04-28

    ... COMMISSION Frozen Warmwater Shrimp From Brazil, China, India, Thailand, and Vietnam AGENCY: United States... Vietnam. SUMMARY: The Commission hereby gives notice that it will proceed with full reviews pursuant to... antidumping duty orders on frozen warmwater shrimp from Brazil, China, India, Thailand, and Vietnam would be...

  17. 75 FR 1078 - Frozen Warmwater Shrimp From Brazil, China, India, Thailand, and Vietnam

    Science.gov (United States)

    2010-01-08

    ... (Review)] Frozen Warmwater Shrimp From Brazil, China, India, Thailand, and Vietnam AGENCY: United States... on frozen warmwater shrimp from Brazil, China, India, Thailand, and Vietnam. SUMMARY: The Commission... warmwater shrimp from Brazil, China, India, Thailand, and Vietnam would be likely to lead to continuation or...

  18. 77 FR 73619 - Final Results of Antidumping Duty Changed Circumstances Review: Certain Frozen Warmwater Shrimp...

    Science.gov (United States)

    2012-12-11

    ... Frozen) is the successor-in-interest to Apex Exports (Apex) for purposes of determining antidumping duty... (Penaeus vannemei), banana prawn (Penaeus merguiensis), fleshy prawn (Penaeus chinensis), giant river prawn... produced and/or exported by Apex Frozen and entered, or withdrawn from warehouse, for consumption on or...

  19. Seismic performance and design of bridge foundations in liquefiable ground with a frozen crust : [summary].

    Science.gov (United States)

    2012-12-01

    Two major earthquakes in Alaska, namely 1964 Great Alaskan earthquake and 2002 Denali earthquake, occurred in winter season when the ground crust was frozen. : None of the then-existing foundation types was able to withstand the force from the frozen...

  20. Seismic performance and design of bridge foundations in liquefiable ground with a frozen crust.

    Science.gov (United States)

    2012-12-01

    Two major earthquakes in Alaska, namely 1964 Great Alaskan earthquake and 2002 Denali earthquake, occurred in winter season when the ground crust was frozen. None of the then-existing foundation types was able to withstand the force from the frozen c...

  1. 9 CFR 327.21 - Inspection procedures for chilled fresh and frozen boneless manufacturing meat.

    Science.gov (United States)

    2010-01-01

    ... fresh and frozen boneless manufacturing meat. 327.21 Section 327.21 Animals and Animal Products FOOD... MEAT AND POULTRY PRODUCTS INSPECTION AND VOLUNTARY INSPECTION AND CERTIFICATION IMPORTED PRODUCTS § 327.21 Inspection procedures for chilled fresh and frozen boneless manufacturing meat. (a) Definitions...

  2. 75 FR 43141 - United States Standards for Grades of Frozen Vegetables

    Science.gov (United States)

    2010-07-23

    ... Agricultural Marketing Service United States Standards for Grades of Frozen Vegetables AGENCY: Agricultural... on the possible revisions to eighteen U.S. grade standards for frozen vegetables issued on or before July 22, 1985. AMS annually reviews its processed fruit and vegetable grade standards for suitability...

  3. Unsaturated Frozen Soil Permeability Coefficient Test and Analysis Under the Condition of Variable Porosity

    Directory of Open Access Journals (Sweden)

    GAO Yu

    2017-06-01

    Full Text Available Frozen soil permeability is one of the key issues in the process of frozen soil water properties research due to special hydrological phenomenon caused by the change of frozen soil water properties. In order to study the effect of ice content and dry density on non - closed porosity and permeability of unsaturated frozen soils, choose ice content ( 2% ,4% ,6% ,8% ,10% ,12% ,14% ,16% ,18% ,20% and dry sample density ( 1. 4 × 103 kg /m3 ,1. 5 × 103 kg /m3 and 1. 6 × 103 kg /m3 as a single variable factor to measure frozen soil permeability coefficient. The water level test method was used to determine the permeability coefficient of frozen soil,and the experimental study on the permeability coefficient of unsaturated frozen soil was carried out. The results show that the permeability coefficient and the unclosed porosity of the unsaturated frozen soil are smaller with the increase of the ice content and dry sample density,and the range is 0. 19 - 0. 714 and 8. 55 - 19. 18 m / d. When the ice content is 20% and the dry density of the sample is 1. 6 × 103 kg /m3 ,the minimum unsaturated permafrost permeability coefficient is 8. 55 m / d and the minimum unclosed porosity is 0. 19.

  4. Researches on the Constitutive Models of Artificial Frozen Silt in Underground Engineering

    Directory of Open Access Journals (Sweden)

    Yugui Yang

    2014-01-01

    Full Text Available The researches on the mechanical characteristic and constitutive models of frozen soil have important meanings in structural design of deep frozen soil wall. In the present study, the triaxial compression and creep tests have been carried out, and the mechanical characteristic of frozen silt is obtained. The experiment results show that the deformation characteristic of frozen silt is related to confining pressure under conventional triaxial compression condition. The frozen silt presents strain softening in shear process; with increase of confining pressure, the strain softening characteristic gradually decreases. The creep curves of frozen silt present the decaying and the stable creep stages under low stress level; however, under high stress level, once the strain increases to a critical value, the creep strain velocity gradually increases and the specimen quickly happens to destroy. To reproduce the deformation behavior, the disturbed state elastoplastic and new creep constitutive models of frozen silt are developed. The comparisons between experimental results and calculated results from constitutive models show that the proposed constitutive models could describe the conventional triaxial compression and creep deformation behaviors of frozen silt.

  5. 75 FR 49460 - Administrative Review of Certain Frozen Warmwater Shrimp From the People's Republic of China...

    Science.gov (United States)

    2010-08-13

    ... International Trade Administration Administrative Review of Certain Frozen Warmwater Shrimp From the People's... Administrative Review of Frozen Warmwater Shrimp from the People's Republic of China: Issues and Decision... revised the wage rate methodology and the surrogate values for shrimp larvae, diesel fuel, shrimp waste...

  6. Effect of stone content on water flow velocity over Loess slope: Frozen soil

    Science.gov (United States)

    Ban, Yunyun; Lei, Tingwu; Feng, Ren; Qian, Dengfeng

    2017-11-01

    Soils in high-altitude or -latitude regions are commonly rich in stone fragments, which are frequently frozen. The hydrodynamics of water flow over frozen, stony slopes must be investigated to understand soil erosion and sediment transportation. The objective of this laboratory experiments was to measure water flow velocity over frozen slopes with different stone contents by using electrolyte trace method. The experiments were performed under slope gradients of 5°, 10°, 15°, and 20°; flow discharge rates of 1, 2, 4, and 8 L/min; and stone contents of 0%, 10%, 20%, and 50% on mass basis. Nine equidistant sensors were used to measure flow velocity along flume from the top of the slope. Results indicated that stone content significantly affected flow velocity under increasing slope gradient. The increase in stone content rapidly reduced the flow velocity. The flow velocities over frozen slopes were 1.21 to 1.30 times of those over non-frozen slopes under different slope gradients and flow rates. When the stone content increased from 0% to 20%, proportions gradually decreased from 52% to 25% and 13%. Additionally, flow velocities over frozen and non-frozen soil slopes became gradually similar with increasing stone content. This study will help elucidate the hydrodynamics, soil erosion, and sediment transport behaviors of frozen or partially unfrozen hillslopes with different stone contents.

  7. Estimation of organelle water fractions from frozen-dried cryosections.

    Science.gov (United States)

    von Zglinicki, T; Bimmler, M; Krause, W

    1987-04-01

    Local dry mass or water fractions can be measured on frozen-dried cryosections assuming constant section thickness in the hydrated state and no net water movements and no differential shrinkage during freezing and drying. These assumptions have been tested on a model consisting of isolated rat liver mitochondria in an albumin matrix with a concentration similar to the dry mass concentration of the cytoplasm. The dry mass concentrations of mitochondria before freezing as measured by interference microscopy and after freezing and freeze-drying of the sections as measured by X-ray microanalysis and scanning microdensitometry are shown to be equal as long as the ice crystals in the medium are smaller than about 100 nm. It is concluded, therefore, that the above-mentioned assumptions could also hold for the cryopreparation of cells and tissues.

  8. Techniques for optimizing nanotips derived from frozen taylor cones

    Science.gov (United States)

    Hirsch, Gregory

    2017-12-05

    Optimization techniques are disclosed for producing sharp and stable tips/nanotips relying on liquid Taylor cones created from electrically conductive materials with high melting points. A wire substrate of such a material with a preform end in the shape of a regular or concave cone, is first melted with a focused laser beam. Under the influence of a high positive potential, a Taylor cone in a liquid/molten state is formed at that end. The cone is then quenched upon cessation of the laser power, thus freezing the Taylor cone. The tip of the frozen Taylor cone is reheated by the laser to allow its precise localized melting and shaping. Tips thus obtained yield desirable end-forms suitable as electron field emission sources for a variety of applications. In-situ regeneration of the tip is readily accomplished. These tips can also be employed as regenerable bright ion sources using field ionization/desorption of introduced chemical species.

  9. Observations. Changes in Snow, Ice and Frozen Ground. Chapter 4

    Energy Technology Data Exchange (ETDEWEB)

    Lemke, P.; Ren, J.; Alley, R.B.; Allison, I.; Carrasco, J.; Flato, G.; Fujii, Y.; Kaser, G.; Mote, P.; Thomas, R.H.; Zhang, T.

    2007-09-15

    In the climate system, the cryosphere (which consists of snow, river and lake ice, sea ice, glaciers and ice caps, ice shelves and ice sheets, and frozen ground) is intricately linked to the surface energy budget, the water cycle, sea level change and the surface gas exchange. The cryosphere integrates climate variations over a wide range of time scales, making it a natural sensor of climate variability and providing a visible expression of climate change. In the past, the cryosphere has undergone large variations on many time scales associated with ice ages and with shorter-term variations like the Younger Dryas or the Little Ice Age (see Chapter 6). Recent decreases in ice mass are correlated with rising surface air temperatures. This is especially true for the region north of 65N, where temperatures have increased by about twice the global average from 1965 to 2005.

  10. 'Frozen' media subsidies during a time of media change

    DEFF Research Database (Denmark)

    Nielsen, Rasmus Kleis

    2014-01-01

    Media systems around the world have changed in significant ways in the early 21st century. In this article, I analyse how various forms of media subsidies have changed in response to these transformations in a sample of six different affluent democracies. On the basis of interviews, official...... documents and secondary sources, I show that media subsidies have largely remained frozen in their late-20th century form. The absence of major reform means that media subsidies are increasingly subject to policy drift, a process by which the operations and effectiveness of policies change not because...... of deliberate reform, but because of changing conditions on the ground. Analysis of interviews with relevant stakeholders suggests that the main obstacles to reform across all six countries are: (1) limited political attention to the problem; (2) strong incumbent industries protecting their interests; and (3...

  11. Geographical parthenogenesis: General purpose genotypes and frozen niche variation

    DEFF Research Database (Denmark)

    Vrijenhoek, Robert C.; Parker, Dave

    2009-01-01

    Clonally reproducing all-female lineages of plants and animals are often more frequent at higher latitudes and altitudes, on islands, and in disturbed habitats. Attempts to explain this pattern, known as geographical parthenogenesis, generally treat the parthenogens as fugitive species that occupy...... hypotheses concerning the evolution of niche breadth in asexual species - the "general-purpose genotype" (GPG) and "frozen niche-variation" (FNV) models. The two models are often portrayed as mutually exclusive, respectively viewing clonal lineages as generalists versus specialists. Nonetheless......, they are complex syllogisms that share common assumptions regarding the likely origins of clonal diversity and the strength of interclonal selection in shaping the ecological breadth of asexual populations. Both models find support in ecological and phylogeographic studies of a wide range of organisms...

  12. Frozen up dilaton and the GUT/Planck mass ratio

    Science.gov (United States)

    Davidson, Aharon; Ygael, Tomer

    2017-09-01

    By treating modulus and phase on equal footing, as prescribed by Dirac, local scale invariance can consistently accompany any Brans-Dicke ω-theory. We show that in the presence of a soft scale symmetry breaking term, the classical solution, if it exists, cannot be anything else but general relativistic. The dilaton modulus gets frozen up by the Weyl-Proca vector field, thereby constituting a gravitational quasi-Higgs mechanism. Assigning all grand unified scalars as dilatons, they enjoy Weyl universality, and upon symmetry breaking, the Planck (mass)2 becomes the sum of all their individual (VEV)2s. The emerging GUT/Planck (mass)2 ratio is thus ∼ ωgGUT2 / 4 π.

  13. Frozen up dilaton and the GUT/Planck mass ratio

    Directory of Open Access Journals (Sweden)

    Aharon Davidson

    2017-09-01

    Full Text Available By treating modulus and phase on equal footing, as prescribed by Dirac, local scale invariance can consistently accompany any Brans–Dicke ω-theory. We show that in the presence of a soft scale symmetry breaking term, the classical solution, if it exists, cannot be anything else but general relativistic. The dilaton modulus gets frozen up by the Weyl–Proca vector field, thereby constituting a gravitational quasi-Higgs mechanism. Assigning all grand unified scalars as dilatons, they enjoy Weyl universality, and upon symmetry breaking, the Planck (mass2 becomes the sum of all their individual (VEV2s. The emerging GUT/Planck (mass2 ratio is thus ∼ωgGUT2/4π.

  14. Microfluidics for rapid cytokeratin immunohistochemical staining in frozen sections.

    Science.gov (United States)

    Brajkovic, Saska; Dupouy, Diego G; de Leval, Laurence; Gijs, Martin Am

    2017-08-01

    Frozen sections (FS) of tumor samples represent a cornerstone of pathological intraoperative consultation and have an important role in the microscopic analysis of specimens during surgery. So far, immunohistochemical (IHC) stainings on FS have been demonstrated for a few markers using manual methods. Microfluidic technologies have proven to bring substantial improvement in many fields of diagnostics, though only a few microfluidic devices have been designed to improve the performance of IHC assays. In this work, we show optimization of a complete pan-cytokeratin chromogenic immunostaining protocol on FS using a microfluidic tissue processor into a protocol taking <12 min. Our results showed specificity and low levels of background. The dimensions of the microfluidic prototype device are compatible with the space constraints of an intraoperative pathology laboratory. We therefore anticipate that the adoption of microfluidic technologies in the field of surgical pathology can significantly improve the way FSs influence surgical procedures.

  15. Validation of the sperm quality analyzer and the hypo-osmotic swelling test for frozen-thawed ram and minke whale (Balaenoptera bonarensis) spermatozoa.

    Science.gov (United States)

    Fukui, Yutaka; Togawa, Morihiko; Abe, Norihito; Takano, Yuuki; Asada, Masatsugu; Okada, Aki; Iida, Kenji; Ishikawa, Hajime; Ohsumi, Seiji

    2004-02-01

    The object of the present study was to investigate the validation of the sperm quality analyzer (SQA) and the hypo-osmotic swelling (HOS) test with standard sperm analysis methods in frozen-thawed ram and minke whale spermatozoa. In rams, highly significant correlations were observed in the percentage of motile spermatozoa (Psperm concentration (Pwhale spermatozoa, the SVI (sperm viability index) values by the standard method were significantly (Psperm motility index (SMI) values by SQA. The percentage of motile spermatozoa was also significantly correlated (Pwhale spermatozoa, and that the SQA and HOS test values are significantly correlated in ram spermatozoa. However, sperm concentration and morphologically normal spermatozoa are not assessed accurately by SQA in minke whales.

  16. Human evolution in Siberia: from frozen bodies to ancient DNA

    Directory of Open Access Journals (Sweden)

    Bouakaze Caroline

    2010-01-01

    Full Text Available Abstract Background The Yakuts contrast strikingly with other populations from Siberia due to their cattle- and horse-breeding economy as well as their Turkic language. On the basis of ethnological and linguistic criteria as well as population genetic studies, it has been assumed that they originated from South Siberian populations. However, many questions regarding the origins of this intriguing population still need to be clarified (e.g. the precise origin of paternal lineages and the admixture rate with indigenous populations. This study attempts to better understand the origins of the Yakuts by performing genetic analyses on 58 mummified frozen bodies dated from the 15th to the 19th century, excavated from Yakutia (Eastern Siberia. Results High quality data were obtained for the autosomal STRs, Y-chromosomal STRs and SNPs and mtDNA due to exceptional sample preservation. A comparison with the same markers on seven museum specimens excavated 3 to 15 years ago showed significant differences in DNA quantity and quality. Direct access to ancient genetic data from these molecular markers combined with the archaeological evidence, demographical studies and comparisons with 166 contemporary individuals from the same location as the frozen bodies helped us to clarify the microevolution of this intriguing population. Conclusion We were able to trace the origins of the male lineages to a small group of horse-riders from the Cis-Baïkal area. Furthermore, mtDNA data showed that intermarriages between the first settlers with Evenks women led to the establishment of genetic characteristics during the 15th century that are still observed today.

  17. The Fresh Frozen Bone Allograft (FFBA) to reconstruc the atrophic maxilla: evaluation of the clinical and histological viability at six and nine months in the maxillary sinus

    OpenAIRE

    Bassi, Ana Paula Farnezi; Vieira, Rogeria A; Matsumoto, Marisa Akemi; Stecklberg, Iris Monica; Ferreira,Gabriel Ramalho; FAVERANI,Leonardo Perez

    2013-01-01

    This work aimed to analyze clinically and histologically the allogen bone graft behavior at 6 and 9 months. A leukoderm, female, 55 years old patient sought dental care for oral rehabilitation with dental implants and implant supported fixed prosthesis in the maxilla. Bilateral sinus lifting procedure were performed in a same patient, the analysis were made after 6 and 9 months, respectively. At 6 months, there was lack of a better bone remodeling in the region, associated to the presence of ...

  18. Effects of Storage Temperatures on the Quality of Frozen Sardine, Mackerel, and Saury

    Science.gov (United States)

    Kozima, Tsuneo; Ohataka, Tateo

    The three Japanese coastal fish species, sardine (Sardinops melanosticta), mackerel (Scomber japonicus) and saury (Cololabis saira) was frozen under commercial condition and stored at -18, -23, -30 and -40°C for 12 months. During frozen storage the quality was measured by determining the K value (freshness index), peroxide value (POV) of fat, a mount of free drip, water-holding capacity of muscle, weight ratio of cooking loss, histological feature of frozen and thawed muscle, and organoleptic test at regular intervals (each 2 months). Storage life of frozen sardine was 6 months at -18°C and 12 months at below -23°C. On frozen mackerel it was 6 months at -18°C. 8 months at -23°C and 12 months at below -30°C. On saury it was 6 months at 18°C and 12 months at below -23°C.

  19. Muscle Fiber Orientation Angle Dependence of the Tensile Fracture Behavior of Frozen Fish Muscle

    Science.gov (United States)

    Hagura, Yoshio; Okamoto, Kiyoshi; Suzuki, Kanichi; Kubota, Kiyoshi

    We have proposed a new cutting method for frozen fish named "cryo-cutting". This method applied tensile fracture force or bending fracture force to the frozen fish at appropriate low temperatures. In this paper, to clarify cryo-cutting mechanism, we analyzed tensile fracture behavior of the frozen fish muscle. In the analysis, the frozen fish muscle was considered unidirectionally fiber-reinforced composite material which consisted of fiber (muscle fiber) and matrix (connective tissue). Fracture criteria (maximum stress criterion, Tsai-Hill criterion) for the unidirectionally fiber-reinforced composite material were used. The following results were obtained: (1) By using Tsai-Hill criterion, muscle fiber orientation angle dependence of the tensile fracture stress could be calculated. (2) By using the maximum stress theory jointly with Tsai-Hill criterion, muscle fiber orientation angle dependence of the fracture mode of the frozen fish muscle could be estimated.

  20. Accuracy of frozen section diagnosis: a retrospective analysis of 4785 cases

    DEFF Research Database (Denmark)

    Winther, Charlotte; Graem, Niels

    2011-01-01

    During a 1-year period 4785 intraoperative consultations were performed. The pathology reports were retrospectively reviewed to determine the accuracy of frozen section diagnosis in various tissue types. Skin for evaluation of section margins and axillary sentinel lymph nodes for evaluation...... of metastatic disease were most frequently sent for frozen section diagnosis. The number of discordant cases were 182, 178 were false negative and four were false positive. When frozen section diagnoses were compared with permanent section diagnoses, the overall diagnostic concordance was 95.1%. The number...... of deferred specimens was 57. The accuracy of frozen section diagnosis varied between tissue types, and axillary sentinel lymph nodes accounted for the greatest number of discordances. In conclusion, the frozen section diagnosis is a reliable method with varying concordance and deferral rates between tissue...