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Sample records for serum-free freezing medium

  1. Growth in serum-free medium improves isolation of Chlamydia pneumoniae.

    OpenAIRE

    Maass, M; Essig, A; Marre, R; Henkel, W

    1993-01-01

    Infectivity titers were determined for eight Chlamydia pneumoniae strains simultaneously grown in serum-free and serum-supplemented cell culture media. Use of serum-free medium resulted in a 10- to 50-fold increase in the susceptibility of HL cells to chlamydial infection. Comparative primary isolation of a wild-type strain also produced higher inclusion counts in a serum-free environment. Serum-free cultivation is recommended to increase the efficiency of C. pneumoniae isolation from clinica...

  2. Compound serum and hemin free medium for cultivation of ...

    African Journals Online (AJOL)

    Serum free cultivation of Leishmania is cost-effective and improves large scale production of well defined parasite material. Moreover, the production of recombinant pharmaceutical proteins requires cultivation of the host in a culture medium free of animal materials, so several culture media for Leishmania tarentolae ...

  3. Rabbit chondrocytes maintained in serum-free medium. I. Synthesis and secretion of hydrodynamically-small proteoglycans

    International Nuclear Information System (INIS)

    Malemud, C.J.; Papay, R.S.

    1986-01-01

    The biosynthesis of sulfated proteoglycan in vitro by rabbit articular chondrocytes in first passage monolayer culture maintained in fetal bovine serum (FBS) or in serum-free conditions was compared. Neosynthesized proteoglycan in the culture medium in the most dense fraction of an associative CsCl density gradient (fraction dAl) declined with increasing time under serum-free conditions, but not when cells were maintained in the presence of serum. After one day, the major peak of incorporated 35 SO 4 in medium fraction dAl eluted as a retarded peak on Sepharose CL-2B, whether cells were maintained under serum-free or serum-containing conditions. The hydrodynamic size of proteoglycan monomer fraction dAlDl obtained after one day of exposure to serum-free culture media was smaller than dAlDl from serum-containing cultures. The hydrodynamic size of dAlDl obtained from serum-free culture media became even progressively smaller after 2 and 3 days' exposure to these conditions. Hydrodynamically small sulfated proteoglycans were identified in the cell-associated dAlDl fraction as early as one day after switching chondrocytes from serum-containing to serum-free medium. Proteoglycan monomer from serum-containing medium reaggregated more efficiently under both conditions. No change in the size of glycosaminoglycan chains was seen in the smaller proteoglycan subpopulations, nor was there any indication of marked changes in the glycosaminoglycan types

  4. Immunohistochemical expression of stem cell, endothelial cell, and chemosensitivity markers in primary glioma spheroids cultured in serum-containing and serum-free medium

    DEFF Research Database (Denmark)

    Christensen, Karina; Aaberg-Jessen, Charlotte; Andersen, Claus

    2010-01-01

    To investigate the influence of serum-free medium (SFM) supplemented with epidermal growth factor and basic fibroblast growth factor compared with conventional serum-containing medium (SCM) on the phenotype of organotypic primary spheroids from seven gliomas.......To investigate the influence of serum-free medium (SFM) supplemented with epidermal growth factor and basic fibroblast growth factor compared with conventional serum-containing medium (SCM) on the phenotype of organotypic primary spheroids from seven gliomas....

  5. A procedure for culturing rat neocortex explants in a serum-free nutrient medium

    NARCIS (Netherlands)

    Romijn, H. J.; de Jong, B. M.; Ruijter, J. M.

    1988-01-01

    A procedure is described for long-term culturing of rat neocortex explants in a serum-free growth medium. Slices spanning the entire cortical depth from pial to ventricular side are prepared from 6-day-old rat pups. After preincubation in Hanks' balanced salt solution with extra glucose, the

  6. Human dental pulp stem cells cultured in serum-free supplemented medium

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    Virginie eBonnamain

    2013-12-01

    Full Text Available Growing evidence show that human dental pulp stem cells (DPSCs could provide a source of adult stem cells for the treatment of neurodegenerative pathologies. In this study, DPSCs were expanded and cultured with a protocol generally used for the culture of neural stem/progenitor cells.Methodology: DPSC cultures were established from third molars. The pulp tissue was enzymatically digested and cultured in serum-supplemented basal medium for 12 hours. Adherent (ADH and non-adherent (non-ADH cell populations were separated according to their differential adhesion to plastic and then cultured in serum-free defined N2 medium with epidermal growth factor (EGF and basic fibroblast growth factor (bFGF. Both ADH and non-ADH populations were analyzed by FACS and/or PCR.Results: FACS analysis of ADH-DPSCs revealed the expression of the mesenchymal cell marker CD90, the neuronal marker CD56, the transferrin receptor CD71, and the chemokine receptor CXCR3, whereas hematopoietic stem cells markers CD45, CD133 and CD34 were not expressed. ADH-DPSCs expressed transcripts coding for the Nestin gene, whereas expression levels of genes coding for the neuronal markers β-III tubulin and NF-M, and the oligodendrocyte marker PLP-1 were donor dependent. ADH-DPSCs did not express the transcripts for GFAP, an astrocyte marker. Cells of the non-ADH population that grew as spheroids expressed Nestin, β-III tubulin, NF-M and PLP-1 transcripts. DPSCs migrated out of the spheroids exhibited an odontoblast-like morphology and expressed a higher level of DSPP and osteocalcin transcripts than ADH-DPSCs. Conclusion: Collectively, these data indicate that human DPSCs can be expended and cultured in serum-free supplemented medium with EGF and bFGF. ADH-DPSCs and non-ADH populations contained neuronal and/or oligodendrocyte precursors at different stages of commitment and interestingly, cells from spheroid structures seem to be more engaged into the odontoblastic lineage than the

  7. Increasing synthetic serum substitute (SSS) concentrations in P1 glucose/phosphate-free medium improves implantation rate: a comparative study.

    Science.gov (United States)

    Ben-Yosef, D; Yovel, I; Schwartz, T; Azem, F; Lessing, J B; Amit, A

    2001-11-01

    To assess the comparative efficacy of IVF medium (MediCult, with 5.2 mM glucose) and a glucose/phosphate-free medium, P1 (Irvine Scientific), and to investigate the influence of increasing the serum supplementation (synthetic serum substitute; SSS; Irvine Scientific) to P1 on embryo development and implantation. Patients were randomly assigned to IVF medium (Group 1, cycles n = 172) or P1 supplemented with 10% SSS (Group 2, cycles n = 229) according to the medium scheduled for use on the day of oocyte retrieval. Another 555 IVF consequent cycles (Group 3) were performed using increased SSS concentrations (20%) in P1 medium. In this large series of IVF cycles, we herein demonstrate that significantly higher pregnancy and implantation rates were found when embryos were cultured in glucose/phosphate-free medium P1 supplemented with 20% SSS compared to supplementation with the lower SSS concentration and with IVF medium.

  8. The influence of serum substituents on serum-free Vero cell conditioned culture media manufactured from Dulbecco's modified Eagle medium in mouse embryo culture.

    Science.gov (United States)

    Lee, Jong-Seon; Kim, Ju-Hwan; Seo, Young-Seok; Yang, Jung-Bo; Kim, Yong-Il; Kim, Hye-Jin; Lee, Ki-Hwan

    2013-09-01

    This study was conducted to examine the influences of supplementation of the serum substituents and available period of serum-free Vero cell conditioned media (SF-VCM) manufactured from Dulbecco's modified Eagle medium cultured with Vero cells for in vitro development of mouse preimplantation embryos. A total of 1,099 two-cell embryos collected from imprinting control region mice were cultured in SF-VCM with 10% and 20% human follicular fluid (hFF), serum substitute supplement (SSS), and serum protein substitute (SPS). Development of embryos was observed every 24 hours. Results between different groups were analyzed by chi-square test, and considered statistically significant when P-value was less than 0.05. The rates of embryonic development cultured in SF-VCM supplemented with serum substituents were significantly higher compare with serum-free group (P media up to 4 weeks did not affect on embryonic development.

  9. Differentiation of Dental Pulp Stem Cells into Neuron-Like Cells in Serum-Free Medium

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    Shahrul Hisham Zainal Ariffin

    2013-01-01

    Full Text Available Dental pulp tissue contains dental pulp stem cells (DPSCs. Dental pulp cells (also known as dental pulp-derived mesenchymal stem cells are capable of differentiating into multilineage cells including neuron-like cells. The aim of this study was to examine the capability of DPSCs to differentiate into neuron-like cells without using any reagents or growth factors. DPSCs were isolated from teeth extracted from 6- to 8-week-old mice and maintained in complete medium. The cells from the fourth passage were induced to differentiate by culturing in medium without serum or growth factors. RT-PCR molecular analysis showed characteristics of Cd146+, Cd166+, and Cd31− in DPSCs, indicating that these cells are mesenchymal stem cells rather than hematopoietic stem cells. After 5 days of neuronal differentiation, the cells showed neuron-like morphological changes and expressed MAP2 protein. The activation of Nestin was observed at low level prior to differentiation and increased after 5 days of culture in differentiation medium, whereas Tub3 was activated only after 5 days of neuronal differentiation. The proliferation of the differentiated cells decreased in comparison to that of the control cells. Dental pulp stem cells are induced to differentiate into neuron-like cells when cultured in serum- and growth factor-free medium.

  10. Production of inactivated influenza H5N1 vaccines from MDCK cells in serum-free medium.

    Directory of Open Access Journals (Sweden)

    Alan Yung-Chih Hu

    Full Text Available BACKGROUND: Highly pathogenic influenza viruses pose a constant threat which could lead to a global pandemic. Vaccination remains the principal measure to reduce morbidity and mortality from such pandemics. The availability and surging demand for pandemic vaccines needs to be addressed in the preparedness plans. This study presents an improved high-yield manufacturing process for the inactivated influenza H5N1 vaccines using Madin-Darby canine kidney (MDCK cells grown in a serum-free (SF medium microcarrier cell culture system. PRINCIPAL FINDING: The current study has evaluated the performance of cell adaptation switched from serum-containing (SC medium to several commercial SF media. The selected SF medium was further evaluated in various bioreactor culture systems for process scale-up evaluation. No significant difference was found in the cell growth in different sizes of bioreactors studied. In the 7.5 L bioreactor runs, the cell concentration reached to 2.3 × 10(6 cells/mL after 5 days. The maximum virus titers of 1024 Hemagglutinin (HA units/50 µL and 7.1 ± 0.3 × 10(8 pfu/mL were obtained after 3 days infection. The concentration of HA antigen as determined by SRID was found to be 14.1 µg/mL which was higher than those obtained from the SC medium. A mouse immunogenicity study showed that the formalin-inactivated purified SF vaccine candidate formulated with alum adjuvant could induce protective level of virus neutralization titers similar to those obtained from the SC medium. In addition, the H5N1 viruses produced from either SC or SF media showed the same antigenic reactivity with the NIBRG14 standard antisera. CONCLUSIONS: The advantages of this SF cell-based manufacturing process could reduce the animal serum contamination, the cost and lot-to-lot variation of SC medium production. This study provides useful information to manufacturers that are planning to use SF medium for cell-based influenza vaccine production.

  11. Serum-free media formulations are cell line-specific and require optimization for microcarrier culture.

    Science.gov (United States)

    Tan, Kah Yong; Teo, Kim Leng; Lim, Jessica F Y; Chen, Allen K L; Choolani, Mahesh; Reuveny, Shaul; Chan, Jerry; Oh, Steve Kw

    2015-08-01

    Mesenchymal stromal cells (MSCs) are being investigated as potential cell therapies for many different indications. Current methods of production rely on traditional monolayer culture on tissue-culture plastic, usually with the use of serum-supplemented growth media. However, the monolayer culturing system has scale-up limitations and may not meet the projected hundreds of billions to trillions batches of cells needed for therapy. Furthermore, serum-free medium offers several advantages over serum-supplemented medium, which may have supply and contaminant issues, leading to many serum-free medium formulations being developed. We cultured seven MSC lines in six different serum-free media and compared their growth between monolayer and microcarrier culture. We show that (i) expansion levels of MSCs in serum-free monolayer cultures may not correlate with expansion in serum-containing media; (ii) optimal culture conditions (serum-free media for monolayer or microcarrier culture) differ for each cell line; (iii) growth in static microcarrier culture does not correlate with growth in stirred spinner culture; (iv) and that early cell attachment and spreading onto microcarriers does not necessarily predict efficiency of cell expansion in agitated microcarrier culture. Current serum-free media developed for monolayer cultures of MSCs may not support MSC proliferation in microcarrier cultures. Further optimization in medium composition will be required for microcarrier suspension culture for each cell line. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  12. Bovine babesiosis: Cattle protected in the field with a frozen vaccine containing Babesia bovis and Babesia bigemina cultured in vitro with a serum-free medium.

    Science.gov (United States)

    Rojas-Martínez, Carmen; Rodríguez-Vivas, Roger Iván; Millán, Julio Vicente Figueroa; Bautista-Garfias, Carlos Ramón; Castañeda-Arriola, Roberto Omar; Lira-Amaya, José Juan; Urióstegui, Patricia Vargas; Carrasco, Juan José Ojeda; Martínez, Jesús Antonio Álvarez

    2018-04-01

    An attenuated live vaccine containing Babesia bovis and B. bigemina cultured in vitro with a serum-free medium was assessed for its clinical protection conferred of naïve cattle, under natural tick-challenge in a high endemicity zone to Babesia spp. Three groups of six animals were treated as follows: group I (GI) received a vaccine derived from parasites cultured with a free-serum medium; group II (GII) were immunized with the standard vaccine, with parasites cultured in a medium supplemented with 40% (v/v) bovine serum; and a control group (GIII) inoculated with non-infected bovine erythrocytes. Inocula were administered by IM route. Experimental animals were kept during 23days after vaccination in a cattle farm free of ticks and Babesia spp. Thereafter, cattle were moved to a high endemicity farm for natural exposure to Babesia spp. transmitted by Rhipicephalus microplus ticks. Protection against clinical babesiosis was observed in bovines belonging to GI (100%) and GII (83.33%), while the control animals (GIII) were not protected, and showed severe clinical signs, closely related to babesiosis, were observed for at least three consecutive days during the challenge. These were fever, anemia, which were measured simultaneously, and circulating parasites were detected by optic light microscopy. All cattle showed B. bovis and B. bigemina in stained blood films during the challenge; B. bovis antibody titers were higher than those to B. bigemina in GI and GII, and lower titers were determined in GIII. The protective capacity of the vaccine derived from B. bovis and B. bigemina cultured in vitro in a serum-free medium was demonstrated. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Development of a serum-free medium for in vitro expansion of human cytotoxic T lymphocytes using a statistical design

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    Lee Gyun

    2010-09-01

    Full Text Available Abstract Background Serum-containing medium (SCM, which has a number of poorly defined components with varying concentrations, hampers standardization of lymphocyte cultures. In order to develop a serum-free medium (SFM for the expansion of human lymphocytes from peripheral blood mononuclear cells (PBMCs, a statistical optimization approach based on a fractional factorial method and a response surface method was adopted. A basal medium was prepared by supplementing RPMI1640 medium with insulin, albumin, ferric citrate, ethanolamine, fatty acids, glutamine, sodium pyruvate, 2-mercaptoethanol, 1-thioglycerol, nonessential amino acids, and vitamins. We identified additional positive determinants and their optimal concentrations for cell growth through a statistical analysis. Results From a statistical analysis using the fractional factorial method, cholesterol and polyamine supplement were identified as positive determinants for cell growth. Their optimal concentrations were determined by the response surface method. The maximum viable cell concentration in the developed SFM was enhanced by more than 1.5-fold when compared to that in RPMI1640 supplemented with 10% fetal bovine serum (FBS. Furthermore, a cytotoxicity assay and an enzyme-linked immunospot assay revealed that the effector function of cytotoxic T lymphocytes generated from PBMCs grown in SFM, by stimulation of peptide-presenting dendritic cells, was retained or even better than that in SCM. Conclusions The use of a developed SFM with cholesterol and polyamine supplement for human lymphocyte culture resulted in better growth without loss of cellular function when compared to SCM.

  14. Development of a serum-free medium for in vitro expansion of human cytotoxic T lymphocytes using a statistical design.

    Science.gov (United States)

    Jeon, Min Kyoung; Lim, Jong-Baeck; Lee, Gyun Min

    2010-09-21

    Serum-containing medium (SCM), which has a number of poorly defined components with varying concentrations, hampers standardization of lymphocyte cultures. In order to develop a serum-free medium (SFM) for the expansion of human lymphocytes from peripheral blood mononuclear cells (PBMCs), a statistical optimization approach based on a fractional factorial method and a response surface method was adopted. A basal medium was prepared by supplementing RPMI1640 medium with insulin, albumin, ferric citrate, ethanolamine, fatty acids, glutamine, sodium pyruvate, 2-mercaptoethanol, 1-thioglycerol, nonessential amino acids, and vitamins. We identified additional positive determinants and their optimal concentrations for cell growth through a statistical analysis. From a statistical analysis using the fractional factorial method, cholesterol and polyamine supplement were identified as positive determinants for cell growth. Their optimal concentrations were determined by the response surface method. The maximum viable cell concentration in the developed SFM was enhanced by more than 1.5-fold when compared to that in RPMI1640 supplemented with 10% fetal bovine serum (FBS). Furthermore, a cytotoxicity assay and an enzyme-linked immunospot assay revealed that the effector function of cytotoxic T lymphocytes generated from PBMCs grown in SFM, by stimulation of peptide-presenting dendritic cells, was retained or even better than that in SCM. The use of a developed SFM with cholesterol and polyamine supplement for human lymphocyte culture resulted in better growth without loss of cellular function when compared to SCM.

  15. Defined culture medium for stem cell differentiation: applicability of serum-free conditions in the mouse embryonic stem cell test.

    Science.gov (United States)

    Riebeling, Christian; Schlechter, Katharina; Buesen, Roland; Spielmann, Horst; Luch, Andreas; Seiler, Andrea

    2011-06-01

    The embryonic stem cell test (EST) is a validated method to assess the developmental toxicity potency of chemicals. It was developed to reduce animal use and allow faster testing for hazard assessment. The cells used in this method are maintained and differentiated in media containing foetal calf serum. This animal product is of considerable variation in quality, and individual batches require extensive testing for their applicability in the EST. Moreover, its production involves a large number of foetuses and possible animal suffering. We demonstrate the serum-free medium and feeder cell-free maintenance of the mouse embryonic stem cell line D3 and investigate the use of specific growth factors for induction of cardiac differentiation. Using a combination of bone morphogenetic protein-2, bone morphogenetic protein-4, activin A and ascorbic acid, embryoid bodies efficiently differentiated into contracting myocardium. Additionally, examining levels of intracellular marker proteins by flow cytometry not only confirmed differentiation into cardiomyocytes, but demonstrated significant differentiation into neuronal cells in the same time frame. Thus, this approach might allow for simultaneous detection of developmental effects on both early mesodermal and neuroectodermal differentiation. The serum-free conditions for maintenance and differentiation of D3 cells described here enhance the transferability and standardisation and hence the performance of the EST. Copyright © 2011 Elsevier Ltd. All rights reserved.

  16. Heat and Mass Transfer Model in Freeze-Dried Medium

    Science.gov (United States)

    Alfat, Sayahdin; Purqon, Acep

    2017-07-01

    There are big problems in agriculture sector every year. One of the major problems is abundance of agricultural product during the peak of harvest season that is not matched by an increase in demand of agricultural product by consumers, this causes a wasted agricultural products. Alternative way was food preservation by freeze dried method. This method was already using heat transfer through conduction and convection to reduce water quality in the food. The main objective of this research was to design a model heat and mass transfer in freeze-dried medium. We had two steps in this research, the first step was design of medium as the heat injection site and the second was simulate heat and mass transfer of the product. During simulation process, we use physical property of some agriculture product. The result will show how temperature and moisture distribution every second. The method of research use finite element method (FEM) and will be illustrated in three dimensional.

  17. Effect of haemolysis and repeated freeze-thawing cycles on wild boar serum antibody testing by ELISA

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    Boadella Mariana

    2011-11-01

    Full Text Available Abstract Background Monitoring wildlife diseases is needed to identify changes in disease occurrence. Wildlife blood samples are valuable for this purpose but are often gathered haemolysed. To maximise information, sera often go through repeated analysis and freeze-thaw cycles. Herein, we used samples of clean and haemolysed Eurasian wild boar (Sus scrofa serum stored at -20°C and thawed up to five times to study the effects of both treatments on the outcome of a commercial ELISA test for the detection of antibodies against Suid Herpesvirus 1 (ADV. Results The estimated prevalence of antibodies against ADV was 50-53% for clean and haemolysed sera. Hence, haemolysis did not reduce the mean observed serum antibody prevalence. However, 10 samples changed their classification after repeated freeze-thawing. This included 3 (15% of the clean sera and 7 (41% of the haemolysed sera. Conclusions We recommend (1 establishing more restrictive cut-off values when testing wildlife sera, (2 recording serum quality prior to sample banking, (3 recording the number of freezing-thawing cycles and (4 store sera in various aliquots to reduce repeated usage. For instance, sera with more than 3 freeze-thaw cycles and a haemolysis of over 3 on a scale of 4 should better be discarded for serum antibody monitoring. Even clean (almost not haemolysed sera should not go through more than 5 freeze-thaw cycles.

  18. Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.

    Science.gov (United States)

    Kaneko, Ai; Sankai, Yoshiyuki

    2014-01-01

    The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months) primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4) cells/mL (8.9×10(3) cells/cm2) without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm), greater cell viability (≥30%) for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks).

  19. Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.

    Directory of Open Access Journals (Sweden)

    Ai Kaneko

    Full Text Available The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4 cells/mL (8.9×10(3 cells/cm2 without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm, greater cell viability (≥30% for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks.

  20. Direct comparative analysis of conventional and directional freezing for the cryopreservation of whole ovaries

    NARCIS (Netherlands)

    Maffei, S.; Hanenberg, M.; Pennarossa, G.; Roberto V. Silva, J.; Tiziana, A.; Brevini, L.; Pharm, D.; Arav, A.; Gandolfi, F.

    2013-01-01

    INTERVENTION(S): Eighty-one ovaries were randomly assigned to fresh control, conventional freezing (CF), and directional freezing (DF) group. Ovaries of CF and DF groups were perfused via the ovarian artery with Leibovitz L-15 medium, 10% fetal bovine serum, and 1.5 M dimethyl sulfoxide for 5

  1. In vitro culture of human osteosarcoma cell lines: a comparison of functional characteristics for cell lines cultured in medium without and with fetal calf serum.

    Science.gov (United States)

    Bruserud, Oystein; Tronstad, Karl Johan; Berge, Rolf

    2005-06-01

    Experimental in vitro models including well-characterised cell lines can be used to identify possible new therapeutic targets for the treatment of osteosarcoma. Culture media including inactivated serum is often recommended for in vitro culture of osteosarcoma cells, but the serum component then represents a nonstandardised parameter including a wide range of unidentified mediators. To improve the standardisation we have investigated whether serum-free culture media can be used in experimental in vitro studies of osteosarcoma cell lines. The seven osteosarcoma cell lines Cal72, SJSA-1, Saos-2, SK-ES-1, U2OS, 143.98.2, and KHOS-32IH were cultured in vitro in various serum-free media and media supplemented with 10% heat-inactivated fetal calf serum (FCS). Although proliferation often was relatively low in serum-free media (X-vivo 10, X-vivo 15, X-vivo 20, Stem Span SFEM), some cell lines (Cal72, KHOS-32IH, Saos-2) showed proliferation comparable with the recommended FCS-containing media even when using serum-free conditions. The optimal serum-free medium then varied between cell lines. We also compared 6 different FCS-containing media (including Stem Span with 10% FCS) and the optimal FCS-containing medium varied between cell lines. However, all cell lines proliferated well in Stem Span with FCS, and this medium was regarded as optimal for four of the lines. FCS could not be replaced by fatty acids or low density lipoprotein when testing the Stem Span medium. The release of a wide range of soluble mediators showed only minor differences when using serum-free and FCS-containing media (including Stem Span with and without FCS), and serum-free Stem Span could also be used for in vitro studies of mitogen-stimulated T cell activation in the presence of accessory osteosarcoma cells. The use of Stem Span with 10% FCS allowed the release of a wide range of chemokines by osteosarcoma cell lines (Cal72, SJSA-1), and the chemokine release profile was very similar to the

  2. Development of serum-free media for lepidopteran insect cell lines.

    Science.gov (United States)

    Agathos, Spiros N

    2007-01-01

    Lepidopteran insect cell culture technology has progressed to the point of becoming an essential part of one of the most successful eukaryotic expression systems and is increasingly used industrially on a large scale. Therefore, there is a constant need for convenient and low-cost culture media capable of supporting good insect cell growth and ensuring high yield of baculovirus as well as the strong expression of recombinant proteins. Vertebrate sera or invertebrate hemolymph were essential supplements in first-generation insect cell media. These supplements, however, are cumbersome and expensive for routine large-scale culture; thus, their use is now circumvented by substituting the essential growth factors present in these supplements with serum-free substances. Such non-serum supplements are typically of non-animal origin and include protein hydrolysates, lipid emulsions, and specialized substances (e.g., surfactants and shear damage protecting chemicals). These supplements need to complement the defined, synthetic basal medium to ensure that the fundamental nutritional needs of the cells are satisfied. Although there is a significant number of proprietary serum-free and low-protein or protein-free media on the market, the lack of information concerning their detailed composition is a drawback in their adoption for different applications, including their adaptation to the metabolic and kinetic analysis and monitoring of a given insect cell based bioprocess. Hence, there is wide appeal for formulating serum-free media based on a rational assessment of the metabolic requirements of the lepidopteran cells during both the growth and the production phases. Techniques such as statistical experimental design and genetic algorithms adapted to the cellular behavior and the bioreactor operation mode (batch, fed-batch, or perfusion) permit the formulation of versatile serum- and protein-free media. These techniques are illustrated with recent developments of serum-free

  3. Culture in embryonic kidney serum and xeno-free media as renal cell carcinoma and renal cell carcinoma cancer stem cells research model.

    Science.gov (United States)

    Krawczyk, Krzysztof M; Matak, Damian; Szymanski, Lukasz; Szczylik, Cezary; Porta, Camillo; Czarnecka, Anna M

    2018-04-01

    The use of fetal bovine serum hinders obtaining reproducible experimental results and should also be removed in hormone and growth factor studies. In particular hormones found in FBS act globally on cancer cell physiology and influence transcriptome and metabolome. The aim of our study was to develop a renal carcinoma serum free culture model optimized for (embryonal) renal cells in order to select the best study model for downstream auto-, para- or endocrine research. Secondary aim was to verify renal carcinoma stem cell culture for this application. In the study, we have cultured renal cell carcinoma primary tumour cell line (786-0) as well as human kidney cancer stem cells in standard 2D monolayer cultures in Roswell Park Memorial Institute Medium or Dulbecco's Modified Eagle's Medium and Complete Human Kidney Cancer Stem Cell Medium, respectively. Serum-free, animal-component free Human Embryonic Kidney 293 media were tested. Our results revealed that xeno-free embryonal renal cells optimized culture media provide a useful tool in RCC cancer biology research and at the same time enable effective growth of RCC. We propose bio-mimic RCC cell culture model with specific serum-free and xeno-free medium that promote RCC cell viability.

  4. Optimization of protectant, salinity and freezing condition for freeze-drying preservation of Edwardsiella tarda

    Science.gov (United States)

    Yu, Yongxiang; Zhang, Zheng; Wang, Yingeng; Liao, Meijie; Li, Bin; Xue, Liangyi

    2017-10-01

    Novel preservation condition without ultra-low temperature is needed for the study of pathogen in marine fishes. Freeze-drying is such a method usually used for preservation of terrigenous bacteria. However, studies using freeze-drying method to preserving marine microorganisms remain very limited. In this study, we optimized the composition of protectants during the freeze-drying of Edwardsiella tarda, a fish pathogen that causes systemic infection in marine fishes. We found that the optimal composition of protectant mixture contained trehalose (8.0%), skim milk (12.0%), sodium citrate (2.0%), serum (12.0%) and PVP (2.0%). Orthogonal and interaction analyses demonstrated the interaction between serum and skim milk or sodium citrate. The highest survival rate of E. tarda was observed when the concentration of NaCl was 10.0, 30.0 and between 5.0 and 10.0 g L-1 for preparing TSB medium, E. tarda suspension and protectant mixture, respectively. When E. tarda was frozen at -80°C or -40°C for 6 h, its survival rate was higher than that under other tested conditions. Under the optimized conditions, when the protectant mixture was used during freeze-drying process, the survival rate (79.63%-82.30%) of E. tarda was significantly higher than that obtained using single protectant. Scanning electron microscopy (SEM) image indicated that E. tarda was embedded in thick matrix with detectable aggregation. In sum, the protectant mixture may be used as a novel cryoprotective additive for E. tarda.

  5. Generation of hematopoietic stem cells from human embryonic stem cells using a defined, stepwise, serum-free, and serum replacement-free monolayer culture method.

    Science.gov (United States)

    Kim, So-Jung; Jung, Ji-Won; Ha, Hye-Yeong; Koo, Soo Kyung; Kim, Eung-Gook; Kim, Jung-Hyun

    2017-03-01

    Embryonic stem cells (ESCs) can be expanded infinitely in vitro and have the potential to differentiate into hematopoietic stem cells (HSCs); thus, they are considered a useful source of cells for HSC production. Although several technical in vitro methods for engineering HSCs from pluripotent stem cells have been developed, clinical application of HSCs engineered from pluripotent stem cells is restricted because of the possibility of xenogeneic contamination resulting from the use of murine materials. Human ESCs (CHA-hES15) were cultured on growth factor-reduced Matrigel-coated dishes in the mTeSR1 serum-free medium. When the cells were 70% confluent, we initiated HSC differentiation by three methods involving (1) knockout serum replacement (KSR), cytokines, TGFb1, EPO, and FLT3L; (2) KSR, cytokines, and bFGF; or (3) cytokines and bFGF. Among the three differentiation methods, the minimal number of cytokines without KSR resulted in the greatest production of HSCs. The optimized method resulted in a higher proportion of CD34 + CD43 + hematopoietic progenitor cells (HPCs) and CD34 + CD45 + HPCs compared to the other methods. In addition, the HSCs showed the potential to differentiate into multiple lineages of hematopoietic cells in vitro . In this study, we optimized a two-step, serum-free, animal protein-free, KSR-free, feeder-free, chemically defined monolayer culture method for generation of HSCs and hematopoietic stem and progenitor cells (HSPCs) from human ESCs.

  6. Bone Marrow Mesenchymal Stem Cells Enhance the Differentiation of Human Switched Memory B Lymphocytes into Plasma Cells in Serum-Free Medium

    Directory of Open Access Journals (Sweden)

    Guillaume Bonnaure

    2016-01-01

    Full Text Available The differentiation of human B lymphocytes into plasma cells is one of the most stirring questions with regard to adaptive immunity. However, the terminal differentiation and survival of plasma cells are still topics with much to be discovered, especially when targeting switched memory B lymphocytes. Plasma cells can migrate to the bone marrow in response to a CXCL12 gradient and survive for several years while secreting antibodies. In this study, we aimed to get closer to niches favoring plasma cell survival. We tested low oxygen concentrations and coculture with mesenchymal stem cells (MSC from human bone marrow. Besides, all cultures were performed using an animal protein-free medium. Overall, our model enables the generation of high proportions of CD38+CD138+CD31+ plasma cells (≥50% when CD40-activated switched memory B lymphocytes were cultured in direct contact with mesenchymal stem cells. In these cultures, the secretion of CXCL12 and TGF-β, usually found in the bone marrow, was linked to the presence of MSC. The level of oxygen appeared less impactful than the contact with MSC. This study shows for the first time that expanded switched memory B lymphocytes can be differentiated into plasma cells using exclusively a serum-free medium.

  7. Generation of megakaryocytic progenitors from human embryonic stem cells in a feeder- and serum-free medium.

    Directory of Open Access Journals (Sweden)

    Marjorie Pick

    Full Text Available BACKGROUND: The production of human platelets from embryonic stem cells in a defined culture system is a prerequisite for the generation of platelets for therapeutic use. As an important step towards this goal, we report the differentiation of human embryonic stem cells (hESCs towards the megakaryocyte (Mk lineage using a 'spin embryoid body' method in serum-free differentiation medium. METHODOLOGY AND PRINCIPAL FINDINGS: Immunophenotypic analyses of differentiating hESC identified a subpopulation of cells expressing high levels of CD41a that expressed other markers associated with the Mk lineage, including CD110, CD42b and CD61. Differentiated cells were sorted on the basis of their expression of CD41a, CD34 and CD45 and assessed for Mk colony formation, expression of myeloid and Mk genes and ability to endoreplicate DNA. In a collagen-based colony assay, the CD41a⁺ cells sorted from these differentiation cultures produced 100-800 Mk progenitors at day 13 and 25-160 Mk progenitors at day 20 of differentiation per 100,000 cells assayed. Differentiated Mk cells produced platelet-like particles which expressed CD42b and were activated by ADP, similar to platelets generated from precursors in cord blood. These studies were complemented by real time PCR analyses showing that subsets of cells enriched for CD41a⁺ Mk precursors expressed high levels of Mk associated genes such as PF4 and MPL. Conversely, high levels of myeloid and erythroid related transcripts, such as GATA1, TAL1/SCL and PU.1, were detected in sorted fractions containing CD34⁺ and CD45⁺ cells. CONCLUSIONS: We describe a serum- and feeder-free culture system that enabled the generation of Mk progenitors from human embryonic stem cells. These cells formed colonies that included differentiated Mks that fragmented to form platelet-like particles. This protocol represents an important step towards the generation of human platelets for therapeutic use.

  8. Taurine Biosynthesis in a Fish Liver Cell Line (ZFL) Adapted to a Serum-Free Medium.

    Science.gov (United States)

    Liu, Chieh-Lun; Watson, Aaron M; Place, Allen R; Jagus, Rosemary

    2017-05-25

    Although taurine has been shown to play multiple important physiological roles in teleosts, little is known about the molecular mechanisms underlying dietary requirements. Cell lines can provide useful tools for deciphering biosynthetic pathways and their regulation. However, culture media and sera contain variable taurine levels. To provide a useful cell line for the investigation of taurine homeostasis, an adult zebrafish liver cell line (ZFL) has been adapted to a taurine-free medium by gradual accommodation to a commercially available synthetic medium, UltraMEM™-ITES. Here we show that ZFL cells are able to synthesize taurine and be maintained in medium without taurine. This has allowed for the investigation of the effects of taurine supplementation on cell growth, cellular amino acid pools, as well as the expression of the taurine biosynthetic pathway and taurine transporter genes in a defined fish cell type. After taurine supplementation, cellular taurine levels increase but hypotaurine levels stay constant, suggesting little suppression of taurine biosynthesis. Cellular methionine levels do not change after taurine addition, consistent with maintenance of taurine biosynthesis. The addition of taurine to cells grown in taurine-free medium has little effect on transcript levels of the biosynthetic pathway genes for cysteine dioxygenase (CDO), cysteine sulfinate decarboxylase (CSAD), or cysteamine dioxygenase (ADO). In contrast, supplementation with taurine causes a 30% reduction in transcript levels of the taurine transporter, TauT. This experimental approach can be tailored for the development of cell lines from aquaculture species for the elucidation of their taurine biosynthetic capacity.

  9. In-medium Modifications of Hadron Masses and Chemical Freeze-out in Ultra-relativistic Heavy-ion Collisions

    International Nuclear Information System (INIS)

    Florkowski, W.; Broniowski, W.

    1999-10-01

    We confront the hypothesis of chemical freeze-out in ultra-relativistic heavy-ion collisions with the hypothesis of large modifications of hadron masses in nuclear medium. We find that the thermal-model predictions for the ratios of particle multiplicities are sensitive to the values of in-medium hadronic masses. In particular, the π + /p ratio decreases by 35% when the masses of all hadrons (except for pseudo-Goldstone bosons) are scaled down by 30%. (author)

  10. Optimization of a serum-free culture medium for mouse embryonic stem cells using design of experiments (DoE) methodology.

    Science.gov (United States)

    Knöspel, Fanny; Schindler, Rudolf K; Lübberstedt, Marc; Petzolt, Stephanie; Gerlach, Jörg C; Zeilinger, Katrin

    2010-12-01

    The in vitro culture behaviour of embryonic stem cells (ESC) is strongly influenced by the culture conditions. Current culture media for expansion of ESC contain some undefined substances. Considering potential clinical translation work with such cells, the use of defined media is desirable. We have used Design of Experiments (DoE) methods to investigate the composition of a serum-free chemically defined culture medium for expansion of mouse embryonic stem cells (mESC). Factor screening analysis according to Plackett-Burman revealed that insulin and leukaemia inhibitory factor (LIF) had a significant positive influence on the proliferation activity of the cells, while zinc and L: -cysteine reduced the cell growth. Further analysis using minimum run resolution IV (MinRes IV) design indicates that following factor adjustment LIF becomes the main factor for the survival and proliferation of mESC. In conclusion, DoE screening assays are applicable to develop and to refine culture media for stem cells and could also be employed to optimize culture media for human embryonic stem cells (hESC).

  11. The influence of serum-free culture conditions on skeletal muscle differentiation in a tissue-engineered model

    NARCIS (Netherlands)

    Gawlitta, D.; Boonen, K.J.M.; Oomens, C.W.J.; Baaijens, F.P.T.; Bouten, C.V.C.

    2008-01-01

    The influence of differentiation medium (DM) components on C2C12 murine myoblast differentiation has only been studied in monolayer cultures. Serum-free formulations have been applied that omit the use of sera with unknown composition. The goal of the present study was to compare the influence of

  12. Stability of buffer-free freeze-dried formulations: A feasibility study of a monoclonal antibody at high protein concentrations.

    Science.gov (United States)

    Garidel, Patrick; Pevestorf, Benjamin; Bahrenburg, Sven

    2015-11-01

    We studied the stability of freeze-dried therapeutic protein formulations over a range of initial concentrations (from 40 to 160 mg/mL) and employed a variety of formulation strategies (including buffer-free freeze dried formulations, or BF-FDF). Highly concentrated, buffer-free liquid formulations of therapeutic monoclonal antibodies (mAbs) have been shown to be a viable alternative to conventionally buffered preparations. We considered whether it is feasible to use the buffer-free strategy in freeze-dried formulations, as an answer to some of the known drawbacks of conventional buffers. We therefore conducted an accelerated stability study (24 weeks at 40 °C) to assess the feasibility of stabilizing freeze-dried formulations without "classical" buffer components. Factors monitored included pH stability, protein integrity, and protein aggregation. Because the protein solutions are inherently self-buffering, and the system's buffer capacity scales with protein concentration, we included highly concentrated buffer-free freeze-dried formulations in the study. The tested formulations ranged from "fully formulated" (containing both conventional buffer and disaccharide stabilizers) to "buffer-free" (including formulations with only disaccharide lyoprotectant stabilizers) to "excipient-free" (with neither added buffers nor stabilizers). We evaluated the impacts of varying concentrations, buffering schemes, pHs, and lyoprotectant additives. At the end of 24 weeks, no change in pH was observed in any of the buffer-free formulations. Unbuffered formulations were found to have shorter reconstitution times and lower opalescence than buffered formulations. Protein stability was assessed by visual inspection, sub-visible particle analysis, protein monomer content, charge variants analysis, and hydrophobic interaction chromatography. All of these measures found the stability of buffer-free formulations that included a disaccharide stabilizer comparable to buffer

  13. Influence of serum extraction from the culture medium and of sublethal X-ray irradiation upon microvilli and invaginations of the membrane of Ehrlich ascites tumor cells in monolayer culture

    International Nuclear Information System (INIS)

    Laudenbach, G.; Pfab, R.; Hess, F.; Schachtschabel, D.O.

    1984-01-01

    In order to find out modifications of microvilli and invaginations, the cellular surfaces of Ehrlich ascites tumor cells in monolayer culture (basal medium of Eagle + 10% fetal calf serum) were investigated with the aid of electron-microscopic cross-sections. The tumor cells had been cultured without serum 24 hours prior to investigation or irradiated with 2 Gy. Morphometric evaluation after cell culture in a serum-free medium showed a reduced number of microvilli and a diminution of sections of microvilli. As already described before, a reduction of cell proliferation, of the microtubule-microfilament system, and of the endocytosis activity occurs under these serum-free conditions. The number of invaginations (related to a constant membrane part) was reduced by nearly 50% after serum extraction. Similarly to serum extraction, sublethal X-ray irradiation reduced the sections of microvilli, whereas the number of microvilli increased slightly. Contrary to the effect of serum extraction, the irradiated cells showed twice as many invaginations as the non-irradiated control cells. These differences in the surface structures are interpreted as a result of modified growth stimulations (+- serum) and radiogenic reparation processes. (orig.) [de

  14. Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1-6

    DEFF Research Database (Denmark)

    Mathiesen, Christian K; Jensen, Tanja B; Prentoe, Jannick

    2014-01-01

    Recently, cell culture systems producing hepatitis C virus particles (HCVcc) were developed. Establishment of serum-free culture conditions is expected to facilitate development of a whole-virus inactivated HCV vaccine. We describe generation of genotype 1-6 serum-free HCVcc (sf-HCVcc) from Huh7.......5 hepatoma cells cultured in adenovirus expression medium. Compared to HCVcc, sf-HCVcc showed 0.6-2.1 log10 higher infectivity titers (4.7-6.2 log10 Focus Forming Units/mL), possibly due to increased release and specific infectivity of sf-HCVcc. In contrast to HCVcc, sf-HCVcc had a homogeneous single......-peak density profile. Entry of sf-HCVcc depended on HCV co-receptors CD81, LDLr, and SR-BI, and clathrin-mediated endocytosis. HCVcc and sf-HCVcc were neutralized similarly by chronic-phase patient sera and by human monoclonal antibodies targeting conformational epitopes. Thus, we developed serum-free culture...

  15. Newly-derived neuroblastoma cell lines propagated in serum-free media recapitulate the genotype and phenotype of primary neuroblastoma tumours

    NARCIS (Netherlands)

    Bate-Eya, Laurel T.; Ebus, Marli E.; Koster, Jan; den Hartog, Ilona J. M.; Zwijnenburg, Danny A.; Schild, Linda; van der Ploeg, Ida; Dolman, M. Emmy M.; Caron, Huib N.; Versteeg, Rogier; Molenaar, Jan J.

    2014-01-01

    Recently protocols have been devised for the culturing of cell lines from fresh tumours under serum-free conditions in defined neural stem cell medium. These cells, frequently called tumour initiating cells (TICs) closely retained characteristics of the tumours of origin. We report the isolation of

  16. Freeze dried blood and development of an artificial diet for blood feeding arthropods

    International Nuclear Information System (INIS)

    DeLoach, J.R.; Spates, G.E.; Kapatsa, G.M.; Sheffield, C.L.; Kabayo, J.P.

    1990-01-01

    The goals of the research were to determine the biochemical differences between freeze dried bovine and porcine blood relative to their nutritional value to Glossina palpalis palpalis and Stomoxys calcitrans and to develop an artificial diet for mass rearing these flies. Freeze dried bovine and porcine blood were found to differ in their amino acid content; total dietary lipids did not significantly differ, but some notable exceptions were found in fatty acid content. Both sonication and addition of foetal bovine serum to freeze dried bovine blood improved its nutritional value for G. p. palpalis. A two component, semi-defined artificial diet was developed for G. p. palpalis and S. calcitrans. The College Station diet consisted of lipid contaminated bovine haemoglobin (BHb) and bovine serum albumin (BSA). To conduct dietary deletion tests, a process was developed for preparing large quantities of ultrapure lipid free bovine haemoglobin. S. calcitrans fed on lipid free BHb plus BSA had zero fecundity. Lipids were re-added to the protein diet in three forms: (1) lipid contaminated BHb, (2) pure erythrocyte ghosts, and (3) pure lipids. It was found that membrane lipid from the erythrocyte is required by S. calcitrans. A defined artificial diet consisting of lipid free BHb, BSA, sphingomyelin, phosphatidyl ethanolamine, phosphatidyl serine and cholesterol gave normal adult survival, as well as near normal fecundity and percentage egg hatch for S. calcitrans. Knowing the identity of the lipids, it is now possible to prepare dietary formulations to alleviate dependency on the blood proteins BHb and BSA. (author). 34 refs, 1 fig., 15 tabs

  17. Keratinocytes propagated in serum-free, feeder-free culture conditions fail to form stratified epidermis in a reconstituted skin model.

    Directory of Open Access Journals (Sweden)

    Rebecca Lamb

    Full Text Available Primary human epidermal stem cells isolated from skin tissues and subsequently expanded in tissue culture are used for human therapeutic use to reconstitute skin on patients and to generate artificial skin in culture for academic and commercial research. Classically, epidermal cells, known as keratinocytes, required fibroblast feeder support and serum-containing media for serial propagation. In alignment with global efforts to remove potential animal contaminants, many serum-free, feeder-free culture methods have been developed that support derivation and growth of these cells in 2-dimensional culture. Here we show that keratinocytes grown continually in serum-free and feeder-free conditions were unable to form into a stratified, mature epidermis in a skin equivalent model. This is not due to loss of cell potential as keratinocytes propagated in serum-free, feeder-free conditions retain their ability to form stratified epidermis when re-introduced to classic serum-containing media. Extracellular calcium supplementation failed to improve epidermis development. In contrast, the addition of serum to commercial, growth media developed for serum-free expansion of keratinocytes facilitated 3-dimensional stratification in our skin equivalent model. Moreover, the addition of heat-inactivated serum improved the epidermis structure and thickness, suggesting that serum contains factors that both aid and inhibit stratification.

  18. Effect of IL-6 on proliferation and IG production of human EBV-transformed cell lines in serum free culture media

    NARCIS (Netherlands)

    Jochems, G. J.; Jordens, R.; van Lier, R. A.; Zeijlemaker, W. P.

    1990-01-01

    To optimalize growth and Ig production of EBV transformed B cells for large scale tissue culture, we analyzed five stable monoclonal EBV-B cell lines for their responsiveness to interleukin (IL)-6 in standard medium with 5% FCS and in several serum-free media. As we previously demonstrated these

  19. Cryopreservation of human sperm: efficacy and use of a new nitrogen-free controlled rate freezer versus liquid nitrogen vapour freezing.

    Science.gov (United States)

    Creemers, E; Nijs, M; Vanheusden, E; Ombelet, W

    2011-12-01

    Preservation of spermatozoa is an important aspect of assisted reproductive medicine. The aim of this study was to investigate the efficacy and use of a recently developed liquid nitrogen and cryogen-free controlled rate freezer and this compared with the classical liquid nitrogen vapour freezing method for the cryopreservation of human spermatozoa. Ten patients entering the IVF programme donated semen samples for the study. Samples were analysed according to the World Health Organization guidelines. No significant difference in total sperm motility after freeze-thawing between the new technique and classical technique was demonstrated. The advantage of the new freezing technique is that it uses no liquid nitrogen during the freezing process, hence being safer to use and clean room compatible. Investment costs are higher for the apparatus but running costs are only 1% in comparison with classical liquid nitrogen freezing. In conclusion, post-thaw motility of samples frozen with the classical liquid nitrogen vapour technique was comparable with samples frozen with the new nitrogen-free freezing technique. This latter technique can thus be a very useful asset to the sperm cryopreservation laboratory. © 2011 Blackwell Verlag GmbH.

  20. Well-plate freeze-drying

    DEFF Research Database (Denmark)

    Trnka, Hjalte; Rantanen, Jukka; Grohganz, Holger

    2015-01-01

    Abstract Context: Freeze-drying in presence of excipients is a common practice to stabilize biomacromolecular formulations. The composition of this formulation is known to affect the quality of the final product. Objective: The aim of this study was to evaluate freeze-drying in well-plates as a h......Abstract Context: Freeze-drying in presence of excipients is a common practice to stabilize biomacromolecular formulations. The composition of this formulation is known to affect the quality of the final product. Objective: The aim of this study was to evaluate freeze-drying in well......-plates as a high throughput platform for formulation screening of freeze-dried products. Methods: Model formulations consisting of mannitol, sucrose and bovine serum albumin were freeze-dried in brass well plates, plastic well plates and vials. Physical properties investigated were solid form, residual moisture......, cake collapse and reconstitution time. Results: Samples freeze-dried in well-plates had an acceptable visual cake appearance. Solid form analysis by high throughput X-ray powder diffraction indicated comparable polymorphic outcome independent of the container. The expected increase in moisture level...

  1. Serum triiodothyronine and free triiodothyronine index in normal pregnancy

    International Nuclear Information System (INIS)

    Karunanidhi, A.; Charles, S.X.; Kanagasabapathy, A.S.

    1981-01-01

    The thyroid function at various stages of pregnancy and after delivery was assessed by measuring serum total triiodothyronine (T 3 ), T 3 resin uptake (T 3 RU) and free triiodothyronine index (FT 3 I). In first, second and third trimesters of pregnancy, the mean values of serum total T 3 were significantly elevated (P 3 I remained unaltered. During labour, both serum total T 3 and FT 3 I observed during labour and after delivery may be due to the changes in blood oestrogen levels. This study indicates that serum free T 3 concentrations in pregnancy remained unaltered in the presence of elevated serum total T 3 levels. FT 3 I determination would thus appear to be a reliable in vitro thyroid function test for the assessment of thyroid function throughout pregnancy. Serum total T 3 was measured by radioimmunoassay using radioiodinated T 3 ( 125 I-T 3 ). (author)

  2. Apoptosis in chondrogenesis of human mesenchymal stem cells: effect of serum and medium supplements.

    Science.gov (United States)

    Wang, Chien-Yuan; Chen, Ling-Lan; Kuo, Pei-Yin; Chang, Jia-Ling; Wang, Yng-Jiin; Hung, Shih-Chieh

    2010-04-01

    Apoptosis is an inevitable process during development and is evident in the formation of articular cartilage and endochondral ossification of growth plate. Mesenchymal stem cells (MSCs) can serve as alternative sources for cell therapy in focal chondral lesions or diffuse osteoarthritis. But there are few, if any, studies investigating apoptosis during chondrogenesis by MSCs. The aim of this study was to find the better condition to prevent apoptosis during chondrogenesis by MSCs. Apoptosis were evaluated in MSCs induced in different chondrogenic media by the use of Annexin V, TUNEL staining, lysosomal labeling with lysotracker and immunostaining of apoptotic markers. We found apparent apoptosis was demonstrated by Annexin V, TUNEL staining and lysosomal labeling during chondrogenesis. Meanwhile, the degree of apoptosis was related to the reagents of the defined chondrogenic medium. Adding serum in medium increased apoptosis, however, TGF-beta1 inhibited apoptosis. The apoptosis was associated with the activation of caspase-3, the increase in the Bax/Bcl-2 ratio, the loss of lysosomal integrity, and the increase of PARP-cleavage. Pro-inflammatory cytokines, IL-1alpha, IL-1beta and TNFalpha did not induce any increase in apoptosis. Interestingly, the inhibition of apoptosis by serum free medium supplemented with ITS was also associated with an increase in the expression of type II collagen, and a decrease in the expression of type X collagen, Runx2, and other osteogenic genes, while TGF-beta1 increased the expression of Sox9, type II and type X collagen and decreased the expression of osteogenic genes. These data suggest apoptosis occurs during chondrogenesis by MSCs by cell death intrinsic pathway activation and this process may be modulated by culture conditions.

  3. Evaluation of human platelet lysate versus fetal bovine serum for culture of mesenchymal stromal cells.

    Science.gov (United States)

    Hemeda, Hatim; Giebel, Bernd; Wagner, Wolfgang

    2014-02-01

    Culture media for therapeutic cell preparations-such as mesenchymal stromal cells (MSCs)-usually comprise serum additives. Traditionally, fetal bovine serum is supplemented in basic research and in most clinical trials. Within the past years, many laboratories adapted their culture conditions to human platelet lysate (hPL), which further stimulates proliferation and expansion of MSCs. Particularly with regard to clinical application, human alternatives for fetal bovine serum are clearly to be preferred. hPL is generated from human platelet units by disruption of the platelet membrane, which is commonly performed by repeated freeze and thaw cycles. Such culture supplements are notoriously ill-defined, and many parameters contribute to batch-to-batch variation in hPL such as different amounts of plasma, a broad range of growth factors and donor-specific effects. The plasma components of hPL necessitate addition of anticoagulants such as heparins to prevent gelatinization of hPL medium, and their concentration must be standardized. Labels for description of hPL-such as "xenogen-free," "animal-free" and "serum free"-are not used consistently in the literature and may be misleading if not critically assessed. Further analysis of the precise composition of relevant growth factors, attachment factors, microRNAs and exosomes will pave the way for optimized and defined culture conditions. The use of hPL has several advantages and disadvantages: they must be taken into account because the choice of cell culture additive has major impact on cell preparations. Copyright © 2014 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  4. Fetal bovine serum and human constitutive androstane receptor: Evidence for activation of the SV23 splice variant by artemisinin, artemether, and arteether in a serum-free cell culture system

    Energy Technology Data Exchange (ETDEWEB)

    Lau, Aik Jiang; Chang, Thomas K.H., E-mail: thomas.chang@ubc.ca

    2014-06-01

    The naturally occurring SV23 splice variant of human constitutive androstane receptor (hCAR-SV23) is activated by di-(2-ethylhexyl)phthalate (DEHP), which is detected as a contaminant in fetal bovine serum (FBS). In our initial experiment, we compared the effect of dialyzed FBS, charcoal-stripped, dextran-treated FBS (CS-FBS), and regular FBS on the basal activity and ligand-activation of hCAR-SV23 in a cell-based reporter gene assay. In transfected HepG2 cells cultured in medium supplemented with 10% FBS, basal hCAR-SV23 activity varied with the type of FBS (regular > dialyzed > CS). DEHP increased hCAR-SV23 activity when 10% CS-FBS, but not regular FBS or dialyzed FBS, was used. With increasing concentrations (1–10%) of regular FBS or CS-FBS, hCAR-SV23 basal activity increased, whereas in DEHP-treated cells, hCAR-SV23 activity remained similar (regular FBS) or slightly increased (CS-FBS). Subsequent experiments identified a serum-free culture condition to detect DEHP activation of hCAR-SV23. Under this condition, artemisinin, artemether, and arteether increased hCAR-SV23 activity, whereas they decreased it in cells cultured in medium supplemented with 10% regular FBS. By comparison, FBS increased the basal activity of the wild-type isoform of hCAR (hCAR-WT), whereas it did not affect the basal activity of the SV24 splice variant (hCAR-SV24) or ligand activation of hCAR-SV24 and hCAR-WT by 6-(4-chlorophenyl)imidazo[2,1-b][1,3]thiazole-5-carbaldehyde O-(3,4-dichlorobenzyl)oxime (CITCO). The use of serum-free culture condition was suitable for detecting CITCO activation of hCAR-WT and hCAR-SV24. In conclusion, FBS leads to erroneous classification of pharmacological ligands of hCAR-SV23 in cell-based assays, but investigations on functional ligands of hCAR isoforms can be conducted in serum-free culture condition. - Highlights: • FBS leads to erroneous pharmacological classification of hCAR-SV23 ligands. • Artemisinin, artemether, and arteether activate h

  5. Free serum thyroxine

    International Nuclear Information System (INIS)

    Degrossi, O.J.; Altschuler, Noe; Cabrejas, M.L. de; Pinkas, Mirta; Garcia del Rio, Hernan

    1982-01-01

    The use of radiommunoassay (RIA) tehcniques has increased the diagnosis of thyroid functional alterations. A solid phase RIA method for free thyroxine (FT4) measurement was tested. Serum FT4, Total T4, T3 and TSH were determined by radioimmunoassay in 179 subjects. One hundred twenty two patients were normal (8 to 75 years old); FT4 was 1.42 +- 0.03 ng/100 ml (avg. value and std. error). In 27 cases of thyrotoxicosis the values were 4.66 +- 0.48 ng/100 ml and in 15 cases of hypothyroidism 0.50 +- 0.06 ng/100 ml (statistics probability [es

  6. The down-regulation of the mitogenic fibrinogen receptor (MFR) in serum-containing medium does not occur in defined medium.

    Science.gov (United States)

    Levesque, J P; Hatzfeld, A; Domart, I; Hatzfeld, J

    1990-02-01

    Normal human hemopoietic cells such as early bone marrow progenitors, or lymphoma-derived cell lines such as Raji or JM cells, possess a low-affinity receptor specific for fibrinogen. This receptor triggers a mitogenic effect. It differs from the glycoprotein IIb-IIIa which is involved in fibrinogen-induced platelet aggregation. We demonstrate here that this mitogenic fibrinogen receptor (MFR) can be internalized or reexpressed, depending on culture conditions. Internalization was temperature-dependent. At 37 degrees C in the presence of cycloheximide or actinomycin D, the half-life of cell surface MFRs was 2 h, independent of receptor occupancy. Binding of fibrinogen to the MFR resulted in a down-regulation which was fibrinogen dose-dependent. This occurred in serum-supplemented medium but not in defined medium supplemented with fatty acids. Reexpression of MFRs could be induced in 28 to 42 h by serum removal. The down-regulation of mitogenic receptors in plasma or serum could explain why normal cells do not proliferate in the peripheral blood.

  7. An improved ultrafiltration method for determining free testosterone in serum

    International Nuclear Information System (INIS)

    Vlahos, I.; MacMahon, W.; Sgoutas, D.; Bowers, W.; Thompson, J.; Trawick, W.

    1982-01-01

    In this method, we use the Amicon MPS-1 centrifugal ultrafiltration device and the YMB membrane in measuring free testosterone in serum. Two independent assays are combined: total testosterone and the ultrafiltrable fraction of added [ 3 H]testosterone. The unbound fraction is determined in 0.15-0.5 mL ultrafiltrates of 0.6 to 1 mL of variably diluted serum that has been equilibrated with [ 3 H]testosterone at 37 degrees C. The assay is rapid (less than 1 h), practicable (requires 0.6 mL of serum), and reproducible (CV 3.2% within assay, 3.9% between assays). Accuracy was evaluated as the fraction of free testosterone in the ultrafiltrate of dialyzed serum vs that in a prior dialysate; they were the same confirming the validity of the free testosterone measurement. Samples from ostensibly healthy men and women and from hirsute and pregnant women gave results that agreed with those obtained by equilibrium dialysis. Total testosterone concentrations for normal and hirsute women showed considerable overlap, but data on free testosterone concentrations in these populations were better resolved

  8. Effects of serum on cytotoxicity of nano- and micro-sized ZnO particles

    Energy Technology Data Exchange (ETDEWEB)

    Hsiao, I-Lun; Huang, Yuh-Jeen, E-mail: yjhuang@mx.nthu.edu.tw [National Tsing Hua University, Department of Biomedical Engineering and Environmental Sciences (China)

    2013-09-15

    Although an increasing number of in vitro studies are being published regarding the cytotoxicity of nanomaterials, the components of the media for toxicity assays have often varied according to the needs of the scientists. Our aim for this study was to evaluate the influence of serum-in this case, fetal bovine serum-in a cell culture medium on the toxicity of nano-sized (50-70 nm) and micro-sized (<1 {mu}m) ZnO on human lung epithelial cells (A549). The nano- and micro-sized ZnO both exhibited their highest toxicity when exposed to serum-free media, in contrast to exposure in media containing 5 or 10 % serum. This mainly comes not only from the fact that ZnO particles in the serum-free media have a higher dosage-per-cell ratio, which results from large aggregates of particles, rapid sedimentation, absence of protein protection, and lower cell growth rate, but also that extracellular Zn{sup 2+} release contributes to cytotoxicity. Although more extracellular Zn{sup 2+} release was observed in serum-containing media, it did not contribute to nano-ZnO cytotoxicity. Furthermore, non-dissolved particles underwent size-dependent particle agglomeration, resulting in size-dependent toxicity in both serum-containing and serum-free media. A low correlation between cytotoxicity and inflammation endpoints in the serum-free medium suggested that some signaling pathways were changed or induced. Since cell growth, transcription behavior for protein production, and physicochemical properties of ZnO particles all were altered in serum-free media, we recommend the use of a serum-containing medium when evaluating the cytotoxicity of NPs.

  9. The B-domain of factor VIII reduces cell membrane attachement to host cells in serum free conditions

    DEFF Research Database (Denmark)

    Kolind, Mille Petersen; Nørby, Peder Lisby; Flintegaard, Thomas Veje

    2010-01-01

    engineered extensively throughout the years to increase the low production yields that initially were obtained from mammalian cell cultures. The scope of this work was to investigate the interaction of rFVIII with the cell membrane surface of the producing cells in serum free medium. We wondered whether...... binding of rFVIII to the cell membrane could be a factor diminishing the production yield. We studied the contribution of the rFVIII B-domain to membrane attachment by transfecting several constructs containing increasing lengths of the B-domain into cells under serum free conditions. We found that 90......% of rFVIII is attached to the cell membrane of the producing cell when the rFVIII variant contains a short B-domain (21 aa). By increasing the length of the B-domain the membrane attached fraction can be reduced to 50% of the total expressed rFVIII. Further, our studies show that the N...

  10. Multiple free-radical scavenging (MULTIS) capacity in cattle serum.

    Science.gov (United States)

    Sueishi, Yoshimi; Kamogawa, Erisa; Kimura, Anna; Kitahara, Go; Satoh, Hiroyuki; Asanuma, Taketoshi; Oowada, Shigeru

    2017-01-01

    Multiple free-radical scavenging (MULTIS) activity in cattle and human sera was evaluated with electron spin resonance spectroscopy. Scavenging rates against six active species, namely hydroxyl radical, superoxide anion, alkoxyl radical, alkylperoxyl radical, methyl radical, and singlet oxygen were quantified. The difference in the electron spin resonance signal intensity in the presence and absence of the serum was converted into the scavenging rates. Comparative MULTIS measurements were made in sera from eight beef cattle, three fetal calves and fifteen healthy human volunteers. Further, we determined the MULTIS value of albumin, the most abundant component in serum. MULTIS values in cattle sera indicated higher scavenging activity against most free radical species tested than human sera. In particular, cattle serum scavenging activities against superoxide and methyl radical were higher than human serum by 2.6 and 3.7 fold, respectively. In cattle serum, albumin appears to play a dominant role in MULTIS activity, but in human serum that is not the case. Previous data indicated that the abundance of uric acid in bovine blood is nearly 80% less than humans; however, this difference does not explain the deviation in MULTIS profile.

  11. Serum-free Erythroid Differentiation for Efficient Genetic Modification and High-Level Adult Hemoglobin Production.

    Science.gov (United States)

    Uchida, Naoya; Demirci, Selami; Haro-Mora, Juan J; Fujita, Atsushi; Raines, Lydia N; Hsieh, Matthew M; Tisdale, John F

    2018-06-15

    In vitro erythroid differentiation from primary human cells is valuable to develop genetic strategies for hemoglobin disorders. However, current erythroid differentiation methods are encumbered by modest transduction rates and high baseline fetal hemoglobin production. In this study, we sought to improve both genetic modification and hemoglobin production among human erythroid cells in vitro . To model therapeutic strategies, we transduced human CD34 + cells and peripheral blood mononuclear cells (PBMCs) with lentiviral vectors and compared erythropoietin-based erythroid differentiation using fetal-bovine-serum-containing media and serum-free media. We observed more efficient transduction (85%-93%) in serum-free media than serum-containing media (20%-69%), whereas the addition of knockout serum replacement (KSR) was required for serum-free media to promote efficient erythroid differentiation (96%). High-level adult hemoglobin production detectable by electrophoresis was achieved using serum-free media similar to serum-containing media. Importantly, low fetal hemoglobin production was observed in the optimized serum-free media. Using KSR-containing, serum-free erythroid differentiation media, therapeutic adult hemoglobin production was detected at protein levels with β-globin lentiviral transduction in both CD34 + cells and PBMCs from sickle cell disease subjects. Our in vitro erythroid differentiation system provides a practical evaluation platform for adult hemoglobin production among human erythroid cells following genetic manipulation.

  12. Salivary Cortisol Can Replace Free Serum Cortisol Measurements in Patients With Septic Shock

    Science.gov (United States)

    Orlander, Philip R.

    2011-01-01

    Background: There is a renewed interest in adrenal function during severe sepsis. Most studies have used total serum cortisol levels; however, only free serum cortisol is biologically active. The aim of this study was to determine the validity of salivary cortisol levels as a surrogate for free serum cortisol levels during septic shock. Methods: Fifty-seven patients with septic shock were studied to determine the correlation between total serum cortisol and salivary cortisol to free serum cortisol levels. Thirty-eight patients were included in the salivary to free serum cortisol correlation. Salivary cortisol level was tested by enzyme immunoassay. Serum total cortisol, free cortisol, and cortisol-binding globulin (CBG) levels were determined by liquid chromatography-mass spectrometry, equilibrium analysis, and radioimmunoassay, respectively. Results: The mean ± SD age was 56.6 ± 18.5 years. Fifty-seven percent were women. APACHE (Acute Physiology and Chronic Health Evaluation) II score median was 26, Simplified Acute Physiology Score II median was 61, and Sequential Organ Failure Assessment median was 13. The correlation between salivary and free serum cortisol levels was 0.79 (95% CI, 0.63-0.89; P cortisol and total serum cortisol levels was 0.86 (95% CI, 0.78-0.92; P cortisol level was 2.27 ± 1.64 μg/dL. The mean ± SD salivary cortisol level was 2.60 ± 2.69 μg/dL. The mean ± SD total serum cortisol level was 21.56 ± 8.71 μg/dL. The mean ± SD CBG level was 23.54 ± 8.33 mg/dL. Conclusions: Salivary cortisol level can be used as a surrogate of free serum cortisol level in patients with septic shock with very good correlation. Salivary cortisol testing is noninvasive, easy to perform, and can be conducted daily. Trial registry: ClinicalTrials.gov; No.: NCT00523198; URL: www.clinicaltrials.gov PMID:21816912

  13. Momentum anisotropy at freeze out

    International Nuclear Information System (INIS)

    Feld, S.; Borghini, N.; Lang, C.

    2017-01-01

    The transition from a hydrodynamical modeling to a particle-based approach is a crucial element of the description of high-energy heavy-ion collisions. Assuming this “freeze out” happens instantaneously at each point of the expanding medium, we show that the local phase-space distribution of the emitted particles is asymmetric in momentum space. This suggests the use of anisotropic hydrodynamics for the last stages of the fluid evolution. We discuss how observables depend on the amount of momentum-space anisotropy at freeze out and how smaller or larger anisotropies allow for different values of the freeze-out temperature. (paper)

  14. EPINEPHRINE CONCENTRATION IN WILD BOAR (SUS SCROFA L. SERUM AFTER REPEATED ELISA TESTED FREEZE-THAWING CYCLES

    Directory of Open Access Journals (Sweden)

    Neška Vukšić

    2013-06-01

    Full Text Available Blood samples for determining of epinephrine concentration and biochemical parameters in the blood serum of wild boars were taken from 42 healthy wild boars, both sexes, during the hunting season. All animals in good condition, body weight 20 to 95 kg, were divided into two groups up to 50 kg (group A and up to 95 kg (Group B. Epinephrine concentration was determined by ELISA twice: one week after taking samples and a month after repeated freezing at -80°C. It was higher in relation to the reference value of domestic pigs and human (109.45 pg/ml in A and 119.54 pg/ml in B group. Repeated freezing and re-analysis after a month were resulted in lower concentrations of epinephrine (12% in young and 11.17% in adult animal, but without statistical significance (P>0.05. Biochemical analysis results’ show increased glucose and triglycerides concentrations compared to the reference values, while other indicators were observed within or slightly increased referring to the normal range. The correlation between glucose and epinephrine was not determined.

  15. Serum-Free Media and the Immunoregulatory Properties of Mesenchymal Stem Cells In Vivo and In Vitro

    Directory of Open Access Journals (Sweden)

    Mei Wu

    2014-02-01

    Full Text Available Background: Mesenchymal stem cells are capable of self-renewal and multi-lineage differentiation. They are used extensively to treat several diseases. Traditionally, mesenchymal stem cells are cultured in serum-containing media, typically supplemented with fetal bovine serum (FBS. However, the variability of FBS is likely to skew experimental results. Although serum-free media used to expand mesenchymal stem cells has facilitated remarkable achievements, immunomodulation of these cells in under serum-free conditions is poorly understood. We hypothesized that mesenchymal stem cells expanded in serum-free media will retain powerful immunoregulatory functions in vitro and in vivo. Design and Methods: Immunosuppressive activity and the immunomodulatory cytokines produced by mesenchymal stem cells in serum-free media were characterized in vitro. Immunomodulation by serum-free mesenchymal stem cell expansion in monocrotaline-induced pulmonary hypertension was explored in vivo. Results: Similar to cells in serum-containing media, mesenchymal stem cells expanded in serum-free media inhibited proliferation and apoptosis of CD4+T cells. They also exhibited strong immunosuppressive activities and secreted high levels of immunomodulatory cytokines such as PGE2, IDO1, COX2, IL-6, and IL-1β, but not HGF. On the other hand, growth of mesenchymal stem cells in serum-free media attenuated pulmonary vascular remodeling and inhibited mRNA expression of proinflammatory cytokines TNF-α, IFN-γ, IL-6, IL-1β, and IL-18. Conclusions: Mesenchymal stem cells in serum-free media maintained powerful immunomodulatory function in vitro and in vivo; serum-free media may replace serum-containing media for basic research and clinical applications.

  16. Ultrasensitive radioimmunoassay for direct determination of free triiodothyronine concentration in serum

    International Nuclear Information System (INIS)

    Weeke, J.; Oerskov, H.

    1975-01-01

    Free triiodothyronine (T 3 ) in serum has been measured directly in dialysates of serum, using a wick chromatographic radioimmunoassay. Adequate sensitivity was attained by the use of [ 125 I]T 3 with a very high specific activity (2,000 to 3,000μCi/μg). Sera were dialysed against a Krebs-Ringer bicarbonate buffer modified so as to be similar to plasma water. Dialysis took place under carefully controlled circumstances. The influence on the equilibrium of total to free T 3 of temperature, serum dilution and dialysis time was studied. By the present method, free T 3 in serum from groups of subjects including 20 men, 10 women taking oral contraceptives and 20 women with normal menstrual cycles were identical, averaging 5.2 pg/ml. A chromatographic radioimmunoassay of total T 3 using high specific activity [ 125 I]T 3 and a very small test sample is also described. (auth.)

  17. Anhydrobiosis and Freezing-Tolerance

    DEFF Research Database (Denmark)

    McGill, Lorraine; Shannon, Adam; Pisani, Davide

    2015-01-01

    Anhydrobiotic animals can survive the loss of both free and bound water from their cells. While in this state they are also resistant to freezing. This physiology adapts anhydrobiotes to harsh environments and it aids their dispersal. Panagrolaimus davidi, a bacterial feeding anhydrobiotic nematode...... Panagrolaimus strains from tropical, temperate, continental and polar habitats and we analysed their phylogenetic relationships. We found that several other Panagrolaimus isolates can also survive freezing when fully hydrated and that tissue extracts from these freezing-tolerant nematodes can inhibit the growth...

  18. Attachment and growth of human keratinocytes in a serum-free environment.

    Science.gov (United States)

    Gilchrest, B A; Calhoun, J K; Maciag, T

    1982-08-01

    Using a serum-free system, we have investigated the influence of human fibronectin (HFN) and selected growth factors (GF) on the attachment and growth of normal human keratinocytes in vitro. Single-cell suspensions of keratinocytes from near-confluent primary plates, plated on 5-10 microgram/cm2 HFN, showed approximately 30-40% attachment after 2-24 hours of incubation at 37 degrees C, compared with 4-6% attachment on uncoated platic plates. Percentage of attached cells was independent of seed density, tissue donor age, in vitro culture age, or medium composition, while subsequent cellular proliferation was strongly dependent on these factors. Keratinocytes grown on an adequate HFN matrix in a previously described hormone-supplemented medium (Maciag et al., 1981a) achieved four to eight population doubling over 7-12 days at densities greater than or equal to 104 cell/cm2. Removal of most GF individually from the medium had little or no effect on growth, while removal of epidermal growth factor (EGF) alone reduced growth by 30-35% and removal of bovine brain extract (BE) alone reduced growth by approximately 90%. Conversely, EGF alone in basal medium supported approximately 10% control growth, BE alone supported 30-40% control growth, and the combination of EGF and BE approximately 70%. In addition to its major effect on proliferation in this system, BE was necessary to preserve normal keratinocyte morphology and protein production. These findings expand earlier observations that HFN facilitates keratinocyte attachment in vitro and that a brain-derived extract can exert a major positive influence on cultured keratinocytes.

  19. Multiple free-radical scavenging capacity in serum

    Science.gov (United States)

    Oowada, Shigeru; Endo, Nobuyuki; Kameya, Hiromi; Shimmei, Masashi; Kotake, Yashige

    2012-01-01

    We have developed a method to determine serum scavenging-capacity profile against multiple free radical species, namely hydroxyl radical, superoxide radical, alkoxyl radical, alkylperoxyl radical, alkyl radical, and singlet oxygen. This method was applied to a cohort of chronic kidney disease patients. Each free radical species was produced with a common experimental procedure; i.e., uv/visible-light photolysis of free-radical precursor/sensitizer. The decrease in free-radical concentration by the presence of serum was quantified with electron spin resonance spin trapping method, from which the scavenging capacity was calculated. There was a significant capacity change in the disease group (n = 45) as compared with the healthy control group (n = 30). The percent values of disease’s scavenging capacity with respect to control group indicated statistically significant differences in all free-radical species except alkylperoxyl radical, i.e., hydroxyl radical, 73 ± 12% (p = 0.001); superoxide radical, 158 ± 50% (p = 0.001); alkoxyl radical, 121 ± 30% (p = 0.005); alkylperoxyl radical, 123 ± 32% (p>0.1); alkyl radical, 26 ± 14% (p = 0.001); and singlet oxygen, 57 ± 18% (p = 0.001). The scavenging capacity profile was illustrated using a radar chart, clearly demonstrating the characteristic change in the disease group. Although the cause of the scavenging capacity change by the disease state is not completely understood, the profile of multiple radical scavenging capacities may become a useful diagnostic tool. PMID:22962529

  20. Universal pion freeze-out in heavy-ion collisions.

    Science.gov (United States)

    Adamová, D; Agakichiev, G; Appelshäuser, H; Belaga, V; Braun-Munzinger, P; Castillo, A; Cherlin, A; Damjanović, S; Dietel, T; Dietrich, L; Drees, A; Esumi, S I; Filimonov, K; Fomenko, K; Fraenkel, Z; Garabatos, C; Glässel, P; Hering, G; Holeczek, J; Kushpil, V; Lenkeit, B; Ludolphs, W; Maas, A; Marín, A; Milosević, J; Milov, A; Miśkowiec, D; Panebrattsev, Yu; Petchenova, O; Petrácek, V; Pfeiffer, A; Rak, J; Ravinovich, I; Rehak, P; Sako, H; Schmitz, W; Schukraft, J; Sedykh, S; Shimansky, S; Slívová, J; Specht, H J; Stachel, J; Sumbera, M; Tilsner, H; Tserruya, I; Wessels, J P; Wienold, T; Windelband, B; Wurm, J P; Xie, W; Yurevich, S; Yurevich, V

    2003-01-17

    Based on an evaluation of data on pion interferometry and on particle yields at midrapidity, we propose a universal condition for thermal freeze-out of pions in heavy-ion collisions. We show that freeze-out occurs when the mean free path of pions lambda(f) reaches a value of about 1 fm, which is much smaller than the spatial extent of the system at freeze-out. This critical mean free path is independent of the centrality of the collision and beam energy from the Alternating Gradient Synchrotron to the Relativistic Heavy Ion Collider.

  1. Establishing elements of a synthetic biology platform for Vaccinia virus production: BioBrick™ design, serum-free virus production and microcarrier-based cultivation of CV-1 cells.

    Science.gov (United States)

    Liu, Shuchang; Ruban, Ludmila; Wang, Yaohe; Zhou, Yuhong; Nesbeth, Darren N

    2017-02-01

    Vaccinia virus (VACV) is an established vector for vaccination and is beginning to prove effective as an oncolytic agent. Industrial production of VACV stands to benefit in future from advances made by synthetic biology in genome engineering and standardisation. The CV-1 cell line can be used for VACV propagation and has been used extensively with the CRISPR/Cas9 system for making precise edits of the VACV genome. Here we take first steps toward establishing a scalable synthetic biology platform for VACV production with CV-1 cells featuring standardised biological tools and serum free cell cultivation. We propose a new BioBrick™ plasmid backbone format for inserting transgenes into VACV. We then test the performance of CV-1 cells in propagation of a conventional recombinant Lister strain VACV, VACVL-15 RFP, in a serum-free process. CV-1 cells grown in 5% foetal bovine serum (FBS) Dulbecco's Modified Eagle Medium (DMEM) were adapted to growth in OptiPRO and VP-SFM brands of serum-free media. Specific growth rates of 0.047 h -1 and 0.044 h -1 were observed for cells adapted to OptiPRO and VP-SFM respectively, compared to 0.035 h -1 in 5% FBS DMEM. Cells adapted to OptiPRO and to 5% FBS DMEM achieved recovery ratios of over 96%, an indication of their robustness to cryopreservation. Cells adapted to VP-SFM showed a recovery ratio of 82%. Virus productivity in static culture, measured as plaque forming units (PFU) per propagator cell, was 75 PFU/cell for cells in 5% FBS DMEM. VP-SFM and OptiPRO adaptation increased VACV production to 150 PFU/cell and 350 PFU/cell respectively. Boosted PFU/cell from OptiPRO-adapted cells persisted when 5% FBS DMEM or OptiPRO medium was observed during the infection step and when titre was measured using cells adapted to 5% FBS DMEM or OptiPRO medium. Finally, OptiPRO-adapted CV-1 cells were successfully cultivated using Cytodex-1 microcarriers to inform future scale up studies.

  2. Establishing elements of a synthetic biology platform for Vaccinia virus production: BioBrick™ design, serum-free virus production and microcarrier-based cultivation of CV-1 cells

    Directory of Open Access Journals (Sweden)

    Shuchang Liu

    2017-02-01

    Full Text Available Vaccinia virus (VACV is an established vector for vaccination and is beginning to prove effective as an oncolytic agent. Industrial production of VACV stands to benefit in future from advances made by synthetic biology in genome engineering and standardisation. The CV-1 cell line can be used for VACV propagation and has been used extensively with the CRISPR/Cas9 system for making precise edits of the VACV genome. Here we take first steps toward establishing a scalable synthetic biology platform for VACV production with CV-1 cells featuring standardised biological tools and serum free cell cultivation. We propose a new BioBrick™ plasmid backbone format for inserting transgenes into VACV. We then test the performance of CV-1 cells in propagation of a conventional recombinant Lister strain VACV, VACVL-15 RFP, in a serum-free process. CV-1 cells grown in 5% foetal bovine serum (FBS Dulbecco’s Modified Eagle Medium (DMEM were adapted to growth in OptiPRO and VP-SFM brands of serum-free media. Specific growth rates of 0.047 h−1 and 0.044 h−1 were observed for cells adapted to OptiPRO and VP-SFM respectively, compared to 0.035 h−1 in 5% FBS DMEM. Cells adapted to OptiPRO and to 5% FBS DMEM achieved recovery ratios of over 96%, an indication of their robustness to cryopreservation. Cells adapted to VP-SFM showed a recovery ratio of 82%. Virus productivity in static culture, measured as plaque forming units (PFU per propagator cell, was 75 PFU/cell for cells in 5% FBS DMEM. VP-SFM and OptiPRO adaptation increased VACV production to 150 PFU/cell and 350 PFU/cell respectively. Boosted PFU/cell from OptiPRO-adapted cells persisted when 5% FBS DMEM or OptiPRO medium was observed during the infection step and when titre was measured using cells adapted to 5% FBS DMEM or OptiPRO medium. Finally, OptiPRO-adapted CV-1 cells were successfully cultivated using Cytodex-1 microcarriers to inform future scale up studies.

  3. Verification of serum reference intervals for free light chains in a local South African population.

    Science.gov (United States)

    Zemlin, Annalise E; Rensburg, Megan A; Ipp, Hayley; Germishuys, Jurie J; Erasmus, Rajiv T

    2013-11-01

    Monoclonal serum free light chain measurements are used to follow up and manage patients with monoclonal gammopathies, and abnormal serum free light chain ratios are associated with risk of progression in certain diseases. We aimed to validate the reference intervals in our population. Reference intervals for κ and λ free light chains were established on 120 healthy adults. Creatinine levels were measured to exclude renal dysfunction and serum protein electrophoresis was performed. All creatinine values were within normal limits. After exclusion of subjects with abnormal serum protein electrophoreses, 113 subjects were available for analysis. The 95% reference interval was 6.3-20.6 mg/L for κ free light chains, 8.7-25.9 mg/L for λ free light chains and 0.46-1.23 for free light chain ratio. Most of the values fell within the manufacturer's recommended limits and therefore could be used for our population.

  4. Effects of Macromolecular Crowding on Human Adipose Stem Cell Culture in Fetal Bovine Serum, Human Serum, and Defined Xeno-Free/Serum-Free Conditions.

    Science.gov (United States)

    Patrikoski, Mimmi; Lee, Michelle Hui Ching; Mäkinen, Laura; Ang, Xiu Min; Mannerström, Bettina; Raghunath, Michael; Miettinen, Susanna

    2017-01-01

    Microenvironment plays an important role for stem cell proliferation and differentiation. Macromolecular crowding (MMC) was recently shown to assist stem cells in forming their own matrix microenvironment in vitro. The ability of MMC to support adipose stem cell (ASC) proliferation, metabolism, and multilineage differentiation was studied under different conditions: fetal bovine serum- (FBS-) and human serum- (HS-) based media and xeno- and serum-free (XF/SF) media. Furthermore, the immunophenotype of ASCs under MMC was evaluated. The proliferative capacity of ASCs under MMC was attenuated in each condition. However, osteogenic differentiation was enhanced under MMC, shown by increased deposition of mineralized matrix in FBS and HS cultures. Likewise, significantly greater lipid droplet accumulation and increased collagen IV deposition indicated enhanced adipogenesis under MMC in FBS and HS cultures. In contrast, chondrogenic differentiation was attenuated in ASCs expanded under MMC. The ASC immunophenotype was maintained under MMC with significantly higher expression of CD54. However, MMC impaired metabolic activity and differentiation capacity of ASCs in XF/SF conditions. Both the supportive and inhibitory effects of MMC on ASC are culture condition dependent. In the presence of serum, MMC maintains ASC immunophenotype and enhances adipogenic and osteogenic differentiation at the cost of reduced proliferation.

  5. Free testosterone as marker of adaptation to medium-intensive exercise.

    Science.gov (United States)

    Shkurnikov, M U; Donnikov, A E; Akimov, E B; Sakharov, D A; Tonevitsky, A G

    2008-09-01

    A 4-week study of adaptation reserves of the body was carried out during medium intensive exercise (medium intensive training: 60-80% threshold anaerobic metabolism). Two groups of athletes were singled out by the results of pulsometry analysis: with less than 20% work duration at the level above the 80% threshold anaerobic metabolism and with more than 20% work duration at the level above 80% threshold anaerobic metabolism. No appreciable differences between the concentrations of total testosterone, growth hormone, and cortisol before and after exercise in the groups with different percentage of anaerobic work duration were detected. In group 1 the concentrations of free testosterone did not change throughout the period of observation in comparison with the levels before training. In group 2, the level of free testosterone increased in comparison with the basal level: from 0.61+/-0.12 nmol/liter at the end of week 1 to 0.98+/-0.11 nmol/liter at the end of week 4 (p<0.01). The results indicate that the level of free testosterone can be used for evaluating the degree of athlete's adaptation to medium intensive exercise.

  6. Universal pion freeze-out in heavy-ion collisions

    CERN Document Server

    Adamova, D; Appelshäuser, H; Belaga, V; Braun-Munzinger, P; Castillo, A; Cherlin, A; Damjanovic, S; Dietel, T; Dietrich, L; Drees, A; Esumi, S I; Filimonov, K; Fomenko, K; Fraenkel, Zeev; Garabatos, C; Glässel, P; Hering, G; Holeczek, J; Kushpil, V; Lenkeit, B C; Ludolphs, W; Maas, A; Marin, A; Milosevic, J; Milov, A; Miskowiec, D; Panebratsev, Yu A; Petchenova, O Yu; Petracek, V; Pfeiffer, A; Rak, J; Ravinovich, I; Rehak, P; Sako, H; Schükraft, Jürgen; Sedykh, S; Shimansky, S S; Slivova, J; Specht, H J; Stachel, J; Sumbera, M; Tilsner, H; Tserruya, Itzhak; Wessels, J P; Wienold, T; Windelband, B; Wurm, J P; Xie, W; Yurevich, S; Yurevich, V; Schmitz, W

    2003-01-01

    Based on an evaluation of recent systematic data on two-pion interferometry and on measured particle yields at mid-rapidity, we propose a universal condition for thermal freeze-out of pions in heavy-ion collisions. We show that freeze-out occurs when the mean free path of pions lambda_f reaches a value of approximately 2.5 fm, which is much smaller than the spatial extent of the system at freeze-out. This critical mean free path is independent of the centrality of the collision and its value is constant at all currently available beam energies from AGS to RHIC.

  7. The replacement of serum by hormones in cell culture media.

    Science.gov (United States)

    Sato, G; Hayashi, I

    1976-12-01

    The replacement of serum by hormones in cell culture media. (Reemplazo del suero por hormonas en el medio de cultivo de células). Arch. Biol. Med. Exper. 10: 120-121, 1976. The serum used in cell culture media can be replaced by a mixture of hormones and some accesory blood factors. The pituitary cell line GH3 can be grown in a medium in which serum is replaced by triiodothyronine, transferrin, parathormone, tyrotrophin releasing hormone and somatomedins. Hela and BHK cell strains can also be grown in serum free medium supplemented with hormones. Each cell type appears to have different hormonal requirements yet it may found that some hormones are required for most cell types.

  8. A low percentage of autologous serum can replace bovine serum to engineer human nasal cartilage

    Directory of Open Access Journals (Sweden)

    F Wolf

    2008-02-01

    Full Text Available For the generation of cell-based therapeutic products, it would be preferable to avoid the use of animal-derived components. Our study thus aimed at investigating the possibility to replace foetal bovine serum (FBS with autologous serum (AS for the engineering of cartilage grafts using expanded human nasal chondrocytes (HNC. HNC isolated from 7 donors were expanded in medium containing 10% FBS or AS at different concentrations (2%, 5% and 10% and cultured in pellets using serum-free medium or in Hyaff®-11 meshes using medium containing FBS or AS. Tissue forming capacity was assessed histologically (Safranin O, immunohistochemically (type II collagen and biochemically (glycosaminoglycans -GAG- and DNA. Differences among experimental groups were assessed by Mann Whitney tests. HNC expanded under the different serum conditions proliferated at comparable rates and generated cartilaginous pellets with similar histological appearance and amounts of GAG. Tissues generated by HNC from different donors cultured in Hyaff®-11 had variable quality, but the accumulated GAG amounts were comparable among the different serum conditions. Staining intensity for collagen type II was consistent with GAG deposition. Among the different serum conditions tested, the use of 2% AS resulted in the lowest variability in the GAG contents of generated tissues. In conclusion, a low percentage of AS can replace FBS both during the expansion and differentiation of HNC and reduce the variability in the quality of the resulting engineered cartilage tissues.

  9. Serum Immunoglobulin Free Light Chain Assessment in IgG4-Related Disease

    Directory of Open Access Journals (Sweden)

    Aurélie Grados

    2013-01-01

    Full Text Available Immunoglobulin free light chains are produced in excess during normal antibody synthesis. Their evaluation is commonly used in case of a monoclonal gammopathy. In polyclonal hypergammaglobulinemia related to the Sjögren syndrome or systemic lupus, erythematosus serum free light chain levels are increased and could correlate with disease activity. We show here that the κ ( and λ ( free light chains and the κ : λ ratio ( are increased in sixteen patients with IgG4-related disease when compared to healthy controls. The increase of κ and λ free light chains probably reflects the marked polyclonal B cell activation of the disease. We could not assess in this small cohort of patients a significative correlation of serum free light chain levels and disease activity or extension.

  10. A simplified ultrafiltration method for determination of serum free cortisol

    International Nuclear Information System (INIS)

    MacMahon, W.; Sgoutas, D.

    1983-01-01

    The authors describe the suitability of the Amicon MPS-1 centrifugal ultrafiltration device and the YMB membrane for measuring free cortisol in serum. The method combines two independent assays: total cortisol and the ultrafiltrate fraction of added [ 3 H]cortisol. The unbound fraction is determined in 0.25-0.30 ml of ultrafiltrate collected from 0.6 to 1 ml of serum that has been equilibrated with [ 3 H]cortisol at 37 0 C for 20 min. The assay is rapid (less than 1 h), practical (no more than 0.6 ml of serum is necessary) and repeatable (CV: 3.8% within-assay and 12.2% in different assays). Error introduced in free cortisol measurement due to dilution effects in dialysis is systematically defined, and the effect of tracer purity on the ultrafiltration method is examined. Dialyzed sera from normal men and women, from patients with Cushing's disease and adrenal insufficiency, and from pregnant women gave ultrafiltration results that accurately duplicated those obtained by previous dialysis. (Auth.)

  11. Human platelet lysate: Replacing fetal bovine serum as a gold standard for human cell propagation?

    Science.gov (United States)

    Burnouf, Thierry; Strunk, Dirk; Koh, Mickey B C; Schallmoser, Katharina

    2016-01-01

    The essential physiological role of platelets in wound healing and tissue repair builds the rationale for the use of human platelet derivatives in regenerative medicine. Abundant growth factors and cytokines stored in platelet granules can be naturally released by thrombin activation and clotting or artificially by freeze/thaw-mediated platelet lysis, sonication or chemical treatment. Human platelet lysate prepared by the various release strategies has been established as a suitable alternative to fetal bovine serum as culture medium supplement, enabling efficient propagation of human cells under animal serum-free conditions for a multiplicity of applications in advanced somatic cell therapy and tissue engineering. The rapidly increasing number of studies using platelet derived products for inducing human cell proliferation and differentiation has also uncovered a considerable variability of human platelet lysate preparations which limits comparability of results. The main variations discussed herein encompass aspects of donor selection, preparation of the starting material, the possibility for pooling in plasma or additive solution, the implementation of pathogen inactivation and consideration of ABO blood groups, all of which can influence applicability. This review outlines the current knowledge about human platelet lysate as a powerful additive for human cell propagation and highlights its role as a prevailing supplement for human cell culture capable to replace animal serum in a growing spectrum of applications. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Impact of nucleon mass shift on the freeze-out process

    International Nuclear Information System (INIS)

    Zschocke, Sven; Csernai, Laszlo Pal; Molnar, Etele; Nyiri, Agnes; Manninen, Jaakko

    2005-01-01

    The freeze-out of a massive nucleon gas through a finite layer with a timelike normal is studied. The impact of the in-medium nucleon mass shift on the freeze-out process is investigated. A considerable modification of the thermodynamic variables of temperature, flow velocity, energy density, and particle density has been found. Because of the nucleon mass shift the freeze-out particle distribution functions are changed noticeably in comparison with the evaluations, which use the vacuum nucleon mass

  13. Optimization of Serum Immunoglobulin Free Light Chain Analysis for Subclassification of Cardiac Amyloidosis.

    Science.gov (United States)

    Halushka, Marc K; Eng, George; Collins, A Bernard; Judge, Daniel P; Semigran, Marc J; Stone, James R

    2015-06-01

    Accurate and rapid classification of cardiac amyloidosis is important for patient management. We have optimized the use of serum free light chain kappa and lambda values to differentiate immunoglobulin light chain amyloid (AL) amyloidosis from transthyretin amyloid and amyloid A using 85 cases of tissue-proven cardiac amyloidosis, in which there was direct classification of amyloidosis by mass spectrometry or immunofluorescence. The serum free light chain kappa/lambda ratios were non-overlapping for the three major groups: AL-lambda (0.01-0.41, n = 30), non-AL (0.52-2.7, n = 43), and AL-kappa (6.7-967, n = 12). A kappa/lambda ratio value between 0.5 and 5.0 had 100 % sensitivity and 100 % specificity for distinguishing AL amyloidosis from non-AL amyloidosis. This optimized range for serum light chain kappa/lambda ratio provides extremely robust classification of cardiac amyloidosis. Cases of cardiac amyloidosis in which the serum kappa/lambda free light chain ratio falls close to these new cutoff values may benefit most from direct amyloid subtyping.

  14. Clinical significance of the measurements of serum free thyroxine and free triiodothyronine concentrations

    International Nuclear Information System (INIS)

    Kubota, Ken; Sasaki, Norio; Takaku, Fumimaro; Uchimura, Hidemasa

    1988-01-01

    A commercially available ''DPC'' radioimmunoassay kit was used to study the serum concentrations of free triiodothyronine (FT3) and free thyroxine (FT4) in a series of 189 patients with various thyroid diseases and 120 healthy controls. The basal serum concentrations of FT3 and FT4 in normal controls ranged from 0.98 to 2.04 ng/dl and from 1.43 to 3.66 pg/ml, respectively. All untreated patients with Graves' disease had abnormally high FT3 and FT4 values, indicating the discrimination between hyperthyroid and normal subjects. A decreased ratio of FT4 to FT3 was observed in patients managed with antithyroid drugs. In diagnosing hypothyroidism, ''DPC'' FT4 kits were more sensitive than ''DPC'' TT4 and ''Amerlex'' FT4 kits. In the case of non-thyroid diseases, FT4 and FT3 values, as well as total T4 and total T3, were normal or decreased, with the exception of occasionally unknown high values. The interference of thyroxine binding globulin abnormablity was successfully eliminated by using new versions of ''DPC'' kits. (Namekawa, K.)

  15. A protocol describing the use of a recombinant protein-based, animal product-free medium (APEL) for human embryonic stem cell differentiation as spin embryoid bodies.

    Science.gov (United States)

    Ng, Elizabeth S; Davis, Richard; Stanley, Edouard G; Elefanty, Andrew G

    2008-01-01

    In order to promote the uniform and reproducible differentiation of human embryonic stem cells (HESCs) in response to exogenously added growth factors, we have developed a method (spin embryoid bodies (EBs)) that uses a recombinant protein-based, animal product-free medium in which HESCs are aggregated by centrifugation to form EBs. In this protocol we describe the formulation of this medium, denoted APEL (Albumin Polyvinylalcohol Essential Lipids), and its use in spin EB differentiation of HESCs. We also describe a more economical variant, BPEL (Bovine Serum Albumin (BSA) Polyvinylalchohol Essential Lipids), in which BSA replaces the recombinant human albumin. The integration of a medium that includes only defined and recombinant components with a defined number of cells to initiate EB formation results in a generally applicable, robust platform for growth factor-directed HESC differentiation.

  16. Two different serum-free media and osmolality effect upon human 293 cell growth and adenovirus production.

    Science.gov (United States)

    Ferreira, Tiago B; Ferreira, Ana L; Carrondo, Manuel J T; Alves, Paula M

    2005-11-01

    Adenoviruses are promising vectors for gene therapy and vaccination protocols. Consequently, the market demands for adenovirus are increasing, driving the search for new methodologies for large-scale production of concentrated vectors with warranted purity and efficacy, in a cost-effective way. Nevertheless, the production of adenovirus is currently limited by the so-called 'cell density effect', i.e. a drop in cell specific productivity concomitant with increased cell concentration at infection. Of two different serum-free culture media (CD293 and EX-Cell), evaluated for their effect on human 293 cells growth and adenovirus production at cell densities higher than 1x10(6) cells/ml, EX-Cell proved the better medium for cell growth. Although adenovirus production was equivalent in both media when the infection was performed at 1x10(6) cells/ml, at 3x10(6) cells/ml CD293 was the better. This result related to the high ammonia content in EX-Cell medium at the highest cell concentration at infection. Besides this, the large-scale production of these vectors at high cell densities often requires re-feed strategies, which increase medium osmolality. While a negative effect on cell growth was observed with increasing osmolalities, adenovirus productivity was only affected for osmolalities higher than 430 mOsm.

  17. Elevated serum free thyroxine by thyroxine analog radioimmunoassays in euthyroid patients with familial dysalbuminemic hyperthyroxinemia

    International Nuclear Information System (INIS)

    Rajatanavin, R.; Fournier, L.; DeCosimo, D.; Abreau, C.; Braverman, L.E.

    1982-01-01

    A study was done to ascertain whether the serum free T4 measured by free T4 radioimmunoassay kits would, like equilibrium dialysis, be normal in patients with familial dysalbuminemic hyperthyroxinemia. Five free T4 radioimmunoassay kits were used to measure free T4 in serum samples from 19 patients with familial dysalbuminemic hyperthyroxinemia and 20 healthy volunteers. Values (mean +/- SE) for T4, free T4 index, and free T4 (equilibrium dialysis) in these normal subjects and patients with familial dysalbuminemic hyperthyroxinemia, respectively, were as follows: T4, 8.1 +/- 0.2 and 18.3 +/- 0.7 μg/dL; free T4 index, 3.1 +/- 0.1 and 7.3 +/- 0.3 μg/dL; free T4, 1.4 +/- 0.1 and 1.2 +/- 0.1 ng/dL. The following free T4 radioimmunoassay methods were used: antibody coated microfine silica, microencapsulated antibody, two-step antibody-coated tube, and one-step 125 I-T4 analog (2 kits). The present findings in patients with familial dysalbuminemic hyperthyroxinemia and previous observations in ill euthyroid patients suggest that serum free T4 measured by some radioimmunoassay methods must be interpreted with caution in these two clinical situations

  18. Storage of human platelets by freezing

    Energy Technology Data Exchange (ETDEWEB)

    Kim, B K; Tanoue, K; Baldini, M G

    1976-01-01

    Prolonged, probably indefinite storage of viable and functional human platelets is now possible by freezing with dimethylsulfoxide (DMSO). The platelets have a nearly normal survival upon reinfusion and are capable of sustained hemostatic effectiveness in thrombocytopenic patients. Adaptation of the freezing technique for large-scale usage has more recently been achieved. The method is mainly based on the following principles: (1) use of plasma for suspension of the platelet concentrate; (2) gradual addition (0.5% every 2 min) of DMSO to a final concentration of 5% and its gradual removal; (3) a slow cooling rate of about 1/sup 0/C per min and rapid thawing (in 1 min); (4) use of a polyolefin plastic bag for freezing; (5) a washing medium of 20% plasma in Hanks' balanced salt solution; (6) final resuspension of the platelets in 50% plasma in Hanks' solution.

  19. Higher Serum Levels of Free ĸ plus λ Immunoglobulin Light Chains Ameliorate Survival of Hemodialysis Patients

    DEFF Research Database (Denmark)

    Thilo, Florian; Caspari, Christina; Scholze, Alexandra

    2011-01-01

    Background/Aims: Impaired immune function is common in patients with chronic renal failure. Now, we determined whether serum levels of free immunoglobulin light chains predict mortality in patients with chronic kidney disease stage 5 on hemodialysis. Methods: We performed a prospective cohort study...... of 160 hemodialysis patients with a median follow-up of 15 months (interquartile range, 3-44 months). Serum levels of free κ and λ immunoglobulin light chains were measured at the start of the study. The primary end point was mortality from any cause. Results: In survivors, median serum levels of free κ...... plus λ immunoglobulin light chains were significantly higher compared with nonsurvivors (p light chains above the median compared with patients with serum levels below the median of 210 mg...

  20. Comparison of serum fractionation methods by data independent label-free proteomics

    Directory of Open Access Journals (Sweden)

    D. Baiwir

    2015-12-01

    Full Text Available Off-line sample prefractionations applied prior to biomarker discovery proteomics are options to enable more protein identifications and detect low-abundance proteins. This work compared five commercial methods efficiency to raw serum analysis using label-free proteomics. The variability of the protein quantities determined for each process was similar to the unprefractionated serum. A 49% increase in protein identifications and 12.2% of reliable quantification were obtained. A 61 times lower limit of protein quantitation was reached compared to protein concentrations observed in raw serum. The concentrations of detected proteins were confronted to estimated reference values.

  1. Can serum free fatty acids assessment predict severe preeclampsia?

    African Journals Online (AJOL)

    Nermeen Saad El Beltagy

    2011-10-20

    Oct 20, 2011 ... Methods: Twenty cases with severe preeclampsia (blood pressure P 160/110 after 20th week of ges- tation and ... ing factor with preeclampsia in non-obese pregnant women. ... Preeclampsia (PE) is a common pregnancy disorder that is ... centration of free fatty acids in the serum was measured by an.

  2. Comparison of serum hormone levels of captive and free-living maned wolves Chrysocyon brachyurus

    Directory of Open Access Journals (Sweden)

    O.B. Maia

    2008-02-01

    Full Text Available Serum hormone levels were compared between captive and free-living maned wolves and seasonal variations of sex hormones were studied. Blood samples were collected from 16 male and 26 female adult animals from Brazilian zoos, and from 30 male and 24 female free-living adults to determine serum progesterone and testosterone by radioimmunoassay. Serum testosterone concentrations varied (P 0.05. Sixteen captive males showed higher testosterone concentration during winter and spring compared with 30 free-living animals (P < 0.05. Progesterone concentration varied among seasons in 26 captive females (P < 0.05, being higher in autumn (15.3 ± 3.1 ng/mL than in summer (6.6 ± 1.5 ng/mL, winter (5.3 ± 3.1 ng/mL and spring (4.3 ± 0.7 ng/mL. Progesterone concentration of 24 free-living females varied between autumn (17.1 ± 6.0 ng/mL and winter (1.7 ± 0.3 ng/mL (P < 0.05, but we could not obtain data for spring or summer. No difference in progesterone levels was observed between captive and free-living females in autumn and winter.

  3. Opium consumption is negatively associated with serum prostate-specific antigen (PSA), free PSA, and percentage of free PSA levels.

    Science.gov (United States)

    Safarinejad, Mohammad Reza; Asgari, Seyyed Alaeddin; Farshi, Alireza; Iravani, Shahrokh; Khoshdel, Alireza; Shekarchi, Babak

    2013-01-01

    Addiction to opium continues to be a major worldwide medical and social problem. The study addressing the association between opium consumption and serum prostate-specific antigen (PSA) level is lacking. We determined the effects of opium consumption on serum PSA levels in opium-addict men. Our study subjects comprised 438 opium-addict men with a mean age of 52.2 ± 6.4 years (group 1). We compared these men with 446 men who did not indicate current or past opium use (group 2). Serum total PSA (tPSA), free PSA (fPSA), % fPSA, and sex hormones were compared between the 2 groups. The mean serum tPSA level was significantly lower in group 1 (1.05 ng/mL) than in controls (1.45 ng/mL) (P = 0.001). Opium consumption was also associated with lower fPSA (P = 0.001) and % fPSA (P = 0.001). Serum free testosterone level in opium-addict patients (132.5 ± 42 pg/mL) was significantly lower than that in controls (156.2 ± 43 pg/mL) (P = 0.03). However, no significant correlation existed between tPSA and free testosterone levels (r = 0.28, 95% CI, -0.036 to 0.51, P = 0.34). Among the patients with cancer in group 1, 35% were found to have high-grade tumor (Gleason score ≥ 7) compared with 26.7% in group 2 (P = 0.02). Total PSA and fPSA were strongly correlated with duration of opium use (r = -0.06, 95% CI, -0.04 to -0.08, P = 0.0001; and r = -0.05, 95% CI, -0.03 to -0.07, P = 0.0001, respectively). Opium consumption is independently and negatively associated with serum tPSA, fPSA, and % fPSA levels.

  4. Changes in Serum Free Amino Acids and Muscle Fatigue Experienced during a Half-Ironman Triathlon.

    Directory of Open Access Journals (Sweden)

    Francisco Areces

    Full Text Available The aim of this study was to investigate the relationship between changes in serum free amino acids, muscle fatigue and exercise-induced muscle damage during a half-ironman triathlon. Twenty-six experienced triathletes (age = 37.0 ± 6.8 yr; experience = 7.4 ± 3.0 yr competed in a real half-ironman triathlon in which sector times and total race time were measured by means of chip timing. Before and after the race, a countermovement jump and a maximal isometric force test were performed, and blood samples were withdrawn to measure serum free amino acids concentrations, and serum creatine kinase levels as a blood marker of muscle damage. Total race time was 320 ± 37 min and jump height (-16.3 ± 15.2%, P 20%. However, neither the changes in serum free amino acids nor the tryptophan/BCAA ratio were related muscle fatigue or muscle damage during the race.

  5. Expression of virulence factors by Staphylococcus aureus grown in serum.

    Science.gov (United States)

    Oogai, Yuichi; Matsuo, Miki; Hashimoto, Masahito; Kato, Fuminori; Sugai, Motoyuki; Komatsuzawa, Hitoshi

    2011-11-01

    Staphylococcus aureus produces many virulence factors, including toxins, immune-modulatory factors, and exoenzymes. Previous studies involving the analysis of virulence expression were mainly performed by in vitro experiments using bacterial medium. However, when S. aureus infects a host, the bacterial growth conditions are quite different from those in a medium, which may be related to the different expression of virulence factors in the host. In this study, we investigated the expression of virulence factors in S. aureus grown in calf serum. The expression of many virulence factors, including hemolysins, enterotoxins, proteases, and iron acquisition factors, was significantly increased compared with that in bacterial medium. In addition, the expression of RNA III, a global regulon for virulence expression, was significantly increased. This effect was partially restored by the addition of 300 μM FeCl₃ into serum, suggesting that iron depletion is associated with the increased expression of virulence factors in serum. In chemically defined medium without iron, a similar effect was observed. In a mutant with agr inactivated grown in serum, the expression of RNA III, psm, and sec4 was not increased, while other factors were still induced in the mutant, suggesting that another regulatory factor(s) is involved. In addition, we found that serum albumin is a major factor for the capture of free iron to prevent the supply of iron to bacteria grown in serum. These results indicate that S. aureus expresses virulence factors in adaptation to the host environment.

  6. Effect of acid whey and freeze-dried cranberries on lipid oxidation and fatty acid composition of nitrite-/nitrate-free fermented sausage made from deer meat

    Directory of Open Access Journals (Sweden)

    Małgorzata Karwowska

    2017-01-01

    Full Text Available Objective This study evaluated the effect of acid whey and freeze-dried cranberries on the physicochemical characteristics, lipid oxidation and fatty acid composition of nitrite-free fermented sausage made from deer meat and pork fat. Antioxidant interactions between acid whey and cranberry compounds were also explored. Methods Four formulations of fermented venison sausage were prepared: F1 (control, F2 (with 5% liquid acid whey, F3 (with 0.06% of freeze-dried cranberries, and F4 (with 5% liquid acid whey and 0.06% of freeze-dried cranberries. Each sample was analyzed for pH, water activity (aw, heme iron content, 2-thiobarbituric acid reactive substances (TBARS value and conjugated dienes at the end of the manufacturing process and at 30 and 90 days of refrigerated storage. Fatty acid composition was measured once at the end of the manufacturing process. Results At the end of ripening, all samples presented statistically different values for a pH range of 4.47 to pH 4.59. The sum of the unsaturated fatty acids was higher, while the conjugated diene and the TBARS values were lower in sausages with freeze-dried cranberries as compared to the control sausage. The highest content of heme iron (21.52 mg/kg at day 90 was found in the sausage formulation with the addition of freeze-dried cranberries, which suggests that the addition of cranberries stabilized the porphyrin ring of the heme molecule during storage and thereby reduced the release of iron. The use of liquid acid whey in combination with cranberries appears to not be justified in view of the oxidative stability of the obtained products. Conclusion The results suggest that the application of freeze-dried cranberries can lower the intensity of oxidative changes during the storage of nitrite-free fermented sausage made from deer meat.

  7. Serum-Free Cryopreservation of Five Mammalian Cell Lines in Either a Pelleted or Suspended State

    Directory of Open Access Journals (Sweden)

    Corsini Joe

    2004-01-01

    Full Text Available Herein we have explored two practical aspects of cryopreserving cultured mammalian cells during routine laboratory maintenance. First, we have examined the possibility of using a serum-free, hence more affordable, cryopreservative. Using five mammalian lines (Crandell Feline Kidney, MCF7, A72, WI 38 and NB324K, we found that the serum-free alternative preserves nearly as efficiently as the serum-containing preservatives. Second, we compared cryostorage of those cells in suspended versus a pellet form using both aforementioned cryopreservatives. Under our conditions, cells were in general recovered equally well in a suspended versus a pellet form.

  8. Development of a serum-free co-culture of human intestinal epithelium cell-lines (Caco-2/HT29-5M21)

    Science.gov (United States)

    Nollevaux, Géraldine; Devillé, Christelle; El Moualij, Benaïssa; Zorzi, Willy; Deloyer, Patricia; Schneider, Yves-Jacques; Peulen, Olivier; Dandrifosse, Guy

    2006-01-01

    Background The absorptive and goblet cells are the main cellular types encountered in the intestine epithelium. The cell lineage Caco-2 is a model commonly used to reproduce the features of the bowel epithelium. However, there is a strong debate regarding the value of Caco-2 cell culture to mimick in vivo situation. Indeed, some authors report in Caco-2 a low paracellular permeability and an ease of access of highly diffusible small molecules to the microvilli, due to an almost complete lack of mucus. The HT29-5M21 intestinal cell lineage is a mucin-secreting cellular population. A co-culture system carried out in a serum-free medium and comprising both Caco-2 and HT29-5M21 cells was developed. The systematic use of a co-culture system requires the characterization of the monolayer under a given experimental procedure. Results In this study, we investigated the activity and localization of the alkaline phosphatase and the expression of IAP and MUC5AC genes to determine a correlation between these markers and the cellular composition of a differentiated monolayer obtained from a mixture of Caco-2 and HT29-5M21 cells. We observed that the culture conditions used (serum-free medium) did not change the phenotype of each cell type, and produced a reproducible model. The alkaline phosphatase expression characterizing Caco-2 cells was influenced by the presence of HT29-5M21 cells. Conclusion The culture formed by 75% Caco-2 and 25% HT29-5M21 produce a monolayer containing the two main cell types of human intestinal epithelium and characterized by a reduced permeability to macromolecules. PMID:16670004

  9. Alpha-ketoglutarate enhances freeze-thaw tolerance and prevents carbohydrate-induced cell death of the yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Bayliak, Maria M; Hrynkiv, Olha V; Knyhynytska, Roksolana V; Lushchak, Volodymyr I

    2018-01-01

    Stress resistance and fermentative capability are important quality characteristics of baker's yeast. In the present study, we examined protective effects of exogenous alpha-ketoglutarate (AKG), an intermediate of the tricarboxylic acid cycle and amino acid metabolism, against freeze-thaw and carbohydrate-induced stresses in the yeast Saccharomyces cerevisiae. Growth on AKG-supplemented medium prevented a loss of viability and improved fermentative capacity of yeast cells after freeze-thaw treatment. The cells grown in the presence of AKG had higher levels of amino acids (e.g., proline), higher metabolic activity and total antioxidant capacity, and higher activities of catalase, NADP-dependent glutamate dehydrogenase and glutamine synthase compared to control ones. Both synthesis of amino acids and enhancement of antioxidant system capacity could be involved in AKG-improved freeze-thaw tolerance in S. cerevisiae. Cell viability dramatically decreased under incubation of stationary-phase yeast cells in 2% glucose or fructose solutions (in the absence of the other nutrients) as compared with incubation in distilled water or in 10 mM AKG solution. The decrease in cell viability was accompanied by acidification of the medium, and decrease in cellular respiration, aconitase activity, and levels of total protein and free amino acids. The supplementation with 10 mM AKG effectively prevented carbohydrate-induced yeast death. Protective mechanisms of AKG could be associated with the intensification of respiration and prevention of decreasing protein level as well as with direct antioxidant AKG action.

  10. Serum-free microcarrier based production of replication deficient Influenza vaccine candidate virus lacking NS1 using Vero cells

    Directory of Open Access Journals (Sweden)

    Yan Mylene L

    2011-08-01

    first time the production of Influenza viruses using Vero cells in commercially available animal-component free, serum-free medium. This work can be used as a basis for efficient production of attenuated as well as wild type Influenza virus for research and vaccine production.

  11. In which patients should serum creatinine be measured before iodinated contrast medium administration?

    International Nuclear Information System (INIS)

    Thomsen, Henrik S.

    2005-01-01

    Routine measurement of serum creatinine before injection of intravascular iodinated contrast material in all patients would be cumbersome and have an associated cost. There is doubt about whether serum creatinine should be measured routinely in all patients or selectively. The Contrast Media Safety Committee of the European Society of Urogenital Radiology decided to review the literature and draw up guidelines on this important practical issue. A literature search was carried out and summarized in a report. Based on the available information and discussions amongst the members of the Committee, guidelines were produced. The report and guidelines were discussed at the 11th European Symposium on Urogenital Radiology in Santiago de Compostela, Spain. The practice for identifying patients at risk of contrast medium induced nephropathy varies considerably from one institution to another. Patients at risk constitute only a small percentage of all cases referred for contrast enhanced imaging examination. However, it is important to identify them and take the necessary precautions. Recent serum creatinine level should be available in patients with an increased probability of a raised serum creatinine level (renal disease, renal surgery, proteinuria, diabetes mellitus, hypertension, gout, current intake of nephrotoxic drugs). A simple guideline has been produced. (orig.)

  12. In which patients should serum creatinine be measured before iodinated contrast medium administration?

    Energy Technology Data Exchange (ETDEWEB)

    Thomsen, Henrik S. [Copenhagen University Hospital, Herlev (Denmark). Department of Diagnostic Radiology; Morcos, Sameh K. [Northern General Hospital, Department of Diagnostic Imaging, Sheffield Teaching Hospitals NHS Trust, Sheffield (United Kingdom)

    2005-04-01

    Routine measurement of serum creatinine before injection of intravascular iodinated contrast material in all patients would be cumbersome and have an associated cost. There is doubt about whether serum creatinine should be measured routinely in all patients or selectively. The Contrast Media Safety Committee of the European Society of Urogenital Radiology decided to review the literature and draw up guidelines on this important practical issue. A literature search was carried out and summarized in a report. Based on the available information and discussions amongst the members of the Committee, guidelines were produced. The report and guidelines were discussed at the 11th European Symposium on Urogenital Radiology in Santiago de Compostela, Spain. The practice for identifying patients at risk of contrast medium induced nephropathy varies considerably from one institution to another. Patients at risk constitute only a small percentage of all cases referred for contrast enhanced imaging examination. However, it is important to identify them and take the necessary precautions. Recent serum creatinine level should be available in patients with an increased probability of a raised serum creatinine level (renal disease, renal surgery, proteinuria, diabetes mellitus, hypertension, gout, current intake of nephrotoxic drugs). A simple guideline has been produced. (orig.)

  13. A novel liquid medium for the efficient growth of the salmonid pathogen Piscirickettsia salmonis and optimization of culture conditions.

    Directory of Open Access Journals (Sweden)

    Mirtha Henríquez

    Full Text Available Piscirickettsia salmonis is the bacterium that causes Piscirickettsiosis, a systemic disease of salmonid fish responsible for significant economic losses within the aquaculture industry worldwide. The growth of the bacterium for vaccine formulation has been traditionally accomplished by infecting eukaryotic cell lines, a process that involves high production costs and is time-consuming. Recent research has demonstrated that it is possible to culture pure P. salmonis in a blood containing (cell-free medium. In the present work we demonstrate the growth of P. salmonis in a liquid medium free from blood and serum components, thus establishing a novel and simplified bacteriological medium. Additionally, the new media reported provides improved growth conditions for P. salmonis, where biomass concentrations of approximately 800 mg cell dry weight L(-1 were obtained, about eight times higher than those reported for the blood containing medium. A 2- level full factorial design was employed to evaluate the significance of the main medium components on cell growth and an optimal temperature range of 23-27°C was determined for the microorganism to grow in the novel liquid media. Therefore, these results represent a breakthrough regarding P. salmonis research in order to optimize pure P. salmonis growth in liquid blood and serum free medium.

  14. Utility of serum periostin and free IgE levels in evaluating responsiveness to omalizumab in patients with severe asthma.

    Science.gov (United States)

    Tajiri, T; Matsumoto, H; Gon, Y; Ito, R; Hashimoto, S; Izuhara, K; Suzukawa, M; Ohta, K; Ono, J; Ohta, S; Ito, I; Oguma, T; Inoue, H; Iwata, T; Kanemitsu, Y; Nagasaki, T; Niimi, A; Mishima, M

    2016-10-01

    Omalizumab, a humanized anti-IgE monoclonal antibody, has demonstrated efficacy in patients with severe allergic asthma. However, treatment responses vary widely among individuals. Despite a lack of data, free serum IgE levels following omalizumab treatment have been proposed as a marker of treatment responsiveness. In this prospective, observational study, we assessed the utility of biomarkers of type 2 inflammation in predicting omalizumab treatment responses, as determined by the absence of asthma exacerbation during the first year of treatment. Free serum IgE levels were monitored for 2 years to examine their association with baseline biomarker levels and the number of exacerbations. We enrolled thirty patients who had been treated with omalizumab for at least 1 year, of whom 27 were treated for 2 years. Baseline serum periostin levels and blood eosinophil counts were significantly higher in patients without exacerbations during the first year of treatment than in patients with exacerbations. Baseline serum periostin levels, but not eosinophil counts, were negatively associated with free serum IgE levels after 16 or 32 weeks of treatment. Reduced free serum IgE levels during treatment from those at baseline were associated with reduced exacerbation numbers at 2 years. In 14 patients who continued to have exacerbations during the first year of treatment, exacerbation numbers gradually and significantly decreased over the 2-year study period, with concurrent significant reductions in free serum IgE levels. Baseline serum periostin levels and serum free IgE levels during treatment follow-up may be useful in evaluating responses to omalizumab treatment. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  15. Role of hemolysis in potassium release by iodinated contrast medium

    Energy Technology Data Exchange (ETDEWEB)

    Hayakawa, K.; Nakamura, T.; Shimizu, Y. [Department of Radiology, Kyoto City Hospital (Japan)

    1999-09-01

    It has been demonstrated that an iodinated contrast medium (CM) causes release of potassium into blood vessel lumina, resulting in an increase in serum potassium. The purpose of the present study was to assess whether this potassium release is due to hemolysis. Fresh human blood was mixed in vitro with CM at a ratio of 10:2. Potassium release rates were determined, and serum haptoglobin and free hemoglobin were measured after 30 min of exposure to CM. To compare the potassium release curve between CM exposure and true hemolysis induced by distilled water, fresh human blood was also mixed with distilled water. The level of serum haptoglobin decreased due to hemodilution. Changes in haptoglobin were not correlated with potassium release rates. The serum free hemoglobin level did not increase significantly, and there was no correlation between changes in the free hemoglobin level and the rate of potassium release. Hemolysis caused by water occurred instantaneously, whereas potassium release caused by CM was a slow response, which was linearly correlated with exposure time. Potassium release from blood cannot be explained by hemolysis. (orig.) With 4 figs., 4 tabs., 3 refs.

  16. Freeze concentration of lime juice

    Directory of Open Access Journals (Sweden)

    Ampawan Tansakul

    2008-11-01

    Full Text Available The main objective of this research was to study the effects of processing conditions, i.e. cooling medium temperature (-6, -12 and -18C and scraper blade rotational speed (50, 100 and 150 rpm on the freeze concentration of lime juice. The initial soluble solid content of lime juice was 7.6 Brix. Results showed that soluble solid content of lime juice increased as cooling medium temperature decreased while scraper blade rotational speed increased. It was also found that the processing condition with -18˚C cooling medium temperature and 150 rpm rotational speed of the scraper blade was the best among all studied conditions, although the loss of the soluble solids with ice crystals during ice separation was relatively high at 35%.

  17. Evolution of triiodothyronine nuclear binding sites in hypothalamic serum-free cultures: evidence for their presence in neurons and astrocytes

    International Nuclear Information System (INIS)

    Puymirat, J.; Faivre-Bauman, A.

    1986-01-01

    ( 125 I)Triiodothyronine (T 3 ) nuclear binding was studied in hypothalamic cultures from fetal mouse grown in serum-free medium. In enriched neuronal cultures, the apparent dissociation constant of the binding does not change with time in vitro (7 x 10 -11 M), but the maximum binding capacity (MBC) doubles between day 7 and day 14 in vitro. We show here for the first time that homologous astrocyte cell cultures, devoid of neurons as checked by tetanus toxin binding, also display T 3 nuclear binding, with the same affinity as neuronal cultures. However, their MBC is 3 times lower than that of neurons after a week in vitro, and increases more quickly thereafter (Author)

  18. Accelerated storage testing of freeze-dried Pseudomonas ...

    African Journals Online (AJOL)

    Erika-Chris

    Full Length Research Paper ... was possible to compare rate constants of survival for the freeze-dried P. fluorescens ... studying and predicting the survival loss rate of the ... Erlenmeyer flask containing 3000 ml King B medium. ... The strain was grown in 20 L bioreactor (Biolafite) containing 15 L .... fermented banana media.

  19. Serum and plasma for total and free anticonvulsant drug analyses: effects on EMIT assays and ultrafiltration devices.

    Science.gov (United States)

    Godolphin, W; Trepanier, J; Farrell, K

    1983-01-01

    The suitability of serum and plasma anticoagulated with heparin, EDTA, citrate, or oxalate was assessed for analysis of free and total phenytoin, carbamazepine, and valproic acid. The free fraction was isolated by ultrafiltration through FreeLevel devices (Syva, Palo Alto, CA). Serum, heparin, and EDTA plasma were satisfactory for both free and total phenytoin and carbamazepine. EDTA could not be used for EMIT (Syva) analysis of valproate. Citrate and, to a lesser degree, oxalate cause a significant negative interference in the concentration of these three drugs as measured both by EMIT and gas-liquid chromatography.

  20. Embryogenesis-promoting factors in rat serum.

    Science.gov (United States)

    Katoh, M; Kimura, R; Shoji, R

    1998-06-15

    Regarding whole rat embryo cultures in vitro, rat serum as a culture medium is known to support the normal growth of rat embryos in the organogenesis phase. The purpose of the present study was to isolate the embryogenesis-promoting factors from rat serum as a first step in the development of a defined serum-free medium for a whole embryo culture system. Pooled rat serum after heat inactivation was fractionated into three major peaks (frA, containing a region of void volume, frB, and frC) by gel filtration. The 9.5-day rat embryos that were cultivated for 48 hr in essential salt medium containing frB (with a molecular size range of 100-500 kDa) revealed normal growth. Three proteins (27 kDa, 76 kDa, and 190 kDa) that had the embryogenesis-promoting effects were isolated from 3-hr delayed centrifuged rat serum by the ion exchange chromatography. The 76-kDa protein was found to be rat transferrin by immunoblotting. The 27-kDa protein was identified as apo-AI (the major apoprotein of high-density lipoprotein) by immunoblotting. High-density lipoprotein obtained from pooled rat serum by a NaBr density gradient ultracentrifugation was found to have a positive effect on embryogenesis. The 10-kDa protein was also identified as alpha 1-inhibitor 3 by immunoblotting. In addition, the embryogenesis-promoting effect of the fraction containing 27-kDa and 190-kDa proteins declined within a short period of storage at -20 degrees C. This decrease was countered by supplementing its fraction (D-2) with albumin isolated from rat serum. These results in the present study suggest that transferrin, high-density lipoprotein, and alpha 1-inhibitor 3 in rat serum may be embryogenesis-promoting factors, and that albumin appeared to play a role in the embryogenesis of rat embryos in whole embryo cultures.

  1. Newly-derived neuroblastoma cell lines propagated in serum-free media recapitulate the genotype and phenotype of primary neuroblastoma tumours.

    Science.gov (United States)

    Bate-Eya, Laurel T; Ebus, Marli E; Koster, Jan; den Hartog, Ilona J M; Zwijnenburg, Danny A; Schild, Linda; van der Ploeg, Ida; Dolman, M Emmy M; Caron, Huib N; Versteeg, Rogier; Molenaar, Jan J

    2014-02-01

    Recently protocols have been devised for the culturing of cell lines from fresh tumours under serum-free conditions in defined neural stem cell medium. These cells, frequently called tumour initiating cells (TICs) closely retained characteristics of the tumours of origin. We report the isolation of eight newly-derived neuroblastoma TICs from six primary neuroblastoma tumours and two bone marrow metastases. The primary tumours from which these TICs were generated have previously been fully typed by whole genome sequencing (WGS). Array comparative genomic hybridisation (aCGH) analysis showed that TIC lines retained essential characteristics of the primary tumours and exhibited typical neuroblastoma chromosomal aberrations such as MYCN amplification, gain of chromosome 17q and deletion of 1p36. Protein analysis showed expression for neuroblastoma markers MYCN, NCAM, CHGA, DBH and TH while haematopoietic markers CD19 and CD11b were absent. We analysed the growth characteristics and confirmed tumour-forming potential using sphere-forming assays, subcutaneous and orthotopic injection of these cells into immune-compromised mice. Affymetrix mRNA expression profiling of TIC line xenografts showed an expression pattern more closely mimicking primary tumours compared to xenografts from classical cell lines. This establishes that these neuroblastoma TICs cultured under serum-free conditions are relevant and useful neuroblastoma tumour models. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. Development of a serum-free co-culture of human intestinal epithelium cell-lines (Caco-2/HT29-5M21

    Directory of Open Access Journals (Sweden)

    Schneider Yves-Jacques

    2006-05-01

    Full Text Available Abstract Background The absorptive and goblet cells are the main cellular types encountered in the intestine epithelium. The cell lineage Caco-2 is a model commonly used to reproduce the features of the bowel epithelium. However, there is a strong debate regarding the value of Caco-2 cell culture to mimick in vivo situation. Indeed, some authors report in Caco-2 a low paracellular permeability and an ease of access of highly diffusible small molecules to the microvilli, due to an almost complete lack of mucus. The HT29-5M21 intestinal cell lineage is a mucin-secreting cellular population. A co-culture system carried out in a serum-free medium and comprising both Caco-2 and HT29-5M21 cells was developed. The systematic use of a co-culture system requires the characterization of the monolayer under a given experimental procedure. Results In this study, we investigated the activity and localization of the alkaline phosphatase and the expression of IAP and MUC5AC genes to determine a correlation between these markers and the cellular composition of a differentiated monolayer obtained from a mixture of Caco-2 and HT29-5M21 cells. We observed that the culture conditions used (serum-free medium did not change the phenotype of each cell type, and produced a reproducible model. The alkaline phosphatase expression characterizing Caco-2 cells was influenced by the presence of HT29-5M21 cells. Conclusion The culture formed by 75% Caco-2 and 25% HT29-5M21 produce a monolayer containing the two main cell types of human intestinal epithelium and characterized by a reduced permeability to macromolecules.

  3. Serum-free keloid fibroblast cell culture: an in vitro model for the study of aberrant wound healing.

    Science.gov (United States)

    Koch, R J; Goode, R L; Simpson, G T

    1997-04-01

    The purpose of this study was to develop an in vitro serum-free keloid fibroblast model. Keloid formation remains a problem for every surgeon. Prior evaluations of fibroblast characteristics in vitro, especially those of growth factor measurement, have been confounded by the presence of serum-containing tissue culture media. The serum itself contains growth factors, yet has been a "necessary evil" to sustain cell growth. The design of this study is laboratory-based and uses keloid fibroblasts obtained from five patients undergoing facial (ear lobule) keloid removal in a university-affiliated clinic. Keloid fibroblasts were established in primary cell culture and then propagated in a serum-free environment. The main outcome measures included sustained keloid fibroblast growth and viability, which was comparable to serum-based models. The keloid fibroblast cell cultures exhibited logarithmic growth, sustained a high cellular viability, maintained a monolayer, and displayed contact inhibition. Demonstrating model consistency, there was no statistically significant difference between the mean cell counts of the five keloid fibroblast cell lines at each experimental time point. The in vitro growth of keloid fibroblasts in a serum-free model has not been done previous to this study. The results of this study indicate that the proliferative characteristics described are comparable to those of serum-based models. The described model will facilitate the evaluation of potential wound healing modulators, and cellular effects and collagen modifications of laser resurfacing techniques, and may serve as a harvest source for contaminant-free fibroblast autoimplants. Perhaps its greatest utility will be in the evaluation of endogenous and exogenous growth factors.

  4. Determination of free amino acids of porcine serum responsible for ...

    African Journals Online (AJOL)

    The 1H NMR spectra of serum metabolites at 600 MHz showed that free amino acids such as alanine, leucine, phenylalanine, and valine were qualitatively higher in the HpHG than in the LpHG. The relative abundance of three amino acids was quantitatively verified by HPLC: Phenylalanine and valine (P<0.01) and leucine ...

  5. Serum Levels of Follistatin Are Positively Associated With Serum-Free Thyroxine Levels in Patients With Hyperthyroidism or Euthyroidism

    Science.gov (United States)

    Tseng, Fen-Yu; Chen, Yen-Ting; Chi, Yu-Chao; Chen, Pei-Lung; Yang, Wei-Shiung

    2016-01-01

    Abstract Follistatin is a glycoprotein with various biologic functions that plays a role in adipocyte differentiation, muscle stimulation, anti-inflammation, and energy homeostasis. Thyroid hormones influence energy expenditure, glucose, and lipid metabolism. The association between serum follistatin level and thyroid function statuses has seldom been evaluated. The objectives of this study were to compare serum follistatin concentrations in different thyroid function statuses and to evaluate the associations between serum follistatin and free thyroxine (fT4) levels. In this study, 30 patients with hyperthyroidism (HY group) and 30 euthyroid individuals (EU group) were recruited. The patients of HY group were treated with antithyroid regimens as clinically indicated, whereas no medication was given to EU group. The demographic and anthropometric characteristics, biochemical data, serum levels of follistatin, and thyroid function of both groups at baseline and at the 6th month were compared. Data of all patients were pooled for the analysis of the associations between the levels of follistatin and fT4. At baseline, the HY group had significantly higher serum follistatin levels than the EU group (median [Q1, Q3]: 1.81 [1.33, 2.78] vs 1.13 [0.39, 1.45] ng/mL, P hyperthyroidism had higher serum follistatin levels, which decreased after receiving antithyroid treatment. In addition, the serum follistatin concentrations were positively associated with serum fT4 levels in patients with hyperthyroidism or euthyroidism. PMID:26844494

  6. Ultrasound assisted immersion freezing of broccoli (Brassica oleracea L. var. botrytis L.).

    Science.gov (United States)

    Xin, Ying; Zhang, Min; Adhikari, Benu

    2014-09-01

    The aim of this study was to research the ultrasound-assisted freezing (UAF) of broccoli. CaCl2 solution was used as freezing medium. The comparative advantage of using UAF over normal freezing on the freezing time, cell-wall bound calcium to total calcium ratio, textural properties, color, drip loss and L-ascorbic acid contents was evaluated. The application of UAF at selected acoustic intensity with a range of 0.250-0.412 W/cm(2) decreased the freezing time and the loss of cell-wall bound calcium content. Compared to normal freezing, the values of textural properties, color, L-ascorbic acid content were better preserved and the drip loss was significantly minimized by the application of UAF. However, when outside that range of acoustic intensity, the quality of the ultrasound-assisted frozen broccoli was inferior compared to that of the normally frozen samples. Selected the appropriate acoustic intensity was very important for the application of UAF. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Serum total and free carnitine levels in children with asthma.

    Science.gov (United States)

    Asilsoy, Suna; Bekem, Ozlem; Karaman, Ozkan; Uzuner, Nevin; Kavukçu, Salih

    2009-02-01

    Serum carnitine is decreased in recurrent pulmonary infections. We aimed to evaluate serum carnitine levels in asthmatic children. Study group consisted of children with stable asthma and those with acute asthma attacks, while control group included healthy children. Attack severity was determined by the pulmonary score system. Total and free carnitine levels were studied in one blood sample from the control group and stable asthmatics and in two samples from children with acute asthma exacerbation during and after the attack. All the 40 patients in the study group had moderate asthma including 30 with acute attack (13 mild and 17 moderate) and 10 with stable asthma. Carnitine levels were significantly lower in acute attack asthmatics than in the stable asthmatics and controls, while there was no significant difference between the latter two groups. Carnitine levels were not different between asthmatics with mild and moderate attack, and were similar during and after an acute attack. Serum carnitine levels decrease in children with moderate asthma during exacerbation of asthma and shortly thereafter. Further studies are needed to evaluate the effect of carnitine treatment on serum carnitine level.

  8. Determination of free amino acids and 18 elements in freeze-dried strawberry and blueberry fruit using an Amino Acid Analyzer and ICP-MS with micro-wave digestion.

    Science.gov (United States)

    Zhang, Hua; Wang, Zhen-Yu; Yang, Xin; Zhao, Hai-Tian; Zhang, Ying-Chun; Dong, Ai-Jun; Jing, Jing; Wang, Jing

    2014-03-15

    The objective of this study was to investigate the level of 18 trace elements of two freeze-dried samples from the Blueberry (Vaccinium corymbosum) and the Strawberry (Fragaria × Ananassa). The total free amino acid composition in the blueberry and strawberry was determined by an Amino Acid Analyzer. Eleven free amino acids were found in both berries. The trace elements in each dried fruit sample were determined by ICP-MS with microwave digestion. The linearity range of the standard curves was 0-1250.0 μg L(-1) (Mg, P, K, Ca),while in all cases, except for B, Na, Al, Cr, Mn, Fe, Ni, Cu, Zn, Se, Cd, Pb, Ge and As, which was 125.0 μg mL(-1), all related coefficients were above 0.9999; recovery was in the range of 79.0-106.8%. Minor concentrations of nutritional elements were found in each freeze-dried berry. In sum, the toxic trace element analysis found the content of toxic trace elements in each freeze-dried berry sample was safe for human consumption and that the overall quality of the blueberry surpassed that of the strawberry. The results certify that the two freeze-dried berries have potential for human consumption in value-added products and have a certain theoretical and practical significance. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

  9. The influence of a cryoprotective medium containing glycerol on the lyophilization of lactic acid bacteria (NOTE

    Directory of Open Access Journals (Sweden)

    JOSIP BARAS

    2001-07-01

    Full Text Available The aims of liophilization (freeze-drying of lactic acid bacteria are to preserve pure cultures or to prepare starters for the dairy industry. In both cases, the choice of the cryoprotectant is very important. In this work, samples of Bifidobacterium breve A71 and Bifidobacterium bifidum BbTD were freeze-dried in a new cryoprotective medium containing lactose, gelatine and glycerol (medium B. The reference medium contained saccharose, gelatine and skim milk (medium A. Before liophilization, the eutectic points of both media were determined, because the products must be cooled to a temperature below its freezing point. The success of the cryoprotectants was estimated in terms of the number of surviving organisms after lyophilization. Bifidobacterium breve A71 and Bifidobacterium bifidum BbTD freeze-dried in media A and B showed high survival rates. Bifidobacterium breve A71 showed a greater percentage survival in combination with medium B than with medium A. These results could be utilized in the manufacture of Bifidobacterium breve A71 as a starter in the diary industry because it is a human isolate which, except for acidification, has probiotic activity.

  10. Tumour-cytolytic human monocyte-derived macrophages: a simple and efficient method for the generation and long-term cultivation as non-adherent cells in a serum-free medium.

    Science.gov (United States)

    Streck, R J; Hurley, E L; Epstein, D A; Pauly, J L

    1992-01-01

    We report a simple and efficient culture procedure for the generation of tumour-cytolytic human monocyte-derived macrophages (MAC). In this method, normal human peripheral blood mononuclear cells, isolated using a conventional Ficoll-Hypaque density gradient procedure, are cultured as a heterogenous leukocyte population in Teflon or other hydrophobic cultureware, in a commercially available serum-free culture medium (M-SFM) that has been formulated specifically for the cultivation and ex vivo stimulation of human monocytes and MAC, and in the absence of exogenous mitogens, antigens, cytokines or other stimulants. This procedure features a negative-selection technique that takes advantage of the differential survival of blood leukocytes. Using the prescribed in vitro conditions, lymphocytes survived relatively poorly, whereas monocytes differentiated in the absence of exogenous stimulants into mature tumour-cytolytic MAC. The MAC were present as non-adherent, single cells that expressed good viability (greater than 95%) for a prolonged period (greater than 60 days). When compared to conventional procedures for generating MAC, the prescribed technique is thought to offer several important advantages in that it: (a) eliminates the tedious and cumbersome monocyte isolation procedures, thus providing a significant savings not only in time and money but also in eliminating repetitive cell manipulations that have often been associated with damage to monocyte morphology and/or function; (b) reduces the loss of monocyte subsets that are not recovered during specific isolation procedures; (c) facilitates harvesting a single cell, non-adherent suspension of immunocompetent MAC suitable for various examinations including analyses defining MAC morphology, cytochemistry, phenotype and function; and (d) eliminates variability and artifacts associated with different sera that are utilised frequently as medium supplements. The utility of the prescribed method is illustrated by the

  11. Non-covalent interaction between dietary stilbenoids and human serum albumin: Structure-affinity relationship, and its influence on the stability, free radical scavenging activity and cell uptake of stilbenoids.

    Science.gov (United States)

    Cao, Hui; Jia, Xueping; Shi, Jian; Xiao, Jianbo; Chen, Xiaoqing

    2016-07-01

    Dietary stilbenoids are associated with many benefits for human health, which depend on their bioavailability and bioaccessibility. The stilbenoid-human serum albumin (HSA) interactions are investigated to explore the structure-affinity relationship and influence on the stability, free radical scavenging activity and cell uptake of stilbenoids. The structure-affinity relationship of the stilbenoids-HSA interaction was found as: (1) the methoxylation enhanced the affinity, (2) an additional hydroxyl group increases the affinity and (3) the glycosylation significantly weakened the affinity. HSA obviously masked the free radical scavenging potential of stilbenoids. The stabilities of stilbenoids in different medium were determined as: HSA solution>human plasma>Dulbecco's modified Eagle's medium. It appears that the milk enhanced the cell uptake of stilbenoids with multi-hydroxyl groups and weakened the cell uptake of stilbenoids with methoxyl group on EA.hy 926 endothelial cells. The stilbenoids are hardly absorbed by human umbilical vein endothelial cells in the presence of milk. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Thermal processing of bone: in vitro response of mesenchymal cells to bone-conditioned medium.

    Science.gov (United States)

    Sawada, K; Caballé-Serrano, J; Schuldt Filho, G; Bosshardt, D D; Schaller, B; Buser, D; Gruber, R

    2015-08-01

    The autoclaving, pasteurization, and freezing of bone grafts to remove bacteria and viruses, and for preservation, respectively, is considered to alter biological properties during graft consolidation. Fresh bone grafts release paracrine-like signals that are considered to support tissue regeneration. However, the impact of the autoclaving, pasteurization, and freezing of bone grafts on paracrine signals remains unknown. Therefore, conditioned medium was prepared from porcine cortical bone chips that had undergone thermal processing. The biological properties of the bone-conditioned medium were assessed by examining the changes in expression of target genes in oral fibroblasts. The data showed that conditioned medium obtained from bone chips that had undergone pasteurization and freezing changed the expression of adrenomedullin, pentraxin 3, BTB/POZ domain-containing protein 11, interleukin 11, NADPH oxidase 4, and proteoglycan 4 by at least five-fold in oral fibroblasts. Bone-conditioned medium obtained from autoclaved bone chips, however, failed to change the expression of the respective genes. Also, when bone-conditioned medium was prepared from fresh bone chips, autoclaving blocked the capacity of bone-conditioned medium to modulate gene expression. These in vitro results suggest that pasteurization and freezing of bone grafts preserve the release of biologically active paracrine signals, but autoclaving does not. Copyright © 2015 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  13. Serum free-thyroxine modifications with age and in normal pregnancy

    International Nuclear Information System (INIS)

    Degrossi, O.J.; Altschuler, N.; Watanabe, T.; Pinkas, M.; Damilano, S.; Garcia del Rio, H.

    1982-01-01

    The possibility of using radioimmunoassay techniques (RIA) in the assessment of circulating thyroid hormones, total thyroxine (T4) and triiodothyronine (T3) has particularly increases the diagnosis of thyroid diseases. Thyroidal hormones circulate bound to proteins; therefore, variations in the transport capacity of the latter will produce important modifications in the T4 and T3 figures. Only small fractions, less than 0.05% for T4 and than 0.5% for T3, circulate in the free form and are considered as metabolically active forms of both hormones. In order to attain a correct clinical valuation, the rates of the free fractions as well as the total rates of these hormones must be known. Recently, these studies of free hormones are carried out by means of RIA, with the consequent advantages. The variations in serum free thyroxine (FT4) under certain physiogical conditions, such as for different ages and during pregnancy, were particularly studied

  14. Generalized enthalpy model of a high-pressure shift freezing process

    KAUST Repository

    Smith, N. A. S.

    2012-05-02

    High-pressure freezing processes are a novel emerging technology in food processing, offering significant improvements to the quality of frozen foods. To be able to simulate plateau times and thermal history under different conditions, in this work, we present a generalized enthalpy model of the high-pressure shift freezing process. The model includes the effects of pressure on conservation of enthalpy and incorporates the freezing point depression of non-dilute food samples. In addition, the significant heat-transfer effects of convection in the pressurizing medium are accounted for by solving the two-dimensional Navier-Stokes equations. We run the model for several numerical tests where the food sample is agar gel, and find good agreement with experimental data from the literature. © 2012 The Royal Society.

  15. Increased serum free tryptophan in patients with diarrhea-predominant irritable bowel syndrome.

    Science.gov (United States)

    Christmas, David M; Badawy, Abdulla A-B; Hince, Dana; Davies, Simon J C; Probert, Christopher; Creed, Tom; Smithson, John; Afzal, Muhammad; Nutt, David J; Potokar, John P

    2010-10-01

    Irregularities of serotonin function in irritable bowel syndrome (IBS) may be due to changes in the metabolism of the serotonin precursor l-tryptophan. Dietary alteration of tryptophan intake may impact upon the mood and bowel symptoms of IBS. We hypothesized that diarrhea-predominant irritable bowel syndrome (d-IBS) patients would exhibit an increase in plasma tryptophan due to alterations in tryptophan metabolism. We also hypothesized that a diet low in tryptophan would reverse this change and reduce symptoms. Thirteen patients with d-IBS had fasting serum free and total tryptophan, large neutral amino acids, and 6 kynurenine metabolites measured before and after 2 weeks of a strict dairy-free diet. Baseline tryptophan parameters were compared with an age- and sex-matched control group. Changes in the specific tryptophan parameters before and after dairy-free diet were correlated with symptoms of IBS and mood. Compared with the control group, d-IBS patients at baseline exhibited significantly higher free serum tryptophan (10.5 ± 4.35 vs 4.75 ± 2.43 μmol/L [means ± standard deviation], P = .006) and significantly lower tryptophan dioxygenase and total tryptophan oxidation as measured by the kynurenine to free tryptophan and total kynurenines to free tryptophan ratios (23.37 ± 10.12 vs 55.33 ± 16.02, P < .001 and 49.34 ± 17.84 vs 258.46 ± 98.67, P < .001, respectively). Dairy-free diet did not modulate metabolites of the kynurenine pathway or symptoms. Tryptophan metabolism along the kynurenine pathway is inhibited in d-IBS, and a dairy-free diet does not alter this. Our findings are consistent with possible enhanced serotonin activity in d-IBS. Copyright © 2010 Elsevier Inc. All rights reserved.

  16. Trypanosoma brucei gambiense: HMI-9 medium containing methylcellulose and human serum supports the continuous axenic in vitro propagation of the bloodstream form.

    Science.gov (United States)

    Van Reet, N; Pyana, P P; Deborggraeve, S; Büscher, P; Claes, F

    2011-07-01

    Trypanosoma brucei (T.b.) gambiense causes the chronic form of human African trypanosomiasis or sleeping sickness. One of the major problems with studying T.b. gambiense is the difficulty to isolate it from its original host and the difficult adaptation to in vivo and in vitro mass propagation. The objective of this study was to evaluate if an established method for axenic culture of pleomorphic bloodstream form T.b. brucei strains, based on methylcellulose containing HMI-9 medium, also facilitated the continuous in vitro propagation of other bloodstream form Trypanozoon strains, in particular of T.b. gambiense. Bloodstream form trypanosomes from one T.b. brucei, two T.b. rhodesiense, one T. evansi and seven T.b. gambiense strains were isolated from mouse blood and each was concurrently cultivated in liquid and methylcellulose-containing HMI-9 based medium, either with or without additional human serum supplementation, for over 10 consecutive sub passages. Although HMI-9 based medium supplemented with 1.1% (w/v) methylcellulose supported the continuous cultivation of all non-gambiense strains better than liquid media could, the in vitro cultivation of all gambiense strains was only achieved in HMI-9 based medium containing 1.1% (w/v) methylcellulose, 15% (v/v) fetal calf serum and 5% (v/v) heat-inactivated human serum. Copyright © 2011 Elsevier Inc. All rights reserved.

  17. A solid phase radioimmunoassay for free triiodothyronine in serum: assay development and validation

    International Nuclear Information System (INIS)

    Liu Fangyan; Zhou Meiying; Yin Linxiang; Yang Jianzhong; Hua Chenglin

    1999-01-01

    A solid phase radioimmunoassay for free triiodothyronine in serum was developed based on double-antibody coated tubes. The method was turned out to be reliable with good reproducibility, higher sensitivity and easy performance. The measurable range of FT 3 in serum was 1.2 to 38 pmol/L. The mean coefficients of variation within and between assays were 1.79%∼3.18% and 4.72%∼9.31%, respectively. The FT 3 concentrations in euthyroid serum as determined by this method were 2.8 to 7.8 pmol/L. The FT 3 values determined by this new method correlated well with those measured by a commercial radioimmunoassay (r = 0.853)

  18. Transition from natural-convection-controlled freezing to conduction-controlled freezing

    International Nuclear Information System (INIS)

    Sparrow, E.M.; Ramsey, J.W.; Harris, J.S.

    1981-01-01

    Experiments were performed to study the transition between freezing controlled by natural convection in the liquid adjacent to a freezing interface and freezing controlled by heat conduction in the solidified material. The freezing took place on a cooled vertical tube immersed in an initially superheated liquid contained in an adiabatic-walled vessel. At early and intermediate times, temperature differences throughout the liquid induce a vigorous natural convection motion which retards freezing, but the temperature differences diminish with time and natural convection ebbs. At large times, the freezing rate is fully controlled by heat conduction in the solidified material. The frozen specimens for short and intermediate freezing times are smooth-surfaced and tapered, while those for large times are straight-sided and have surfaces that are overlaid with a thicket of large discrete crystals. These characteristics correspond respectively to those of natural-convection- controlled freezing and conduction-controlled freezing. At early times, the measured mass of the frozen material is identical to that for natural-convection-controlled freezing and conduction-controlled freezing. At early times, the measured mass of the frozen material is identical to that for natural-convection-controlled freezing. At later times, the frozen mass tends to approach that for conduction-controlled freezing, but a residual deficit remains

  19. Neuronal medium that supports basic synaptic functions and activity of human neurons in vitro.

    Science.gov (United States)

    Bardy, Cedric; van den Hurk, Mark; Eames, Tameji; Marchand, Cynthia; Hernandez, Ruben V; Kellogg, Mariko; Gorris, Mark; Galet, Ben; Palomares, Vanessa; Brown, Joshua; Bang, Anne G; Mertens, Jerome; Böhnke, Lena; Boyer, Leah; Simon, Suzanne; Gage, Fred H

    2015-05-19

    Human cell reprogramming technologies offer access to live human neurons from patients and provide a new alternative for modeling neurological disorders in vitro. Neural electrical activity is the essence of nervous system function in vivo. Therefore, we examined neuronal activity in media widely used to culture neurons. We found that classic basal media, as well as serum, impair action potential generation and synaptic communication. To overcome this problem, we designed a new neuronal medium (BrainPhys basal + serum-free supplements) in which we adjusted the concentrations of inorganic salts, neuroactive amino acids, and energetic substrates. We then tested that this medium adequately supports neuronal activity and survival of human neurons in culture. Long-term exposure to this physiological medium also improved the proportion of neurons that were synaptically active. The medium was designed to culture human neurons but also proved adequate for rodent neurons. The improvement in BrainPhys basal medium to support neurophysiological activity is an important step toward reducing the gap between brain physiological conditions in vivo and neuronal models in vitro.

  20. Changes in serum concentrations of total and free testosterone in young and middle-aged men

    International Nuclear Information System (INIS)

    Wang Guohong; Xu Ruiji; Zhang Zhongshu

    2011-01-01

    To determine changes in serum concentrations of total and free testosterone in young and middle-aged men.the healthy men (n=126) were divided 20-29 yr, 30-39 yr and 40-49 yr three groups,their serum levels of total testosterone (T), free testosterone (FT), luteinizing hormone (LH), follicle-stimulating hormone (FSH), sex hormone binding globulin (SHBG) and estradiol (E2) were detected. The results were statically analyzed. The results showed that the levels of serum T and FT was found significantly decreased in 30-39 yr group than in 20-29 yr group (15.06±13 nmol/L vs 20.41±86 nmol/L, P<0.01; 9.36±95 pg/L vs 11.48±88 pg/L, P<0.05; respectively). There were young trends that age-related decline in androgen levels. (authors)

  1. Analysis of relationship between tumor markers and quantification of free DNA in serum of lung cancer patients

    International Nuclear Information System (INIS)

    Yang Shunfang; Zhang Peiling; Cao Jie; Zeng Jun; Dong Qianggang

    2006-01-01

    To evaluate the diagnostic value and relationship between five tumor markers (CA19- 9,CA125,CYFRA21-1 ,CEA,NSE) and free DNA in serum for lung cancer detection and try to find a new and more efficient tumor marker, the amounts of CA19-9, CA125, CYFRA21-1, CEA, NSE were determined by RIA and free DNA was determined by the use of quantitative real time PCR amplification of the human epidermal growth factor receptor (EGFR) in 52 lung cancer patients and 8 cases of benign pulmonary disease and 10 healthy controls. The resulls showed that average concentration of free DNA in serum of lung cancer patients, benign pulmo- nary disease and healthy controls was 107.6ng/mL, 76.86ng/mL and 18.8ng/mL, respective- ly. The diagnostic sensitivity, specificity and accuracy of free DNA for lung cancer were 71. 2%, 50% and 68.3%, same as the diagnostic value of combined detection of five tumor markers. The sensitivity, specificity and accuracy of the five tumor markers and free DNA combinend detection for lung cancer were 94.2%, 25% and 85%, respectively. The free DNA in the serum of lung cancer patients may be a new and better tumor marker. (authors)

  2. PECULIARITIES OF WATER FREEZING IN CRYOPROTECTIVE MEDIUM IMPLEMENTED IN A MATRIX OF HYDROPHOBIC SILICA BULL SPERM

    Directory of Open Access Journals (Sweden)

    V. V. Turov

    2014-06-01

    Full Text Available The study of the process of melting water in lactose-glycerol-yolk kriomedium containing gametes bull, incorporated in the hydrophobic silica powder, which are adsorbed on the surface of fixed amounts of nonpolar hydrocarbon – n-decane was the aim of the work. The possibility of water polyassociates structuring with a solid surface of interfacial water and solubility of trifluoroethanoic acid in it have been studied. Thereat survival of the germ cell after contact with the surface was not analyzed. State of water in initial cryoprotective glycerol-lactose-yolk medium and hydrophobic nanosilica TS-100 containing n-decane additive adsorbed on its surface incorporated in a matrix was studied using low-temperature 1H-NMR spectroscopy method. It is shown that the solid matrix induces formation of 6–7 water molecules per each dean molecule at the interface, which do not take part in formation of hydrogen bonds, and a sharp radius decrease (from 100 to 20 nm of ice crystals formed in cell suspension at its freezing. The results could give rise to safety improving of their cells at their cryopreservation and low temperature storage conditions by incorporating into a powder composite environment.

  3. Hematologic and serum chemistry reference intervals for free-ranging lions (Panthera leo).

    Science.gov (United States)

    Maas, Miriam; Keet, Dewald F; Nielen, Mirjam

    2013-08-01

    Hematologic and serum chemistry values are used by veterinarians and wildlife researchers to assess health status and to identify abnormally high or low levels of a particular blood parameter in a target species. For free-ranging lions (Panthera leo) information about these values is scarce. In this study 7 hematologic and 11 serum biochemistry values were evaluated from 485 lions from the Kruger National Park, South Africa. Significant differences between sexes and sub-adult (≤ 36 months) and adult (>36 months) lions were found for most of the blood parameters and separate reference intervals were made for those values. The obtained reference intervals include the means of the various blood parameter values measured in captive lions, except for alkaline phosphatase in the subadult group. These reference intervals can be utilized for free-ranging lions, and may likely also be used as reference intervals for captive lions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Serum-Free Media and the Immunoregulatory Properties of Mesenchymal Stem Cells In Vivo and In Vitro

    OpenAIRE

    Mei Wu; Zhi-Bo Han; Jun Feng Liu; You Wei Wang; Jian Zhong Zhang; Chun Tuan Li; Peng Liang Xin; Zhong Chao Han; Xiong Peng Zhu

    2014-01-01

    Background: Mesenchymal stem cells are capable of self-renewal and multi-lineage differentiation. They are used extensively to treat several diseases. Traditionally, mesenchymal stem cells are cultured in serum-containing media, typically supplemented with fetal bovine serum (FBS). However, the variability of FBS is likely to skew experimental results. Although serum-free media used to expand mesenchymal stem cells has facilitated remarkable achievements, immunomodulation of these cells in un...

  5. Incorporation of [14C]-palmitate into lipids of Brassica cells during the induction of freezing tolerance

    International Nuclear Information System (INIS)

    Lynch, D.V.; Joseph, R.A.

    1989-01-01

    Changes in plasma membrane lipid composition have been causally related to increased freezing tolerance. Studies of lipid metabolism during ABA induction of freezing tolerance in Brassica napus suspension cultures were undertaken. Cells were labeled with [ 14 C]-palmitate four days after transfer to fresh medium (control) or medium containing ABA (which increases freezing tolerance). At times between one and 20 hrs after labeling, ABA-treated cells incorporated almost twice the amount of label as controls cells. Approximately 80% of the radioactivity was associated with neutral lipids in ABA-treated cells and controls. Incorporation of label into total cellular polar lipids was 4.9 x 10 5 dpm/mg protein for control cells and 1 x 10 6 dpm/mg protein for cells transferred to medium containing ABA. Analysis of lipids following alkaline hydrolysis indicated that incorporation of [ 14 C]-palmitate into glucosylceramide of ABA-treated cells was less than 60% of control values when expressed relative to that of the total polar lipids. Incorporation into ceramides was also depressed in ABA-treated cells

  6. Optimized human platelet lysate as novel basis for a serum-, xeno-, and additive-free corneal endothelial cell and tissue culture.

    Science.gov (United States)

    Thieme, Daniel; Reuland, Lynn; Lindl, Toni; Kruse, Friedrich; Fuchsluger, Thomas

    2018-02-01

    The expansion of donor-derived corneal endothelial cells (ECs) is a promising approach for regenerative therapies in corneal diseases. To achieve the best Good Manufacturing Practice standard the entire cultivation process should be devoid of nonhuman components. However, so far, there is no suitable xeno-free protocol for clinical applications. We therefore introduce a processed variant of a platelet lysate for the use in corneal cell and tissue culture based on a Good Manufacturing Practice-grade thrombocyte concentrate. This processed human platelet lysate (phPL), free of any animal components and of anticoagulants such as heparin with a physiological ionic composition, was used to cultivate corneal ECs in vitro and ex vivo in comparison to standard cultivation with fetal calf serum (FCS). Human donor corneas were cut in quarters while 2 quarters of each cornea were incubated with the respective medium supplement. Three fields of view per quarter were taken into account for the analysis. Evaluation of phPL as a medium supplement in cell culture of immortalized EC showed a superior viability compared with FCS control with reduced cell proliferation. Furthermore, the viability during the expansion of primary cells is significantly (3-fold ±0.5) increased with phPL compared with FCS standard medium. Quartering donor corneas was traumatic for the endothelium and therefore resulted in increased EC loss. Interestingly, however, cultivation of the quartered pieces for 2 weeks in 0.1-mg/ml pHPL in Biochrome I showed a 21 (±10) % EC loss compared with 67 (±12) % EC loss when cultivated in 2% FCS in Biochrome I. The cell culture protocol with pHPL as FCS replacement seems to be superior to the standard FCS protocols with respect to EC survival. It offers a xeno-free and physiological environment for corneal endothelial cells. This alternative cultivation protocol could facilitate the use of EC for human corneal cell therapy. Copyright © 2017 John Wiley & Sons, Ltd.

  7. Evaluation of BSK-H Complete Medium Supplemented with Rabbit Serum and Sodium Bicarbonate for the Growth of Borrelia anserina

    Directory of Open Access Journals (Sweden)

    Bilal Aslam, Iftikhar Hussain*, Muhammad Shahid Mahmood and Ahrar Khan1

    2013-04-01

    Full Text Available The present study was conducted to compare the effect of 3 formulations Barbour-Stoenner-Kelly (BSK medium on the growth of Borrelia (B. anserina, the causative agent of avian borreliosis. Three different formulations of BSK medium (BSK-H, BSK-II and BSK-H complete medium were prepared. For the isolation of B. anserina, Argas ticks were inoculated in all the three (BSK-H, BSK-II and BSK-H complete formulations of BSK medium. All the samples were also observed for the impact of BSK medium on the growth (Generation time, Growth per hour, Specific growth rate of B. anserina. Phase contrast microscopy was performed for the observation of viable B. anserina cells, and additional confirmation of all the isolates was done by performing indirect immunofluorescence assay and PCR. BSK-H complete medium supplemented with 6% rabbit serum and sodium bicarbonate was found best when compared to two other formulations of BSK medium with respect to the isolation, generation time and growth rate of Borrelia spirochetes.

  8. Selection of osteoprogenitors from the jaw periosteum by a specific animal-free culture medium.

    Directory of Open Access Journals (Sweden)

    Dorothea Alexander

    Full Text Available The goal of our research work is to establish mesenchymal osteoprogenitors derived from human jaw periosteum for tissue engineering applications in oral and maxillofacial surgery. For future autologous and/or allogeneic transplantations, some issues must be addressed. On the one hand, animal-free culture conditions have yet to be established. On the other hand, attempts should be undertaken to shorten the in vitro culturing process efficiently. The aim of the present study is to compare and analyze the phenotype of osteoprogenitors from the jaw periosteum under normal FCS-containing and animal-free culture conditions. Therefore, we analyzed the proliferation rates of MesenCult-XF medium (MC- in comparison to DMEM-cultured JPCs. Whereas jaw periosteal cells (JPCs show relatively slow proliferation rates and a fibroblastoid shape under DMEM culture conditions, MC-cultured JPCs diminished their cell size significantly and proliferated rapidly. By live-monitoring measurements of adhesion and proliferation, we made an interesting observation: whereas the proliferation of the MSCA-1(+ subpopulation and the unseparated cell fraction were favored by the animal-free culture medium, the proliferation of the MSCA-1(- subpopulation seemed to be repressed under these conditions. The alkaline phosphatase expression pattern showed similar results under both culture conditions. Comparison of the mineralization capacity revealed an earlier formation of calcium-phosphate precipitates under MC culture conditions; however, the mineralization capacity of the DMEM-cultured cells seemed to be higher. We conclude that the tested animal-free medium is suitable for the in vitro expansion and even for the specific selection of osteoprogenitor cells derived from the jaw periosteum.

  9. Interactions between Skeletal Muscle Myoblasts and their Extracellular Matrix Revealed by a Serum Free Culture System.

    Science.gov (United States)

    Chaturvedi, Vishal; Dye, Danielle E; Kinnear, Beverley F; van Kuppevelt, Toin H; Grounds, Miranda D; Coombe, Deirdre R

    2015-01-01

    Decellularisation of skeletal muscle provides a system to study the interactions of myoblasts with muscle extracellular matrix (ECM). This study describes the efficient decellularisation of quadriceps muscle with the retention of matrix components and the use of this matrix for myoblast proliferation and differentiation under serum free culture conditions. Three decellularisation approaches were examined; the most effective was phospholipase A2 treatment, which removed cellular material while maximizing the retention of ECM components. Decellularised muscle matrices were then solubilized and used as substrates for C2C12 mouse myoblast serum free cultures. The muscle matrix supported myoblast proliferation and differentiation equally as well as collagen and fibronectin. Immunofluorescence analyses revealed that myoblasts seeded on muscle matrix and fibronectin differentiated to form long, well-aligned myotubes, while myoblasts seeded on collagen were less organized. qPCR analyses showed a time dependent increase in genes involved in skeletal muscle differentiation and suggested that muscle-derived matrix may stimulate an increased rate of differentiation compared to collagen and fibronectin. Decellularized whole muscle three-dimensional scaffolds also supported cell adhesion and spreading, with myoblasts aligning along specific tracts of matrix proteins within the scaffolds. Thus, under serum free conditions, intact acellular muscle matrices provided cues to direct myoblast adhesion and migration. In addition, myoblasts were shown to rapidly secrete and organise their own matrix glycoproteins to create a localized ECM microenvironment. This serum free culture system has revealed that the correct muscle ECM facilitates more rapid cell organisation and differentiation than single matrix glycoprotein substrates.

  10. An improved method for the radioimmunoassay of free-thyroxine in serum dialysates

    International Nuclear Information System (INIS)

    Giles, A.F.

    1982-01-01

    A convenient, sensitive radioimmunoassay (using 125 I) of free thyroxine in serum dialysates is described. The method utilizes a solid phase separation system (pre-formed double antibody) and a relatively short incubation period (220 min) with a staggered addition of tracer. Blanks were low and consistent. The normal range for non-pregnant euthyroid samples (n = 59) was 11-23 pmol/l. Third trimester pregnancy samples were mostly within the normal range but at the lower end. Patients on T4 replacement showed a much wider variation in free T4 levels with many samples in the hyperthyroid region. Some hypothyroid samples had undetectable free T4 levels and hyperthyroid samples were frequently greater than 80 pmol/l. (author)

  11. The influence of metronidazole on free thymidine content of blood serum of irradiated mice

    International Nuclear Information System (INIS)

    Konov, A.V.; Ryabchenko, N.I.

    1986-01-01

    The influence of a radiosensitizer, metronidazole, on the free thymidine content of blood serum of irradiated mice was studied in aerobic and hypoxic conditions. A heated metronidazole solution (1 mg/g) was administered intraperitoneally 30 min before irradiation of animals with a dose of 3 Gy. Thymidine concentration in blood serum was determined by the radioimmunological technique. The influence of metronidazole on the level of thymidinemia was only noted in the animals exposed under hypoxic conditions

  12. Interaction of estradiol and high density lipoproteins on proliferation of the human breast cancer cell line MCF-7 adapted to grow in serum free conditions

    International Nuclear Information System (INIS)

    Jozan, S.; Faye, J.C.; Tournier, J.F.; Tauber, J.P.; David, J.F.; Bayard, F.

    1985-01-01

    The responsiveness of the human mammary carcinoma cell line MCF-7 to estradiol and tamoxifen treatment has been studied in different culture conditions. Cells from exponentially growing cultures were compared with cells in their initial cycles after replating from confluent cultures (''confluent-log'' cells). It has been observed that estradiol stimulation of tritiated thymidine incorporation decreases with cell density and that ''confluent-log'' cells are estrogen unresponsive for a period of four cell cycles in serum-free medium conditions. On the other hand, growth of cells replated from exponentially growing, as well as from confluent cultures, can be inhibited by tamoxifen or a combined treatment with tamoxifen and the progestin levonorgestrel. This growth inhibitory effect can be rescued by estradiol when cells are replated from exponentially growing cultures. The growth inhibitory effect cannot be rescued by estradiol alone (10(-10) to 10(-8) M) when cells are replated from confluent cultures. In this condition, the addition of steroid depleted serum is necessary to reverse the state of estradiol unresponsiveness. Serum can be replaced by high density lipoproteins but not by low density lipoproteins or lipoprotein deficient serum. The present data show that estradiol and HDL interact in the control of MCF-7 cell proliferation

  13. Comparative Serum Fatty Acid Profiles of Captive and Free-Ranging Cheetahs (Acinonyx jubatus) in Namibia.

    Science.gov (United States)

    Tordiffe, Adrian S W; Wachter, Bettina; Heinrich, Sonja K; Reyers, Fred; Mienie, Lodewyk J

    2016-01-01

    Cheetahs (Acinonyx jubatus) are highly specialised large felids, currently listed as vulnerable on the IUCN red data list. In captivity, they are known to suffer from a range of chronic non-infectious diseases. Although low heterozygosity and the stress of captivity have been suggested as possible causal factors, recent studies have started to focus on the contribution of potential dietary factors in the pathogenesis of these diseases. Fatty acids are an important component of the diet, not only providing a source of metabolisable energy, but serving other important functions in hormone production, cellular signalling as well as providing structural components in biological membranes. To develop a better understanding of lipid metabolism in cheetahs, we compared the total serum fatty acid profiles of 35 captive cheetahs to those of 43 free-ranging individuals in Namibia using gas chromatography-mass spectrometry. The unsaturated fatty acid concentrations differed most remarkably between the groups, with all of the polyunsaturated and monounsaturated fatty acids, except arachidonic acid and hypogeic acid, detected at significantly lower concentrations in the serum of the free-ranging animals. The influence of age and sex on the individual fatty acid concentrations was less notable. This study represents the first evaluation of the serum fatty acids of free-ranging cheetahs, providing critical information on the normal fatty acid profiles of free-living, healthy individuals of this species. The results raise several important questions about the potential impact of dietary fatty acid composition on the health of cheetahs in captivity.

  14. Serum lutein response is greater from free lutein than from esterified lutein during 4 weeks of supplementation in healthy adults.

    Science.gov (United States)

    Norkus, Edward P; Norkus, Katherine L; Dharmarajan, T S; Schierle, Joseph; Schalch, Wolfgang

    2010-12-01

    Current data suggest great variability in serum response following lutein ingestion from various sources. To compare the relative serum response during supplementation with free lutein (fL) and lutein esters (Le). 72 volunteers (23-52 years; body mass index [BMI] >20 and lutein lutein or 27 mg of lutein ester (equivalent to 13.5 mg free lutein), respectively. Fasting blood was obtained at baseline and after 7, 14, 21, and 28 days of supplementation. Supplements were consumed with standard portions of dry, ready-to-eat cereal and 2% cow's milk. Absolute changes in serum lutein, per mg daily dose, were significantly greater in fL vs. Le after 21 days (p  =  0.0012) and remained so after 28 days (p  =  0.0011) of supplementation. Serum lutein Area Under the Curve [AUC((day 0-28))] response was 17% greater for fL vs. Le (p  =  0.0187). Regression models were used and determined that (1) baseline serum lutein levels and (2) the form of lutein ingested (fL > Le) influence the serum lutein response during supplementation, while subject age, gender, BMI, and serum lipids do not affect serum response. These results suggest that the relative serum lutein response will be significantly greater from supplements containing free lutein than from supplements containing lutein esters. These findings should be useful for future clinical trials exploring the effectiveness of lutein supplementation in the prevention of or protection against age-related macular degeneration and/or cataracts.

  15. Immersion and contact freezing experiments in the Mainz wind tunnel laboratory

    Science.gov (United States)

    Eppers, Oliver; Mayer, Amelie; Diehl, Karoline; Mitra, Subir; Borrmann, Stephan; Szakáll, Miklós

    2016-04-01

    Immersion and contact freezing are of outmost important ice nucleation processes in mixed phase clouds. Experimental studies are carried out in the Mainz vertical wind tunnel laboratory in order to characterize these nucleation processes for different ice nucleating particles (INP), such as for mineral dust or biological particles. Immersion freezing is investigated in our laboratory with two different experimental techniques, both attaining contact-free levitation of liquid droplets and cooling of the surrounding air down to about -25 °C. In an acoustic levitator placed in the cold room of our laboratory, drops with diameters of 2 mm are investigated. In the vertical air stream of the wind tunnel droplets with diameter of 700 micron are freely floated at their terminal velocities, simulating the flow conditions of the free atmosphere. Furthermore, the wind tunnel offers a unique platform for contact freezing experiments. Supercooled water droplets are floated in the vertical air stream at their terminal velocities and INP are injected into the tunnel air stream upstream of them. As soon as INP collides with the supercooled droplet the contact freezing is initiated. The first results of immersion and contact freezing experiments with cellulose particles both in the acoustic levitator and in the wind tunnel will be presented. Cellulose is considered as typical INP of biological origin and a macrotracer for plant debris. Nucleating properties of cellulose will be provided, mainly focusing on the temperature, INP concentration, and specific surface area dependences of the freezing processes. Direct comparison between the different experimental techniques (acoustic levitator and wind tunnel), as well as between nucleation modes (immersion and contact freezing) will be presented. The work is carried out within the framework of the German research unit INUIT.

  16. [Diagnostic significance of serum free DNA human telomerase reverse transcriptase quantitative determination on spinal cord injury].

    Science.gov (United States)

    Yang, M K; Tang, J; Xiang, Z; Zhang, X; Wang, J; Li, Z; Li, Y; Sheng, W B

    2018-02-06

    Objective: To investigate the relationship between the content of human telomerase reverse transcriptase (hTERT) and its clinical features in serum free DNA in patients with different degree of spinal cord injury. Methods: From December 2013 to December 2016, inpatients of the Central Hospital of Bazhong, Sichuan Province were enrolledand divided into the experimental group, the disease control group and the negative control group. For the experimental group: 46 patients with spinal cord injury were graded according to the criteria of the American Association of Spinal Cord Injury (ASIA), including 12 cases of grade A, 10 cases of grade B, 10 cases of grade C, 7 cases of grade D and 7 cases of grade E; for the disease control group: 15 patients with spinal fractures (without spinal cord injury) at the same period were included; and for the negative control group: 20 healthy adult volunteers aged 18-50 years were selected.Real-time fluorescence quantitative PCR and immunoblotting were performed to detect the content of hTERT in serum free DNA both in patients and healthy controls and to compare the difference between them. The results of the somatosensory evoked potential (SEP) of all patients were compared and analyzed.The receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of hTERT content in serum free DNA in patients with spinal cord injury. Results: Comparison of serum free DNA hTERT content: in the experimental group, the serum free DNA hTERT content of grade A, B, C, D, E was (99.63±8.23), (76.24±4.37), (46.07±5.43), (16.30±0.95) and (15.74±1.12)μg/L, respectively.While it was (15.01±1.39)μg/L in the disease control group and (14.54±1.03)μg/L in the negative control group. The total difference was statistically significant between patients of each group and the control group ( F =857.917, P spinal cord injury has a certain guiding significance for the diagnosis of spinal cord injury and the degree of injury.

  17. Design and Characterization of DNA Strand-Displacement Circuits in Serum-Supplemented Cell Medium.

    Science.gov (United States)

    Fern, Joshua; Schulman, Rebecca

    2017-09-15

    The functional stability and lifetimes of synthetic molecular circuits in biological environments are important for long-term, stable sensors or controllers of cell or tissue behavior. DNA-based molecular circuits, in particular DNA strand-displacement circuits, provide simple and effective biocompatible control mechanisms and sensors, but are vulnerable to digestion by nucleases present in living tissues and serum-supplemented cell culture. The stability of double-stranded and single-stranded DNA circuit components in serum-supplemented cell medium and the corresponding effect of nuclease-mediated degradation on circuit performance were characterized to determine the major routes of degradation and DNA strand-displacement circuit failure. Simple circuit design choices, such as the use of 5' toeholds within the DNA complexes used as reactants in the strand-displacement reactions and the termination of single-stranded components with DNA hairpin domains at the 3' termini, significantly increase the functional lifetime of the circuit components in the presence of nucleases. Simulations of multireaction circuits, guided by the experimentally measured operation of single-reaction circuits, enable predictive realization of multilayer and competitive-reaction circuit behavior. Together, these results provide a basic route to increased DNA circuit stability in cell culture environments.

  18. Freezing for Love

    DEFF Research Database (Denmark)

    Carroll, Katherine; Kroløkke, Charlotte

    2018-01-01

    The promise of egg freezing for women’s fertility preservation entered feminist debate in connection with medical and commercial control over, and emancipation from, biological reproduction restrictions. In this paper we explore how women negotiate and make sense of the decision to freeze...... their eggs. Our analysis draws on semi-structured interviews with 16 women from the Midwest and East Coast regions of the USA who froze their eggs. Rather than freezing to balance career choices and ‘have it all’, the women in this cohort were largely ‘freezing for love’ and in the hope of having their ‘own...... healthy baby’. This finding extends existing feminist scholarship and challenges bioethical concerns about egg freezing by drawing on the voices of women who freeze their eggs. By viewing egg freezing as neither exclusively liberation nor oppression or financial exploitation, this study casts egg freezing...

  19. Active Stat3 is required for survival of human squamous cell carcinoma cells in serum-free conditions

    Directory of Open Access Journals (Sweden)

    DiGiovanni John

    2006-04-01

    Full Text Available Abstract Background Squamous cell carcinoma (SCC of the skin is the most aggressive form of non-melanoma skin cancer (NMSC, and is the single most commonly diagnosed cancer in the U.S., with over one million new cases reported each year. Recent studies have revealed an oncogenic role of activated signal transducer and activator of transcription 3 (Stat3 in many human tumors, especially in those of epithelial origin, including skin SCC. Stat3 is a mediator of numerous growth factor and cytokine signaling pathways, all of which activate it through phosphorylation of tyrosine 705. Results To further address the role of Stat3 in skin SCC tumorigenesis, we have analyzed a panel of human skin-derived cell lines ranging from normal human epidermal keratinocytes (NHEK, to non-tumorigenic transformed skin cells (HaCaT, to highly tumorigenic cells (SRB1-m7 and SRB12-p9 and observed a positive correlation between Stat3 phosphorylation and SCC malignancy. We next determined the role of Stat3 activity in cell proliferation and viability under serum-free culture conditions. This was accomplished by suppressing Stat3 activity in the SRB12-p9 cells through stable expression of a dominant negative acting form of Stat3β, which contains a tyrosine 705 to phenylalanine mutation (S3DN. The S3DN cells behaved similar to parental SRB12-p9 cells when cultured in optimal growth conditions, in the presence of 10% fetal calf serum. However, unlike the SRB12-p9 cells, S3DN cells underwent apoptotic cell death when cultured in serum-free medium (SFM. This was evidenced by multiple criteria, including accumulation of sub-G1 particles, induced PARP cleavage, and acquisition of the characteristic morphological changes associated with apoptosis. Conclusion This study provides direct evidence for a role for Stat3 in maintaining cell survival in the conditions of exogenous growth factor deprivation produced by culture in SFM. We also propose that delivery of the S3DN gene or

  20. Serum Levels Of Free And Total Insulin-Like Growth Factor (IGF)-1 And IGF Binding Protein-3 In Normal And Growth Hormone Deficient Children

    International Nuclear Information System (INIS)

    Shousha, M.A.; Soliman, S.E.T.; Hafez, M.H.

    2006-01-01

    Serum levels of total insulin-like growth factor-1 (IGF-1) and IGF-binding protein-3 (IGFBP-3) reflect the endogenous GH secretion in healthy children, which makes them good diagnostic markers for screening growth hormone deficiency (GHD) in short children, although some controversy still exists. Only a minor fraction of the total IGF-1 circulates in its free form, which is believed to be the biologically active form. Serum levels of free IGF-1, total IGF-I and IGFBP-3 were measured in 144 healthy children (72 boys and 72 girls, aged from 0 to 16 years) and in 12 pre-pubertal GH deficient (GHD) children to study the correlation between the age and free IGF-1, total IGF-1 and IGFBP-3 levels. In healthy subjects (both sexes), serum free IGF-1, total IGF-1 and IGFBP-3 levels were low in infancy, increasing during puberty and declining thereafter. Free IGF-1 in serum occupied about 0.97-1.45 % of the total IGF-1 values, and the ratios of free IGF-1 to total IGF-1 were significantly increased in the pubertal age groups than in the pre-pubertal age groups. Serum levels of free IGF-1 showed significant positive correlation with those of total IGF-I and IGFBP-3. Serum free IGF-1, total IGF-1 and IGFBP-3 levels in patients with GHD were decreased significantly with increasing the degree of hypopituitarism. These observations suggest that the increase in serum free IGF-1 level during puberty was caused by a dramatic increase in total IGF-1 rather than IGFBP-3. Also, high levels of these hormones may play an important role in pubertal growth spurt and may become a useful tool for diagnosing GHD and predicting growth response to long term GH therapy

  1. Serum levels of free and total insulin-link growth factor (IGF)-1 and (IGF) binding protein-3 in normal and growth hormone deficient children

    International Nuclear Information System (INIS)

    Shousha, M.A.; Soliman, S.E.T.; Hafez, H.M.

    2008-01-01

    Serum levels of total insulin-like growth factor- 1 (IGF-1) and IGF-binding protein-3 (IGFBP-3) reflect endogenous GH secretion in healthy children, which makes them good diagnostic markers for screening GH deficiency (GHD) in short children, although some controversy still exists. Only a minor fraction of the total IGF-1 circulates in its free form, which is believed to be the biologically active form. Serum levels of free IGF-1, total IGF-I and IGFBP-3 were measured in 144 healthy children (72 boys and 72 girls, aged from 0 to 16 years) and in 12 prepubertal GH. deficient (GHD) children to study correlation between the age and free IGF-1, total IGF-1 and IGFBP-3 levels. In healthy subjects (both sexes), serum free IGF-1, total IGF-1 and IGFBP-3 levels were low in infancy, increasing during puberty and declining thereafter. Free IGF-1 in serum occupied about 0.97. 1.45 % of the total IGF-1 values, and the ratios of free IGF-1 to total IGF-1 were significantly increased in the pubertal age groups than in the prepubertal age groups. Serum levels of free IGF-1 showed significant positive correlation with those of total IGF-I and IGFBP-3. Serum free IGF-1, total IGF-1 and IGFBP-3 levels in patients with GHD decreased significantly with increasing degree of hypopituitarism. These observations suggest that the increase in serum free IGF-1 level during puberty was caused by a dramatic increase in total IGF-1 rather than IGFBP-3. Also, high levels of these hormones may play an important role in pubertal growth spurt and may become a useful tool for diagnosing GHD and predicting growth response to long term GH therapy

  2. Influence of freezing down to 77.15 K on structure and antioxidant power of some proteins

    Science.gov (United States)

    Rozanova, S. L.; Narozhnyi, S. V.; Nardid, O. A.

    2017-06-01

    The purpose of the present work was to investigate influence of different freeze-thawing protocols on structure and antioxidant properties of isolated proteins. In our experiments we have studied human serum albumin, human hemoglobin and cytochrome C derived from equine heart frozen down to 77.15 K with 1-2 deg/min and 300 deg/min rate with following thawing on a water bath at 293.15 K. Native proteins were assumed as a control. Influence of freeze-thawing protocols on protein structure was investigated using spectrophotometric and fluorescent assays. Antioxidant activities of isolated proteins were estimated by their ability to reduce ABTS+ radical. It has been established that unfolding derived from freeze-thawing exposure leads to protein antioxidant activity increasing while decreasing of such an activity may be connected with macromolecule aggregation. Character of freeze-thawing influence on antioxidant activity of proteins depends on molecule structure peculiarities and freezing protocols.

  3. Freezing Bubbles

    Science.gov (United States)

    Kingett, Christian; Ahmadi, Farzad; Nath, Saurabh; Boreyko, Jonathan

    2017-11-01

    The two-stage freezing process of a liquid droplet on a substrate is well known; however, how bubbles freeze has not yet been studied. We first deposited bubbles on a silicon substrate that was chilled at temperatures ranging from -10 °C to -40 °C, while the air was at room temperature. We observed that the freeze front moved very slowly up the bubble, and in some cases, even came to a complete halt at a critical height. This slow freezing front propagation can be explained by the low thermal conductivity of the thin soap film, and can be observed more clearly when the bubble size or the surface temperature is increased. This delayed freezing allows the frozen portion of the bubble to cool the air within the bubble while the top part is still liquid, which induces a vapor pressure mismatch that either collapses the top or causes the top to pop. In cases where the freeze front reaches the top of the bubble, a portion of the top may melt and slowly refreeze; this can happen more than just once for a single bubble. We also investigated freezing bubbles inside of a freezer where the air was held at -20 °C. In this case, the bubbles freeze quickly and the ice grows radially from nucleation sites instead of perpendicular to the surface, which provides a clear contrast with the conduction limited room temperature bubbles.

  4. Prognostic Value of Serum Free Light Chain in Multiple Myeloma.

    Science.gov (United States)

    El Naggar, Amel A; El-Naggar, Mostafa; Mokhamer, El-Hassan; Avad, Mona W

    2015-01-01

    The measurement of serum free light chain (sFLC) has been shown to be valuable in screening for the presence of plasma cell dyscrasia as well as for baseline prognosis in newly diagnosed patients. The aim of the present work was to study the prognostic value of sFLC in multiple myeloma in relation to other serum biomarkers, response to therapy and survival. Forty five newly diagnosed patients with MM were included in the study. Patients were divided into responders and non-responders groups according to response to therapy. sFLC and serum Amyloid A (SAA) were measured by immunonephelometry. The non-responders group showed a statistically significant higher kappa/lambda or lambda/kappa ratio and higher β2 microglobulin level, but lower albumin level at presentation, as compared to the responders group (P < 0.001). However, no statistically significant difference was detected between the two groups regarding SA A or calcium levels. Comparison between sFLC ratio obtained before and after therapy revealed significant decrease after treatment in the responders group (P = 0.05). Survival was significantly inferior in patients with an FLC ratio of ≥ 2.6 or ≤ 0.56 compared with those with an FLC ratio that was between 0.56 and 2.6 (P = 0.002).

  5. Comparative Serum Fatty Acid Profiles of Captive and Free-Ranging Cheetahs (Acinonyx jubatus) in Namibia

    Science.gov (United States)

    Wachter, Bettina; Heinrich, Sonja K.; Reyers, Fred; Mienie, Lodewyk J.

    2016-01-01

    Cheetahs (Acinonyx jubatus) are highly specialised large felids, currently listed as vulnerable on the IUCN red data list. In captivity, they are known to suffer from a range of chronic non-infectious diseases. Although low heterozygosity and the stress of captivity have been suggested as possible causal factors, recent studies have started to focus on the contribution of potential dietary factors in the pathogenesis of these diseases. Fatty acids are an important component of the diet, not only providing a source of metabolisable energy, but serving other important functions in hormone production, cellular signalling as well as providing structural components in biological membranes. To develop a better understanding of lipid metabolism in cheetahs, we compared the total serum fatty acid profiles of 35 captive cheetahs to those of 43 free-ranging individuals in Namibia using gas chromatography-mass spectrometry. The unsaturated fatty acid concentrations differed most remarkably between the groups, with all of the polyunsaturated and monounsaturated fatty acids, except arachidonic acid and hypogeic acid, detected at significantly lower concentrations in the serum of the free-ranging animals. The influence of age and sex on the individual fatty acid concentrations was less notable. This study represents the first evaluation of the serum fatty acids of free-ranging cheetahs, providing critical information on the normal fatty acid profiles of free-living, healthy individuals of this species. The results raise several important questions about the potential impact of dietary fatty acid composition on the health of cheetahs in captivity. PMID:27992457

  6. Cell growth and resistance of Lactococcus lactis subsp. lactis TOMSC161 following freezing, drying and freeze-dried storage are differentially affected by fermentation conditions.

    Science.gov (United States)

    Velly, H; Fonseca, F; Passot, S; Delacroix-Buchet, A; Bouix, M

    2014-09-01

    To investigate the effects of fermentation parameters on the cell growth and on the resistance to each step of the freeze-drying process of Lactococcus lactis subsp. lactis TOMSC161, a natural cheese isolate, using a response surface methodology. Cells were cultivated at different temperatures (22, 30 and 38°C) and pH (5·6, 6·2 and 6·8) and were harvested at different growth phases (0, 3 and 6 h of stationary phase). Cultivability and acidification activity losses of Lc. lactis were quantified after freezing, drying, 1 and 3 months of storage at 4 and 25°C. Lactococcus lactis was not damaged by freezing but was sensitive to drying and to ambient temperature storage. Moreover, the fermentation temperature and the harvesting time influenced the drying resistance of Lc. lactis. Lactococcus lactis cells grown in a whey-based medium at 32°C, pH 6·2 and harvested at late stationary phase exhibited both an optimal growth and the highest resistance to freeze-drying and storage. A better insight on the individual and interaction effects of fermentation parameters made it possible the freeze-drying and storage preservation of a sensitive strain of technological interest. Evidence on the particularly damaging effect of the drying step and the high-temperature storage is presented. © 2014 The Society for Applied Microbiology.

  7. ADAM-12 stability in first trimester maternal serum.

    Science.gov (United States)

    Cowans, N J; Stamatopoulou, A; Jaakohuhta, S; Spencer, K

    2010-06-01

    Maternal serum A Disintergrin And Metaloprotease-12 (ADAM-12) has been proposed as a marker for prenatal screening of chromosomal abnormalities, pre-eclampsia and other adverse pregnancy outcomes. In this study, we examine the stability of ADAM-12 with time and at different temperatures. Maternal serum and whole blood pools were stored at 30 degrees C, room temperature and refrigerator temperature or subjected to repeated freeze-thaw cycles. ADAM-12 was measured at set time points using an automated DELFIA research assay. Using a 10% change in concentration as a limit of stability, ADAM-12 is stable in serum for less than 15 h at 30 degrees C, less than 20 h at room temperature and for 51 h at refrigerator temperature. ADAM-12 levels are not altered following three - 20 degrees C to room temperature freeze-thaw cycles. The stability of ADAM-12 in whole blood appears similar to that in serum. The findings of this study suggest that ADAM-12 may be unstable under many routine laboratory conditions, and the marker's instability may also be partly responsible for the discrepancies in the literature.

  8. Method validation for preparing serum and plasma samples from human blood for downstream proteomic, metabolomic, and circulating nucleic acid-based applications.

    Science.gov (United States)

    Ammerlaan, Wim; Trezzi, Jean-Pierre; Lescuyer, Pierre; Mathay, Conny; Hiller, Karsten; Betsou, Fay

    2014-08-01

    Formal method validation for biospecimen processing in the context of accreditation in laboratories and biobanks is lacking. Serum and plasma processing protocols were validated for fitness-for-purpose in terms of key downstream endpoints, and this article demonstrates methodology for biospecimen processing method validation. Serum and plasma preparation from human blood was optimized for centrifugation conditions with respect to microparticle counts. Optimal protocols were validated for methodology and reproducibility in terms of acceptance criteria based on microparticle counts, DNA and hemoglobin concentration, and metabolomic and proteomic profiles. These parameters were also used to evaluate robustness for centrifugation temperature (4°C versus room temperature [RT]), deceleration (low, medium, high) and blood stability (after a 2-hour delay). Optimal protocols were 10-min centrifugation for serum and 20-min for plasma at 2000 g, medium brake, RT. Methodology and reproducibility acceptance criteria were met for both protocols except for reproducibility of plasma metabolomics. Overall, neither protocol was robust for centrifugation at 4°C versus RT. RT gave higher microparticles and free DNA yields in serum, and fewer microparticles with less hemolysis in plasma. Overall, both protocols were robust for fast, medium, and low deceleration, with a medium brake considered optimal. Pre-centrifugation stability after a 2-hour delay was seen at both temperatures for hemoglobin concentration and proteomics, but not for microparticle counts. We validated serum and plasma collection methods suitable for downstream protein, metabolite, or free nucleic acid-based applications. Temperature and pre-centrifugation delay can influence analytic results, and laboratories and biobanks should systematically record these conditions in the scope of accreditation.

  9. Proposal of a candidate international conventional reference measurement procedure for free thyroxine in serum.

    NARCIS (Netherlands)

    Thienpont, L.M.; Beastall, G.H.; Christofides, N.D.; Faix, J.D.; Ieiri, T.; Jarrige, V.; Miller, W.G.; Miller, R.; Nelson, J.C.; Ronin, C.; Ross, H.A.; Rottmann, M.; Thijssen, J.H.; Toussaint, B.

    2007-01-01

    In the present paper the IFCC WG-STFT recommends and provides the rationale to establish metrological traceability of serum free thyroxine (FT4) measurements to a candidate international conventional reference measurement procedure. It is proposed that this procedure be based on equilibrium dialysis

  10. Freezing in the presence of disorder: a lattice study

    International Nuclear Information System (INIS)

    Schmidt, Matthias; Lafuente, Luis; Cuesta, Jose A

    2003-01-01

    We investigate the freezing transition in a two-dimensional lattice model of annealed hard squares that are subject to the influence of randomly placed quenched particles of the same size. The latter model is a porous medium. By combining two recent density functional approaches we arrive at a theory for quenched-annealed lattice fluids that treats the quenched particles on the level of their one-body density distribution. We show that this approach yields thermodynamics that compare well with results from treating matrix realizations explicitly and performing subsequent averaging over the disorder. The freezing transition from a fluid to a columnar phase is found to be continuous. On increasing matrix density it shifts towards close packing and vanishes beyond a threshold matrix density

  11. Viability of G4 after Spray-Drying and Freeze-Drying

    Directory of Open Access Journals (Sweden)

    Stephenie Wong

    2010-01-01

    Full Text Available Viability of Bifidobacterium pseudocatenulatum G4 following spray-drying and freeze-drying in skim milk was evaluated. After spray-drying, the strain experienced over 99% loss in viability regardless of the air outlet temperature (75 and 85 °C and the heat-adaptation temperature (45 and 65 °C, 30 min. The use of heat-adaptation treatment to improve the thermotolerance of this strain was ineffective. On the other hand, the strain showed a superior survival at 71.65%–82.07% after freeze-drying. Viable populations of 9.319–9.487 log 10 cfu/g were obtained when different combinations of skim milk and sugar were used as cryoprotectant. However, the addition of sugars did not result in increased survival during the freeze-drying process. Hence, 10% (w/v skim milk alone is recommended as a suitable protectant and drying medium for this strain. The residual moisture content obtained was 4.41% ± 0.44%.

  12. Essential amino acids in the gluten-free diet and serum in relation to depression in patients with celiac disease.

    Directory of Open Access Journals (Sweden)

    Nathalie J M van Hees

    Full Text Available Celiac disease (CD is associated with an increased risk of major depressive disorder, possibly due to deficiencies in micronutrients in the gluten-free diet. We aimed to investigate whether essential amino acids (i.e., the precursors of serotonin, dopamine and other neurotransmitters are depleted in the diet and serum of CD patients with major depressive disorder.In a cross-sectional study we assessed dietary intake of amino acids and serum levels of amino acids, in 77 CD patients on a gluten-free diet and in 33 healthy controls. Major depressive disorder was assessed with structured interviews (using the Mini International Neuropsychiatric Interview Plus. Dietary intake was assessed using a 203-item food frequency questionnaire.Participants had a mean age of 55 years and 74% were women. The intake of vegetable protein was significantly lower in CD patients than in healthy controls (mean difference of 7.8 g/d; 95% CI: 4.7-10.8, as were serum concentrations of tyrosine, phenylalanine and tryptophan (all p < 0.005. However, within the CD patient group, the presence of major depressive disorder (n = 42 was not associated with intake or serum levels of essential amino acids.Patients with CD on a long-term gluten-free diet, with good adherence, consume significantly less vegetable protein than controls, and their serum levels of several essential amino acids were also lower. Despite its potential adverse effect, intake and serum levels of essential amino acids were not related to major depression.

  13. Analysis of Serum Total and Free PSA Using Immunoaffinity Depletion Coupled to SRM: Correlation with Clinical Immunoassay Tests

    Science.gov (United States)

    Liu, Tao; Hossain, Mahmud; Schepmoes, Athena A.; Fillmore, Thomas L.; Sokoll, Lori J.; Kronewitter, Scott R.; Izmirlian, Grant; Shi, Tujin; Qian, Wei-Jun; Leach, Robin J.; Thompson, Ian M.; Chan, Daniel W.; Smith, Richard D.; Kagan, Jacob; Srivastava, Sudhir; Rodland, Karin D.; Camp, David G.

    2012-01-01

    Recently, selected reaction monitoring mass spectrometry (SRM-MS) has been more frequently applied to measure low abundance biomarker candidates in tissues and biofluids, owing to its high sensitivity and specificity, simplicity of assay configuration, and exceptional multiplexing capability. In this study, we report for the first time the development of immunoaffinity depletion-based workflows and SRM-MS assays that enable sensitive and accurate quantification of total and free prostate-specific antigen (PSA) in serum without the requirement for specific PSA antibodies. Low ng/mL level detection of both total and free PSA was consistently achieved in both PSA-spiked female serum samples and actual patient serum samples. Moreover, comparison of the results obtained when SRM PSA assays and conventional immunoassays were applied to the same samples showed good correlation in several independent clinical serum sample sets. These results demonstrate that the workflows and SRM assays developed here provide an attractive alternative for reliably measuring candidate biomarkers in human blood, without the need to develop affinity reagents. Furthermore, the simultaneous measurement of multiple biomarkers, including the free and bound forms of PSA, can be performed in a single multiplexed analysis using high-resolution liquid chromatographic separation coupled with SRM-MS. PMID:22846433

  14. Validity of Serum Testosterone, Free Androgen Index, and Calculated Free Testosterone in Women with Suspected Hyperandrogenism

    Directory of Open Access Journals (Sweden)

    Manal K. Al Kindi

    2012-11-01

    Full Text Available Objectives: There are technical limitations for the currently available methods of measuring serum total and free testosteronein females. The study objectives were to evaluate the usefulness of serum total testosterone, sex hormone-binding globulin (SHBG, free androgen index (FAI, and calculated free testosterone (CFT in the assessment of androgen status in women investigated for suspected hyperandrogenism.Methods: This is a case control study that was conducted during the period from 1st May 2011 to 31st October 2011 on 122 patients aged (18-45 years whom were referred to the Clinical Biochemistry Laboratory from the Endocrinology and Gynecology Clinics, Royal Hospital, Oman. Women with no clinical feature or laboratory data indicative of hormonal dysfunction and with midluteal progesterone >30 nmol/L were selected as controls (group 1; n=18. The patients were divided into subgroups based on the clinical/laboratory diagnosis of polycystic ovary syndrome (PCOS [group 2; n=19, hirsutism (group 3; n=18, menstrual disturbances (irregularities or infertility (group 4; n=49, as well as combination of PCOS or hirsutism and menstrual disturbances or infertility (group 5;n=18. Serum total testosterone and SHBG were measured, FAI was calculated as percentage ratio of total testosterone to SHBG values, and CFT was calculated according to Vermeulen equation.Results: There was a statistically significant difference in the mean levels of testosterone, FAI and CFT in each patient group compared with the control group. For diagnosing hyperandrogenism, each indicator was selected at the recommended cut-off: testosterone >3.0 nmol/L, SHBG 5%, and CFT >32 pmol/L. In group 2, 89.5% and 94.7% of the patients had increased FAI and CFT, respectively; compared with 36.4% for increased testosterone. In group 3, 88.9% and 88.9% of the patients had similarly increased FAI and CFT, respectively; compared with 66.7% for testosterone. In group 4, patients had 63.3% and 73

  15. Human serum and platelet lysate are appropriate xeno-free alternatives for clinical-grade production of human MuStem cell batches.

    Science.gov (United States)

    Saury, Charlotte; Lardenois, Aurélie; Schleder, Cindy; Leroux, Isabelle; Lieubeau, Blandine; David, Laurent; Charrier, Marine; Guével, Laëtitia; Viau, Sabrina; Delorme, Bruno; Rouger, Karl

    2018-05-02

    Canine MuStem cells have demonstrated regenerative efficacy in a dog model of muscular dystrophy, and the recent characterization of human counterparts (hMuStem) has highlighted the therapeutic potential of this muscle-derived stem cell population. To date, these cells have only been generated in research-grade conditions. However, evaluation of the clinical efficacy of any such therapy will require the production of hMuStem cells in compliance with good manufacturing practices (GMPs). Because the current use of fetal bovine serum (FBS) to isolate and expand hMuStem cells raises several ethical, safety, and supply concerns, we assessed the use of two alternative xeno-free blood derivatives: human serum (HS) and a human platelet lysate (hPL). hMuStem cells were isolated and expanded in vitro in either HS-supplemented or hPL-supplemented media and the proliferation rate, clonogenicity, myogenic commitment potential, and oligopotency compared with that observed in FBS-supplemented medium. Flow cytometry and high-throughput 3'-digital gene expression RNA sequencing were used to characterize the phenotype and global gene expression pattern of hMuStem cells cultured with HS or hPL. HS-supplemented and hPL-supplemented media both supported the isolation and long-term proliferation of hMuStem cells. Compared with FBS-based medium, both supplements enhanced clonogenicity and allowed for a reduction in growth factor supplementation. Neither supplement altered the cell lineage pattern of hMuStem cells. In vitro differentiation assays revealed a decrease in myogenic commitment and in the fusion ability of hMuStem cells when cultured with hPL. In return, this reduction of myogenic potential in hPL-supplemented cultures was rapidly reversed by substitution of hPL with HS or fibrinogen-depleted hPL. Moreover, culture of hMuStem cells in hPL hydrogel and fibrinogen-depleted hPL demonstrated that myogenic differentiation potential is maintained in heparin-free hPL derivatives. Our

  16. Liposome encapsulated soy lecithin and cholesterol can efficiently replace chicken egg yolk in human semen cryopreservation medium.

    Science.gov (United States)

    Mutalik, Srinivas; Salian, Sujith Raj; Avadhani, Kiran; Menon, Jyothsna; Joshi, Haritima; Hegde, Aswathi Raju; Kumar, Pratap; Kalthur, Guruprasad; Adiga, Satish Kumar

    2014-06-01

    Cryopreservation of spermatozoa plays a significant role in reproductive medicine and fertility preservation. Chicken egg yolk is used as an extender in cryopreservation of human spermatozoa using glycerol egg yolk citrate (GEYC) buffered medium. Even though 50% survival of spermatozoa is generally achieved with this method, the risk of high levels of endotoxins and transmission pathogens from chicken egg yolk is a matter of concern. In the present study we attempted to establish a chemically defined cryopreservation medium which can replace the chicken egg yolk without affecting sperm survival. Ejaculates from 28 men were cryopreserved with GEYC based freezing medium or liposome encapsulated soy lecithin-cholesterol based freezing medium (LFM). The semen samples were subjected to rapid thawing after 14 days of storage in liquid nitrogen. Post-thaw analysis indicated significantly higher post-thaw motility and sperm survival in spermatozoa cryopreserved with LFM compared to conventional GEYC freezing medium. The soy lecithin and cholesterol at the ratio of 80:20 with sucrose showed the highest percentage of post-thaw motility and survival compared to the other compositions. In conclusion, chemically defined cryopreservation medium with liposome encapsulated soy lecithin and cholesterol can effectively replace the chicken egg yolk from human semen cryopreservation medium without compromising post-thaw outcome.

  17. Spray-freeze-drying of nanosuspensions: the manufacture of insulin particles for needle-free ballistic powder delivery.

    Science.gov (United States)

    Schiffter, Heiko; Condliffe, Jamie; Vonhoff, Sebastian

    2010-08-06

    The feasibility of preparing microparticles with high insulin loading suitable for needle-free ballistic drug delivery by spray-freeze-drying (SFD) was examined in this study. The aim was to manufacture dense, robust particles with a diameter of around 50 microm, a narrow size distribution and a high content of insulin. Atomization using ultrasound atomizers showed improved handling of small liquid quantities as well as narrower droplet size distributions over conventional two-fluid nozzle atomization. Insulin nanoparticles were produced by SFD from solutions with a low solid content (300 mg ml(-1)) consisting of trehalose, mannitol, dextran (10 kDa) and dextran (150 kDa) (abbreviated to TMDD) in order to maximize particle robustness and density after SFD. With the increase in insulin content, the viscosity of the nanosuspensions increased. Liquid atomization was possible up to a maximum of 250 mg of nano-insulin suspended in a 1.0 g matrix. However, if a narrow size distribution with a good correlation between theoretical and measurable insulin content was desired, no more than 150 mg nano-insulin could be suspended per gram of matrix formulation. Particles were examined by laser light diffraction, scanning electron microscopy and tap density testing. Insulin stability was assessed using size exclusion chromatography (SEC), reverse phase chromatography and Fourier transform infrared (FTIR) spectroscopy. Densification of the particles could be achieved during primary drying if the product temperature (T(prod)) exceeded the glass transition temperature of the freeze concentrate (T(g)') of -29.4 degrees C for TMDD (3331) formulations. Particles showed a collapsed and wrinkled morphology owing to viscous flow of the freeze concentrate. With increasing insulin loading, the d (v, 0.5) of the SFD powders increased and particle size distributions got wider. Insulin showed a good stability during the particle formation process with a maximum decrease in insulin monomer of

  18. Human Platelet Lysate as a Xeno Free Alternative of Fetal Bovine Serum for the In Vitro Expansion of Human Mesenchymal Stromal Cells.

    Science.gov (United States)

    Mohammadi, Saeed; Nikbakht, Mohsen; Malek Mohammadi, Ashraf; Zahed Panah, Mahdi; Ostadali, Mohammad Reza; Nasiri, Hajar; Ghavamzadeh, Ardeshir

    2016-07-01

    Mesenchymal stromal cells (MSCs) are employed in various different clinical settings in order to modulate immune response. Human autologous and allogeneic supplements including platelet derivatives such as platelet lysate (PL), platelet-released factors (PRF) and serum are assessed in clinical studies to replace fetal bovine serum (FBS). The immunosuppressive activity and multi-potential characteristic of MSCs appear to be maintained when the cells are expanded in platelet derivatives. Platelet-rich plasma was collected from umbrical cord blood (UCB). Platelet-derived growth factors obtained by freeze and thaw methods. CD62P expression was determined by flow cytometry. The concentration of PDGF-BB and PDGF-AB was detemined by ELISA. We tested the ability of a different concentration of PL-supplemented medium to support the ex vivo expansion of Wharton's jelly derived MSCs. We also investigated the biological/functional properties of expanded MSCs in presence of different concentration of PL. The conventional karyotyping was performed in order to study the chromosomal stability. The gene expression of Collagen I and II aggrecan and SOX-9 in the presence of different concentrations of PL was evaluated by Real-time PCR. We observed 5% and 10% PL, causing greater effects on proliferation of MSCs .These cells exhibited typical morphology, immunophenotype and differentiation capacity. The genetic stability of these derivative cells from Wharton's jelly was demonstrated by a normal karyotype. Furthermore, the results of Real-time PCR analysis showed that the expression of chondrocyte specific genes was higher in MSCs in the presence of 5% and 10% PL, compared with FBS supplement. We demonstrated that PL could be used as an alternative safe source of growth factors for expansion of MSCs and also maintained similar growing potential and phenotype without any effect on chromosomal stability.

  19. The principles of ultrasound and its application in freezing related processes of food materials: A review.

    Science.gov (United States)

    Cheng, Xinfeng; Zhang, Min; Xu, Baoguo; Adhikari, Benu; Sun, Jincai

    2015-11-01

    Ultrasonic processing is a novel and promising technology in food industry. The propagation of ultrasound in a medium generates various physical and chemical effects and these effects have been harnessed to improve the efficiency of various food processing operations. Ultrasound has also been used in food quality control as diagnostic technology. This article provides an overview of recent developments related to the application of ultrasound in low temperature and closely related processes such as freezing, thawing, freeze concentration and freeze drying. The applications of high intensity ultrasound to improve the efficiency of freezing process, to control the size and size distribution of ice crystals and to improve the quality of frozen foods have been discussed in considerable detail. The use of low intensity ultrasound in monitoring the ice content and to monitor the progress of freezing process has also been highlighted. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Freeze-substitution methods for Ni localization and quantitative analysis in Berkheya coddii leaves by means of PIXE

    International Nuclear Information System (INIS)

    Budka, D.; Mesjasz-PrzybyIowicz, J.; Tylko, G.; PrzybyIowicz, W.J.

    2005-01-01

    Leaves of Ni hyperaccumulator Berkheya coddii were chosen as a model to investigate the influence of eight freeze-substitution protocols on the Ni content and distribution. Freeze-substitution of leaf samples cryofixed by high-pressure freezing was carried out in dry acetone, methanol, diethyl ether and tetrahydrofuran. The same substitution media were also used with dimethylglyoxime added as a precipitation reagent. The samples were infiltrated and embedded in Spurr's resin. Micro-PIXE analysis of Ni concentration and localization, complemented by proton backscattering for matrix assessment, was performed using the nuclear microprobe at Materials Research Group, iThemba LABS, South Africa. True elemental maps and concentrations were obtained using GeoPIXE-II software. The results were compared with the control results obtained for the parallel air-dried samples, corrected for the water content. The highest Ni content was found in the leaf samples substituted in diethyl ether. This concentration was statistically different from the results obtained for other media. In case of diethyl ether medium Ni was mainly localized in the mesophyll tissue, and the distribution map of this element was in accordance with previous results obtained for freeze-dried and frozen-hydrated leaves of this species. The same distribution pattern was observed for specimens embedded in dry acetone, but Ni concentration was significantly lower. Tetrahydrofuran medium preserved Ni preferentially in the epidermis and vascular tissue, and the elemental map for samples embedded in this medium was distorted. Ni was almost completely washed out from samples substituted in methanol and it was thus impossible to obtain a picture of its distribution. Dimethylglyoxime did not improve the preservation of this element. These results show that diethyl ether is a suitable substitution medium for assessment of Ni concentration and distribution in leaves of B. coddii

  1. Clinical significance of two-step magnetic radioimmunoassay for determining serum free T3 and free T4

    International Nuclear Information System (INIS)

    Chen Bing

    1996-01-01

    The concentrations of the serum free T 3 (FT 3 ), free T 4 (FT 4 ), total triodothyronine (TT 3 ), total thyroxine (TT 4 ) and thyrotropin (TSH) are determined for 355 cases of normal persons, pregnant women and various thyropathetic patients. The normal values of FT 3 and FT 4 are 2.0-8.5 pmol/l, and 9.5-26.5 pmol/l, respectively. Neither FT 3 nor FT 4 is affected by the thyroxine combined with globulin (TBG), which is of unique diagnostic value for those with variable TBG (such as pregnant women hyperthyroidism, hypothyroidism, etc.), FT 3 and FT 4 are the most sensitive indices for diagnosis of hyperthyroidism, and hypothyroidism, respectively. In addition, FT 3 and FT 4 can greatly contribute to the observation of curative effectiveness under treatment

  2. The fluctuation of free amino acids in serum during acute ischemic stroke

    Directory of Open Access Journals (Sweden)

    Szpetnar Maria

    2016-12-01

    Full Text Available Currently, little data exists regarding the involvement of free amino acids (AA in the pathogenesis of ischemic stroke (IS. Thus, our objective was to study the degree of the degree of fluctuation of free amino acids level in serum during the acute phase of IS. The study consisted of eighteen patients (female/male: 10/8; age: 73.1 ± 4.1 with acute IS that was confirmed by way of computed tomography, while twelve sex and age matched individuals were assigned as control group. During the study period, the patients did not receive any supplemental amino acids therapy that could affect the obtained results. The venous blood was obtained after >3 hours fasting at two time-points; time-point 1 – at admission to the hospital; time-point 2 – on day 5 from stroke onset. The blood for control purposes was collected only once, and the blood collection at time-point 1 was done before thrombolytic treatment (nine patients. The amino acids were identified using the Amino Acids Analyser (AAA 400 by INGOS Corp., Praha, Czech Republic. Our results revealed a statistically significant increase of glutamate, cystine and methionine on day 1 of stroke, in comparison to control, whereas, proline level was decreased on day 1 of stroke – in comparison to control serum. On comparing day 5 to the initial day of IS, elevation was observed of levels of asparagine, glycine, tyrosine, arginine, threonine, valine, leucine and phenylalanine. It can be said, then, that ischemic stroke induces both essential and nonessential amino acid fluctuations. Moreover, the decrease in proline and glutamine serum level with the simultaneous increase in the concentration of branch chain amino acids, Glu and Thr suggests a violent mobilization of the body’s proteins. Thus, a decrease of Pro and a simultaneous increase of Glu serum level could be considered as a marker of acute IS.

  3. Combination treatment of clostridium perfringens spores to freezing and/or gamma irradiation

    International Nuclear Information System (INIS)

    El-Fouly, M.Z.; El-Zawahry, Y.A.; Aziz, N.H.

    1985-01-01

    Freezing process alone caused relatively low decrease in viable count of suspended spores in minced meat while it decreased the spore numbers suspended in saline solution by more than one log cycle especially in case of the Egyptian strain. An abrupt decrease in viable counts of clostridium spores was observed by application dose of 1KGY either before or after freezing followed by gradual decrease of viable counts up to 15 KGY. The synergestic effect of combined treatment was clearly obvious for spores suspended in minced meat, which usually contains protective agents which increase the resistance of microorganisms against the separate treatment of radiation of freezing especially with spores of NCTC 8798 strain. Freezing the saline suspending medium before or after irradiation after the sensitivity of clostridium spores by only small extent and gave negative synergestic effect in some treatment. The percentages of injured spores due to the combined treatment were ranged between 15-100% of the viable counts. The percentage of injured spores tended to increase as the radiation dose levels increased

  4. Saccharomyces cerevisiae glycerol/H+ symporter Stl1p is essential for cold/near-freeze and freeze stress adaptation. A simple recipe with high biotechnological potential is given

    Directory of Open Access Journals (Sweden)

    Ferreira Célia

    2010-11-01

    Full Text Available Abstract Background Freezing is an increasingly important means of preservation and storage of microbial strains used for many types of industrial applications including food processing. However, the yeast mechanisms of tolerance and sensitivity to freeze or near-freeze stress are still poorly understood. More knowledge on this regard would improve their biotechnological potential. Glycerol, in particular intracellular glycerol, has been assigned as a cryoprotectant, also important for cold/near-freeze stress adaptation. The S. cerevisiae glycerol active transporter Stl1p plays an important role on the fast accumulation of glycerol. This gene is expressed under gluconeogenic conditions, under osmotic shock and stress, as well as under high temperatures. Results We found that cells grown on STL1 induction medium (YPGE and subjected to cold/near-freeze stress, displayed an extremely high expression of this gene, also visible at glycerol/H+ symporter activity level. Under the same conditions, the strains harbouring this transporter accumulated more than 400 mM glycerol, whereas the glycerol/H+ symporter mutant presented less than 1 mM. Consistently, the strains able to accumulate glycerol survive 25-50% more than the stl1Δ mutant. Conclusions In this work, we report the contribution of the glycerol/H+ symporter Stl1p for the accumulation and maintenance of glycerol intracellular levels, and consequently cell survival at cold/near-freeze and freeze temperatures. These findings have a high biotechnological impact, as they show that any S. cerevisiae strain already in use can become more resistant to cold/freeze-thaw stress just by simply adding glycerol to the broth. The combination of low temperatures with extracellular glycerol will induce the transporter Stl1p. This solution avoids the use of transgenic strains, in particular in food industry.

  5. Serum-converted platelet lysate can substitute for fetal bovine serum in human mesenchymal stromal cell cultures.

    Science.gov (United States)

    Mojica-Henshaw, Mariluz P; Jacobson, Pam; Morris, Julie; Kelley, Linda; Pierce, Jan; Boyer, Michael; Reems, Jo-Anna

    2013-12-01

    Fetal bovine serum (FBS) is commonly used as a serum supplement for culturing human mesenchymal stromal cells (hMSCs). However, human cells grown in FBS, especially for extended periods, risk potential exposure to bovine immunogenic proteins and infectious agents. To address this issue, we investigated the ability of a novel human platelet serum supplement to substitute for FBS in hMSC cultures. Platelet lysate-serum (PL-serum) was converted from platelet lysate-plasma (PL-plasma) that was manufactured from pooled platelet-rich plasma (PRP) apheresis units. Growth factor levels and the number of residual intact platelets in PL-serum and PL-plasma were compared with enzyme-linked immunosorbent assays and flow cytometry, respectively. Proliferation responses of hMSCs cultured in PL-serum, PL-plasma, or FBS were assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, the immunophenotype of harvested hMSCs was evaluated by flow cytometry and tri-lineage differentiation potential was evaluated by assessing adipogenic, osteogenic and chondrogenic development. Selected growth factor levels in PL-serum were not significantly different from PL-plasma (P > 0.05). hMSC cultures supplemented with PL-serum had comparable growth kinetics to PL-plasma, and hMSC yields were consistently greater than with FBS. hMSCs harvested from cultures supplemented with PL-serum, PL-plasma or FBS had similar cell surface phenotypes and maintained tri-lineage differentiation potential. PL-serum, similar to PL-plasma, can substitute for FBS in hMSC cultures. Use of PL-serum, in contrast to PL-plasma, has an added advantage of not requiring addition of a xenogeneic source of heparin, providing a completely xeno-free culture medium. Copyright © 2013 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  6. Quality changes and freezing time prediction during freezing and thawing of ginger

    OpenAIRE

    Singha, Poonam; Muthukumarappan, Kasiviswanathan

    2015-01-01

    Abstract Effects of different freezing rates and four different thawing methods on chemical composition, microstructure, and color of ginger were investigated. Computer simulation for predicting the freezing time of cylindrical ginger for two different freezing methods (slow and fast) was done using ANSYS ? Multiphysics. Different freezing rates (slow and fast) and thawing methods significantly (P?

  7. Direct, Label-Free, and Rapid Transistor-Based Immunodetection in Whole Serum.

    Science.gov (United States)

    Gutiérrez-Sanz, Óscar; Andoy, Nesha M; Filipiak, Marcin S; Haustein, Natalie; Tarasov, Alexey

    2017-09-22

    Transistor-based biosensors fulfill many requirements posed upon transducers for future point-of-care diagnostic devices such as scalable fabrication and label-free and real-time quantification of chemical and biological species with high sensitivity. However, the short Debye screening length in physiological samples (<1 nm) has been a major drawback so far, preventing direct measurements in serum. In this work, we demonstrate how tailoring the sensing surface with short specific biological receptors and a polymer polyethylene glycol (PEG) can strongly enhance the sensor response. In addition, the sensor performance can be dramatically improved if the measurements are performed at elevated temperatures (37 °C instead of 21 °C). With this novel approach, highly sensitive and selective detection of a representative immunosensing parameter-human thyroid-stimulating hormone-is shown over a wide measuring range with subpicomolar detection limits in whole serum. To the best of our knowledge, this is the first demonstration of direct immunodetection in whole serum using transistor-based biosensors, without the need for sample pretreatment, labeling, or washing steps. The presented sensor is low-cost, can be easily integrated into portable diagnostics devices, and offers a competitive performance compared to state-of-the-art central laboratory analyzers.

  8. Changes of Serum Total and Free Testosterone Concentrations in Male Chronic Hemodialysis Patients with Secondary Hyperparathyroidism in Response to Cinacalcet Treatment

    Directory of Open Access Journals (Sweden)

    Piotr Kuczera

    2016-01-01

    Full Text Available Background/Aims: Calcium sensing receptor (CaSR is expressed, among others also in testis. Cinacalcet binds to the CaSR, increases sensitivity of CaSR to serum calcium and is used in the treatment of secondary hyperparathyroidism (sHPT in chronic hemodialysis patients (HDP. In most of male HDP, serum testosterone concentration is lower than in healthy males. The aim of this study was to assess the influence of six-month treatment with cinacalcet on the serum total and free testosterone concentration in male HDP with sHPT. Methods: 38 male, hemodialysed CKD patients with sHPT (PTH>300 pg/ml were enrolled into the study. In each patient serum PTH, total testosterone (TT and free testosterone (FT concentrations were assessed before the first dose of cinacalcet and then after 3 and 6 months of treatment. The results are presented as means with 95% confidence interval. Results: In 33 patients who completed the study cinacalcet treatment caused significant decrease of serum PTH from 1143 pg/ml (828 - 1458 pg/ml at the baseline, to 809 pg/ml (487 - 1132pg/ml after 3 month of treatment (p = 0.002, and to 607 pg/ml (281 - 934pg/ml; p Conclusion: Treatment with cinacalcet decreases serum total and free testosterone concentration in male hemodialysed patients with chronic kidney disease and secondary hyperparathyroidism.

  9. Synthetic surface for expansion of human mesenchymal stem cells in xeno-free, chemically defined culture conditions.

    Directory of Open Access Journals (Sweden)

    Paula J Dolley-Sonneville

    Full Text Available Human mesenchymal stem cells (HMSCS possess three properties of great interest for the development of cell therapies and tissue engineering: multilineage differentiation, immunomodulation, and production of trophic factors. Efficient ex vivo expansion of hMSCs is a challenging requirement for large scale production of clinical grade cells. Low-cost, robust, scalable culture methods using chemically defined materials need to be developed to address this need. This study describes the use of a xeno-free synthetic peptide acrylate surface, the Corning® Synthemax® Surface, for culture of hMSCs in serum-free, defined medium. Cell performance on the Corning Synthemax Surface was compared to cells cultured on biological extracellular matrix (ECM coatings in xeno-free defined medium and in traditional conditions on tissue culture treated (TCT plastic in fetal bovine serum (FBS supplemented medium. Our results show successful maintenance of hMSCs on Corning Synthemax Surface for eight passages, with cell expansion rate comparable to cells cultured on ECM and significantly higher than for cells in TCT/FBS condition. Importantly, on the Corning Synthemax Surface, cells maintained elongated, spindle-like morphology, typical hMSC marker profile and in vitro multilineage differentiation potential. We believe the Corning Synthemax Surface, in combination with defined media, provides a complete synthetic, xeno-free, cell culture system for scalable production of hMSCs.

  10. Self-supported ceramic substrates with directional porosity by mold freeze casting

    DEFF Research Database (Denmark)

    Gurauskis, Jonas; Graves, Christopher R.; Moreno, R.

    2016-01-01

    in a mold and applying directional freeze casting. Use of optimized suspension, cryoprotector additive and mold proved to deliver defect free ceramic films with high dimensional control. Microstructure analysis demonstrated the formation of desirable aligned porosity at macro-structural scale and resulted...... to be highly dependent on colloidal behaviour and freeze casting conditions. Manufactured green films were joined by lamination at room temperature and sintered to obtain symmetrical cells consisting of two porous self-supported substrate electrodes (∼420 μm) and dense yttria stabilized zirconia electrolyte...

  11. Freeze drying method

    International Nuclear Information System (INIS)

    Coppa, N.V.; Stewart, P.; Renzi, E.

    1999-01-01

    The present invention provides methods and apparatus for freeze drying in which a solution, which can be a radioactive salt dissolved within an acid, is frozen into a solid on vertical plates provided within a freeze drying chamber. The solid is sublimated into vapor and condensed in a cold condenser positioned above the freeze drying chamber and connected thereto by a conduit. The vertical positioning of the cold condenser relative to the freeze dryer helps to help prevent substances such as radioactive materials separated from the solution from contaminating the cold condenser. Additionally, the system can be charged with an inert gas to produce a down rush of gas into the freeze drying chamber to also help prevent such substances from contaminating the cold condenser

  12. Analysis of Serum Total and Free PSA Using Immunoaffinity Depletion Coupled to SRM: Correlation with Clinical Immunoassay Tests

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Tao; Hossain, Mahmud; Schepmoes, Athena A.; Fillmore, Thomas L.; Sokoll, Lori J.; Kronewitter, Scott R.; Izmirlian, Grant; Shi, Tujin; Qian, Weijun; Leach, Robin; Thompson, Ian M.; Chan, Daniel W.; Smith, Richard D.; Kagan, Jacob; Srinivastava, Sudhir; Rodland, Karin D.; Camp, David G.

    2012-08-03

    Sandwich immunoassay is the standard technique used in clinical labs for quantifying protein biomarkers for disease detection, monitoring and therapeutic intervention. Albeit highly sensitive, the development of a specific immunoassay is rather time-consuming and associated with extremely high cost due to the requirement for paired immunoaffinity reagents of high specificity. Recently, mass spectrometry-based methods, specifically selected reaction monitoring mass spectrometry (SRM-MS), have been increasingly applied to measure low abundance biomarker candidates in tissue and biofluids, owing to high sensitivity and specificity, simplicity of assay configuration, and great multiplexing capability. In this study, we report for the first time the development of immunoaffinity depletion-based workflows and SRM-MS assays that enable sensitive and accurate quantification of total and free prostate-specific antigen (PSA) in serum without the requirement for specific PSA antibodies. With stable isotope dilution and external calibration, low ng/mL level detection of both total and free PSA was consistently achieved in both PSA-spiked female serum samples and actual patient serum samples. Moreover, comparison of the results obtained when SRM PSA assays and conventional immunoassays were applied to the same samples showed very good correlation (R2 values ranging from 0.90 to 0.99) in several independent clinical serum sample sets, including a set of 33 samples assayed in a blinded test. These results demonstrate that the workflows and SRM assays developed here provide an attractive alternative for reliably measuring total and free PSA in human blood. Furthermore, simultaneous measurement of free and total PSA and many other biomarkers can be performed in a single analysis using high-resolution liquid chromatographic separation coupled with SRM-MS.

  13. To freeze or not to freeze embryos: clarity, confusion and conflict.

    Science.gov (United States)

    Goswami, Mohar; Murdoch, Alison P; Haimes, Erica

    2015-06-01

    Although embryo freezing is a routine clinical practice, there is little contemporary evidence on how couples make the decision to freeze their surplus embryos, or of their perceptions during that time. This article describes a qualitative study of 16 couples who have had in vitro fertilisation (IVF) treatment. The study question was 'What are the personal and social factors that patients consider when deciding whether to freeze embryos?' We show that while the desire for a baby is the dominant drive, couples' views revealed more nuanced and complex considerations in the decision-making process. It was clear that the desire to have a baby influenced couples' decision-making and that they saw freezing as 'part of the process'. However, there were confusions associated with the term 'freezing' related to concerns about the safety of the procedure. Despite being given written information, couples were confused about the practical aspects of embryo freezing, which suggests they were preoccupied with the immediate demands of IVF. Couples expressed ethical conflicts about freezing 'babies'. We hope the findings from this study will inform clinicians and assist them in providing support to couples confronted with this difficult decision-making.

  14. Supersonic free jet, molecular free regime

    International Nuclear Information System (INIS)

    Sanna, G.; Tomassetti, G.

    1999-01-01

    The structure of the free jet emitted by a converging nozzle as obtained by the method of characteristics by Ashkenas and Sherman is described in details. In particular the dependence of the field variable by the distance from the nozzle is given. The transition from continuum to molecular free regime is then considered and the sudden freeze approximation is introduced. The processing of monoatomic and polyatomic gasses is also considered [it

  15. Bulls grazing Kentucky 31 tall fescue exhibit impaired growth, semen quality, and decreased semen freezing potential

    Science.gov (United States)

    Serum prolactin (PRL) and testosterone concentrations, body weight, body composition, semen quality, and semen freezing potential for bulls grazing the toxic tall fescue (Lolium arundinaceum [Schreb.] Darbysh. ¼ Schedonorous arundinaceum [Schreb.] Dumort.) cultivar Kentucky 31 (E+) compared with a n...

  16. Serum-dependent effects of tamoxifen and cannabinoids upon C6 glioma cell viability.

    Science.gov (United States)

    Jacobsson, S O; Rongård, E; Stridh, M; Tiger, G; Fowler, C J

    2000-12-15

    In the present study, the effects of the combination of tamoxifen ((Z)-2[p-(1,2-diphenyl-1-butenyl)phenoxy]-N,N-dimethylamine citrate) and three cannabinoids (Delta(9)-tetrahydrocannabinol [Delta(9)-THC], cannabidiol, and anandamide [AEA]) upon the viability of C6 rat glioma cells was assessed at different incubation times and using different culturing concentrations of foetal bovine serum (FBS). Consistent with previous data for human glioblastoma cells, the tamoxifen sensitivity of the cells was increased as the FBS content of the culture medium was reduced from 10 to 0.4 and 0%. The cells expressed protein kinase C alpha and calmodulin (the concentration of which did not change significantly as the FBS concentration was reduced), but did not express estrogen receptors. Delta(9)-THC and cannabidiol, but not AEA, produced a modest reduction in cell viability after 6 days of incubation in serum-free medium, whereas no effects were seen in 10% FBS-containing medium. There was no observed synergy between the effects of tamoxifen and the cannabinoids upon cell viability.

  17. Effects of age and health on the euthyroid reference ranges for serum free thyroxine and free triiodothyronine

    International Nuclear Information System (INIS)

    Midgley, J.E.M.

    1985-01-01

    Age-related trends in serum free thyroxine (FT 4 ) and free triiodothyronine (FT 3 ) concentrations were measured in 7248 euthyroid subjects (age-range 3 months to 106 years). 5700 were patients referred to hospitals for investigation of suspected thyroid dysfunction, but who were diagnosed euthyroid. 1548 were healthy blood donors (age-range 18-63 years) with no indication of thyroid dysfunction. FT 4 concentrations were little affected by the age, the sex or the state of health of the subjects in either group. Serum FT 3 concentrations were significantly affected by both age and health factors. The upper limit of the euthyroid reference range for young subjects up to 15 years was about 20% higher (10.4 pmol/l) than for adult subjects older than 25 years (8.8 pmol/l). The change in the upper limits typical of young subjects to that typical of adults occurred steadily over the decade 15-25 years. After this age, little further change occurred, especially in healthy subjects. Additionally, the lower limit of the euthyroid range for FT 3 was extended by the inclusion in the reference group of patients referred to hospitals. Compared with the lower limit of the FT 3 range for healthy subjects (5 pmol/l), the corresponding limit for referred subjects (young or adult) was 3.5-3.8 pmol/l. Broadening of the FT 3 reference range was probably brought about by a significant number of patients in the hospital-referred group with the 'low-T 3 syndrome' of mild non-thyroidal illness. Accordingly, FT 3 was inferior to FT 4 in the discrimination of hypothyroidism, as FT 4 was unaffected by this phenomenon. Effects of age and non-thyroidal illness on serum FT 3 concentrations require great care when selecting subjects for a laboratory euthyroid reference range typical of the routine workload. Constraints on the choice of subjects for FT 4 reference ranges are less stringent. (orig.) [de

  18. Structure-property-processing correlations in freeze-cast composite scaffolds.

    Science.gov (United States)

    Hunger, Philipp M; Donius, Amalie E; Wegst, Ulrike G K

    2013-05-01

    Surprisingly few reports have been published, to date, on the structure-property-processing correlations observed in freeze-cast materials directionally solidified from polymer solutions, or ceramic or metal slurries. The studies that exist focus on properties of sintered ceramics, that is materials whose structure was altered by further processing. In this contribution, we report first results on correlations observed in alumina-chitosan-gelatin composites, which were chosen as a model system to test and compare the effect of particle size and processing parameters on their mechanical properties at a specific composition. Our study reveals that highly porous (>90%) hybrid materials can be manufactured by freeze casting, through the self-assembly of a polymer and a ceramic phase that occurs during directional solidification, without the need of additional processing steps such as sintering or infiltration. It further illustrates that the properties of freeze-cast hybrid materials can independently be tailored at two levels of their structural hierarchy, allowing for the simultaneous optimization of both mechanical and structural requirements. An increase in freezing rate resulted in decreases in lamellar spacing, cell wall thickness, pore aspect ratio and cross-sectional area, as well as increases in both Young's modulus and compressive yield strength. The mechanical properties of the composite scaffolds increased with an increasing particle size. The results show that both structure and mechanical properties of the freeze-cast composites can be custom-designed and that they are thus ideally suited for a large variety of applications that require high porosity at low or medium load-bearing capacity. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  19. Optimization of chemically defined cell culture media--replacing fetal bovine serum in mammalian in vitro methods

    DEFF Research Database (Denmark)

    van der Valk, J; Brunner, D; De Smet, K

    2010-01-01

    with an undefined and variable composition. Defined media supplements are commercially available for some cell types. However, information on the formulation by the companies is often limited and such supplements can therefore not be regarded as completely defined. The development of defined media is difficult......, reproducible and reduce the use of experimental animals. Good cell culture practice (GCCP) is an attempt to develop a common standard for in vitro methods. The implementation of the use of chemically defined media is part of the GCCP. This will decrease the dependence on animal serum, a supplement...... and often takes place in isolation. A workshop was organised in 2009 in Copenhagen to discuss strategies to improve the development and use of serum-free defined media. In this report, the results from the meeting are discussed and the formulation of a basic serum-free medium is suggested. Furthermore...

  20. [The value of serum free light chain in differential diagnosis of monoclonal gammopathy of renal significance].

    Science.gov (United States)

    Li, C; Wen, Y B; Li, H; Su, W; Li, J; Cai, J F; Chen, L M; Li, X M; Li, X W

    2017-08-08

    Objective: To investigate the value of serum free light chain (FLC) in differential diagnosis of monoclonal gammopathy of renal significance (MGRS). Methods: Forty-nine hospitalized patients who underwent renal biopsy in Peking Union Medical College Hospital between January 2013 and December 2015 were included. Monoclonal gammopathy was detected by serum protein electrophoresis (SPE), serum immunofixation electrophoresis (IFE), urine IFE and serum FLC. All patients were classified as MGRS ( n =32) and monoclonal gammopathy of undetermined significance (MGUS) ( n =17). Results: Renal lesions in MGRS subgroup included light chain amyloidosis ( n =24, 75.0%), light chain deposition disease ( n =7, 21.9%), and fibrillary glomerulopathy ( n =1, 3.1%). Renal diseases in MGUS subgroup included membranous nephropathy ( n =10), focal segmental glomerulosclerosi (FSGS) ( n =3), diabetic glomerulopathy ( n =1), Henoch-Schonlein purpura nephritis ( n =1), anti-GBM disease concurrent with membranous nephropathy ( n =1) and glomerulomegaly ( n =1). Positive number of SPE, serum IFE, urine IFE and abnormal number of serum FLC ratio in MGRS subgroup were 12, 16, 23 and 30, respectively. Positive number of SPE, serum IFE, urine IFE and abnormal number of serum FLC ratio in MGUS subgroup were 11, 17, 6 and 3, respectively. MGRS and MGUS subgroups differed significantly in positive rate of serum IFE ( P value for MGRS, which was helpful for differential diagnosis of patients who had contraindication to renal biopsy.

  1. Effect of radiation sterilization on free radicals content in dry beverages

    International Nuclear Information System (INIS)

    Hidayati, Nur; Sudiro, Sutjipto; Maha, Munsiah

    1998-01-01

    This study was done to determine free radicals content in several dry beverages, namely instant coffee, milk powder, cane sugar, nutrisari, tea, and cacao powder after radiation-sterilization at 25 kGy and stored at room (24 o C) and freezing (-13 o C) temperatures. Each sample was vacuum-packed in Al foil laminate pouches, then irradiated at 25 kGy in dry ice temperature. Free radicals contents in some samples after being dissolved or soaked in hot water (73 o C) then freeze-dried were also measured. The free radicals content was measured using Electron Spin Resonance (ESR) Spectroscopy and expressed in spectrum area/g in arbitrary unit. The results showed that free radicals content increased after irradiation, but it decreased immediately after storage especially at room temperature. After about 5 months storage either at room or freezing temperature, free radicals contents of irradiated coffee, milk, powder and cacao powder were similar to those of the unirradiated control, while in tea sample stored at freezing temperature, the free radicals content was still a little bit higher than that stored at room temperature which was already similar to the control. Free radicals content of irradiated sugar as well as nutrisari was higher than the other samples, and the radicals were relatively stable either at room or freezing temperatures. However, after dissolving or soaking the samples in water, the ESR spectra or free radicals contents of the irradiated samples were found to be identical to the control. This indicates that free radicals formed by irradiation in the samples disappear because termination. (authors)

  2. Serum free light chain reference values: a critical approach.

    Science.gov (United States)

    Altinier, Sara; Seguso, Mara; Zaninotto, Martina; Varagnolo, Mariacristina; Adami, Fausto; Angeli, Paolo; Plebani, Mario

    2013-05-01

    The clinical usefulness of serum free light chain (FLC) measurement in the management of patients with plasma cell proliferative disorders has been reported in several papers, and most clinical studies use the reference ranges declared by the manufacturer. The aim of the present study was to evaluate the reproducibility of FLCs immunoassay and to validate the reference range, before introducing it in routine setting. Internal quality control materials and a pool of fresh serum samples were used to evaluate imprecision; 162 fresh sera from healthy blood donors were analyzed to evaluate the reference range for FLCs. In order to verify the κ/λ FLC ratio, 43 sera from patients with polyclonal hypergammaglobulinemia were tested. The FLC immunoassay was performed using a nephelometer with the Freelite reagents. The imprecision studies performed using a serum pool tested with two different lots of reagents showed a mean CV of 16.09% for κFLC and of 16.72% for λFLC. Lower CV%s and different mean values were found by calculating the results from each specific lot separately, while different results were obtained using the control materials provided by the manufacturer. In reference subjects, the 2.5-97.5th percentiles were found to be 4.52-22.33 and 4.84-21.88mg/L for κFLC and λFLC, respectively. The range for κ/λ ratio (0.65-2.36) was validated with the values obtained from subjects with polyclonal hypergammaglobulinemia. In retesting 15 samples from blood donor subjects with a different lot of reagents, mean bias percentages of 17.60 for κFLC and 15.26 for λFLC were obtained. These findings confirm the lot-to-lot variability of the FLC assays also in the measurement of polyclonal light chains, as well as the need to carefully validate the reference values. Published by Elsevier Inc.

  3. Serum vitamin D levels in free-ranging koalas (Phascolarctos cinereus).

    Science.gov (United States)

    Pye, Geoffrey W; Ellis, William; FitzGibbon, Sean; Opitz, Brian; Keener, Laura; Hollis, Bruce W

    2013-06-01

    Due to climatic conditions in Northern America and Europe, koalas (Phascolarctos cinereus) are often housed indoors. Koala joeys raised in these environments are susceptible to the development of metabolic bone disease due to a lack of exposure to solar ultraviolet radiation to themselves and their dam. As an initial step toward describing vitamin D sufficiency and adequately measuring responses to supplementation, vitamin D values were calculated by using serum collected from 20 free-ranging koalas on St. Bees Island, Queensland, Australia. Vitamin D values ranged from 8.1 to 30.4 pg/ml (18.4 +/- 5.5 pg/ml) for 1, 25-hydroxyvitamin D, and from 1 to 14 nM/L (7.4 +/- 3.0 nM/L) for 25-hydroxyvitamin D. These koala serum vitamin D values are unusually low when compared with eutherian mammals. Although this study was limited in numbers and in the geographically range of the koalas sampled, it does suggest that the koala's requirement for vitamin D is low. Therefore, supplementation to prevent disease may be relatively easy to achieve because low doses will likely meet requirements. Caution should be taken to avoid intoxication if supplementing vitamin D in koalas.

  4. Spray-freeze-drying of nanosuspensions: the manufacture of insulin particles for needle-free ballistic powder delivery

    Science.gov (United States)

    Schiffter, Heiko; Condliffe, Jamie; Vonhoff, Sebastian

    2010-01-01

    The feasibility of preparing microparticles with high insulin loading suitable for needle-free ballistic drug delivery by spray-freeze-drying (SFD) was examined in this study. The aim was to manufacture dense, robust particles with a diameter of around 50 µm, a narrow size distribution and a high content of insulin. Atomization using ultrasound atomizers showed improved handling of small liquid quantities as well as narrower droplet size distributions over conventional two-fluid nozzle atomization. Insulin nanoparticles were produced by SFD from solutions with a low solid content (ballistic injection, the insulin nanoparticles were suspended in matrix formulations with a high excipient content (>300 mg ml−1) consisting of trehalose, mannitol, dextran (10 kDa) and dextran (150 kDa) (abbreviated to TMDD) in order to maximize particle robustness and density after SFD. With the increase in insulin content, the viscosity of the nanosuspensions increased. Liquid atomization was possible up to a maximum of 250 mg of nano-insulin suspended in a 1.0 g matrix. However, if a narrow size distribution with a good correlation between theoretical and measurable insulin content was desired, no more than 150 mg nano-insulin could be suspended per gram of matrix formulation. Particles were examined by laser light diffraction, scanning electron microscopy and tap density testing. Insulin stability was assessed using size exclusion chromatography (SEC), reverse phase chromatography and Fourier transform infrared (FTIR) spectroscopy. Densification of the particles could be achieved during primary drying if the product temperature (Tprod) exceeded the glass transition temperature of the freeze concentrate (Tg′) of −29.4°C for TMDD (3∶3∶3∶1) formulations. Particles showed a collapsed and wrinkled morphology owing to viscous flow of the freeze concentrate. With increasing insulin loading, the d (v, 0.5) of the SFD powders increased and particle size distributions got wider

  5. Trehalose in glycerol-free freezing extender enhances post-thaw survival of boar spermatozoa

    Science.gov (United States)

    ATHURUPANA, Rukmali; TAKAHASHI, Daisen; IOKI, Sumire; FUNAHASHI, Hiroaki

    2015-01-01

    Cryopreservation of boar semen is still considered suboptimal due to lower fertility as compared with fresh samples when glycerol, a permeating cryoprotectant, is used. Trehalose is a non-permeable cryoprotectant and nonreducing disaccharide known to stabilize proteins and biologic membranes. The aim of this study was to evaluate the cryosurvival and in vitro penetrability of boar spermatozoa when glycerol was replaced with trehalose in a freezing extender. Ejaculated Berkshire semen samples were diluted in egg yolk-based freezing extender containing glycerol (100 mM) or trehalose (0, 50, 100, 150, 200 and 250 mM) and cryopreserved using a straw freezing procedure. Thawed samples were analyzed for motility, viability, mitochondrial membrane potential (MMP), and acrosome integrity. In experiment 2, penetrability of spermatozoa cryopreserved with 100 mM glycerol or trehalose was examined. Replacement of cryoprotectant glycerol (100 mM) with trehalose had no effect on sperm viability, but replacing it with 100 mM trehalose improved motility, MMP and acrosome integrity significantly. Sperm motility and MMP were considerably higher in 100 mM trehalose, whereas the acrosome integrity was substantially higher in 100–250 mM trehalose. The in vitro penetration rate was also significantly higher in spermatozoa cryopreserved with trehalose (61.3%) than in those cryopreserved with glycerol (43.6%). In conclusion, 100 mM non-permeable trehalose can be used to replace glycerol, a permeating cryoprotectant, for maintenance of better post-thaw quality of boar spermatozoa. PMID:25754239

  6. [Cultivation of a permanent fish cell line in serum-free media special experiences with a cytotoxicity test for waste water samples

    Science.gov (United States)

    Kohlpoth, Martin; Rusche, Brigitte

    1997-01-01

    The use of fetal calf serum (FCS) as standard medium additive for the cell cultivation must be regarded critically from the point of view of animal welfare as well as for scientific reasons and makes it necessary to look for alternatives. In the last years an in vitro cytotoxicity assay for the testing of industrial waste waters with the permanent fish cell line RTG-2 was established and pre-validated as an alternative to the fish test with the golden orfe. The application of FCS is also a special problem with regard to the testing of waste waters in a cytotoxicity test so that FCS-alternatives were tested. The RTG-2 cells were successfully adapted to the two solvents Basal Medium Supplement (BMS) and Ultroser-G (U-G) that are used to replace serum. The characterisation of these adapted cell lines showed no significant differences in growth rate, adhesion rate, viability and sensitivity to chemicals in comparison to the original RTG-2 cells. On the determination of the cytotoxicity of industrial waste waters the RTG-2 cells adapted to the BMS medium indicated a clearly higher toxicity of the waste water samples than the original RTG-2 cells. This result confirms the thesis that serum components react with waste water elements and thus change the bio-availability of toxic compounds.

  7. Determination of free cisplatin in medium by differential pulse polarography after ultrasound and cisplatin treatment of a cancer cell culture

    International Nuclear Information System (INIS)

    Bernard, Vladan; Skorpikova, Jirina; Mornstein, Vojtech; Fojt, Lukas

    2011-01-01

    The in vitro study was carried out for detection of the cisplatin in free form and in culture medium, depending on various conditions of sonodynamic human ovarian cancer cells A2780 treatment by differential pulse polarography (DPP). For sonodynamic treatment, we used cisplatin alone and combined cisplatin/ultrasound treatments. The ultrasound exposure intensity of 1.0 and 2.0 Wcm 2 in far field for incubation periods 1, 24 and 48 h was used. The parameters of DPP measurements were - 1 s drop time, 5 mV.s -1 voltage scan rate, 50 mV modulation amplitude and negative scanning direction; platinum wire served as counter electrode and Ag|AgCl|3 M KCI as reference electrode. The results showed the dependence of free platinum quantities in culture medium on incubation time and treatment protocol. We found difference in concentration of free cisplatin between conventional application of cisplatin and sonodynamic treatment. The sonodynamic combined treatment of cisplatin and ultrasound field showed a higher cisplatin content in the culture medium than cisplatin treatment alone; a difference of 20% was observed for incubation time 48 h. The results also showed the influence of a time sequence of ultrasound and cytostatics in the sonodynamic treatment. The highest amount of free cisplatin in the solution was found for primary application of cisplatin and the subsequent ultrasound exposure. The quantity of free cisplatin increased with time, namely for time intervals 1-24 h. There was no difference between the DPP signal of cisplatin in reaction mixture containing cells in small quantities and micro-filtered mixture without cells. Thus, the DPP method is suitable for the detection and quantification of free cisplatin in the culture medium of cell suspension. Ultrasound field can be important factor during cytostatic therapy. (author)

  8. Effects of in-medium nucleon-nucleon cross sections on stopping observable and ratio of free protons in heavy-ion collisions at 400 MeV/nucleon

    Energy Technology Data Exchange (ETDEWEB)

    Su, Jun; Huang, Ching-Yuan [Sun Yat-sen University, Sino-French Institute of Nuclear Engineering and Technology, Zhuhai (China); Xie, Wen-Jie [Yuncheng University, Department of Physics, Yuncheng (China); Chinese Academy of Sciences, Institute of Modern Physics, Lanzhou (China); Zhang, Feng-Shou [Beijing Normal University, The Key Laboratory of Beam Technology and Material Modification of Ministry of Education, College of Nuclear Science and Technology, Beijing (China); Beijing Radiation Center, Beijing (China); National Laboratory of Heavy Ion Accelerator of Lanzhou, Center of Theoretical Nuclear Physics, Lanzhou (China)

    2016-07-15

    The effects of in-medium nucleon-nucleon cross sections on the stopping observable and ratio of free protons in heavy-ion collisions at 400 MeV/nucleon have been investigated within the framework of the IQMD+GEMINI model. Five kinds of in-medium corrections of nucleon-nucleon cross sections, which are considerably different in the referred energy and density, have been used in the model. It has been found that calculations of the stopping decrease when the in-medium cross sections decrease. Moreover, the ratio of free protons R{sub p} depends not only on the value of the in-medium factors but also on its isospin dependence. In order to investigate the isospin effect of in-medium factors on the ratio of free protons R{sub p}, the isospin dependence of in-medium factors has been adjusted and used in the model. The calculations have shown that the isospin dependence of in-medium factors does not impact the stopping, but impacts the ratio of free protons R{sub p}. When the in-medium factors relation f{sub nn}{sup med} > f{sub pp}{sup med} is used in the model, the calculated values of R{sub p} are larger than those in the f{sub nn}{sup med} < f{sub pp}{sup med} case. (orig.)

  9. Thermal stress state of cryogenic HP vessels under freezing and pressurization

    International Nuclear Information System (INIS)

    Tsybenko, A.S.; Kuranov, B.A.; Chepurnoj, A.D.; Shaposhnikov, V.A.; Krishchuk, N.G.

    1986-01-01

    A mathematical model is developed for thermomechanical processes in cryogenic HP vessels under freezing either by liquid and (or) gaseous cryogen and under pressurization. Equations of nonlinear nonstationary thermal conductivity and nonisothermal thermoelastoplasticity are used for the case of the theory off low with isotropic hardening. Semiempiricaldependences of nonstationary heat exchange for gaseous medium, experimental curves of cryogenic liquid boiling, mass exchange relationships are allowed for when formulating boundary conditions. The mathematical modelis realized on the basi of the finite element method in the form of highly automated program complex TERSOD (heat resistanceof vessels), oriented for computer of the Unified System. Heat and stress-strained states for three constructions of vessels are thoroughly studied under different conditions of gaseous, liquid and combined freezing with subsequent pressurization

  10. Does freeze-all policy affect IVF outcomes in poor responders?

    Science.gov (United States)

    Roque, Matheus; Valle, Marcello; Sampaio, Marcos; Geber, Selmo

    2017-12-27

    To evaluate whether the freeze-all strategy affects in vitro fertilization (IVF) outcomes in poor ovarian responders following the Bologna criteria. We performed a retrospective cohort study conducted between January 2012 and December 2016. A total of 433 poor responders (per the Bologna criteria) fulfilled the inclusion/exclusion criteria and were included in the study, with 277 patients included in the fresh group and 156 in the freeze-all group. All patients were submitted to controlled ovarian stimulation (COS) with a gonadotropin-releasing hormone antagonist protocol and cleavage-stage embryo transfer (ET). The main outcome measure was ongoing pregnancy rate. Secondary outcomes included implantation and clinical pregnancy rates. The freeze-all strategy was implemented when the progesterone serum level was >1.5 ng/mL on the trigger day, when the endometrium was <7 mm on the trigger day, or as a patient preference. Patients with previous failed fresh embryo transfer were also submitted to fresh or freeze-all strategy considering the indications mentioned above. The patients' mean age in the freeze-all group was 39.5±3.6 years, while that of patients in the fresh group was 39.7±3.8 years (P=0.54). The mean number of embryos transferred (nET) was 1.53±0.6 and 1.60±0.6 (P=0.12) in the freeze-all and fresh groups, respectively. Ongoing pregnancy rates did not significantly differ between the freeze-all and fresh groups (9.6% versus 10.1%, respectively; Relative Risk [RR]: 0.95; 95% CI: 0.52-1.73), nor did the clinical pregnancy rates (14.1% versus 13.7%, respectively; RR: 1.03; 95% CI: 0.63-1.76). Implantation rates were 9.6% and 9.8% (P=0.82) in the freeze-all and fresh groups, respectively. The logistic regression analysis (including age, antral follicle count [AFC], the number of retrieved oocytes, the number of mature oocytes, nET, and fresh versus freeze-all strategy) indicated that age (P<0.001) and the nET (P=0.039) were the only independent variables

  11. Effects of six substances on the growth and freeze-drying of Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Chen, He; Huang, Jie; Shi, Xiaoyu; Li, Yichao; Liu, Yu

    2017-01-01

    The efficacy of Lactobacillus delbrueckii subsp. bulgaricus as starter cultures for the dairy industry depends largely on the number of viable and active cells. Freeze-drying is the most convenient and successful method to preserve the bacterial cells. However, not all strains survived during freeze-drying. The effects of six substances including NaCl, sorbitol, mannitol, mannose, sodium glutamate, betaine added to the MRS medium on the growth and freeze-drying survival rate and viable counts of Lb. delbrueckii subsp. bulgaricus were studied through a single-factor test and Plackett-Burman design. Subsequently, the optimum freeze-drying conditions of Lb. delbrueckii subsp. bulgaricus were determined. Lb. delbrueckii subsp. bulgaricus survival rates were up to the maximum of 42.7%, 45.4%, 23.6%, while the concentrations of NaCl, sorbitol, sodium glutamate were 0.6%, 0.15%, 0.09%, respectively. In the optimum concentration, the viable counts in broth is 6.1, 6.9, 5.13 (×108 CFU/mL), respectively; the viable counts in freeze-drying power are 3.09, 5.2, 2.7 (×1010 CFU/g), respectively. Three antifreeze factors including NaCl, sorbitol, sodium glutamate have a positive effect on the growth and freeze-drying of Lb. delbrueckii subsp. bulgaricus. The results are beneficial for developing Lb. delbrueckii subsp. bulgaricus.

  12. Freezing during tapping tasks in patients with advanced Parkinson's disease and freezing of gait.

    Science.gov (United States)

    Delval, Arnaud; Defebvre, Luc; Tard, Céline

    2017-01-01

    Parkinson's disease patients with freezing of gait also experience sudden motor blocks (freezing) during other repetitive motor tasks. We assessed the proportion of patients with advanced PD and freezing of gait who also displayed segmental "freezing" in tapping tasks. Fifteen Parkinson's disease patients with freezing of gait were assessed. Freezing of gait was evaluated using a standardized gait trajectory with the usual triggers. Patients performed repetitive tapping movements (as described in the MDS-UPDRS task) with the hands or the feet in the presence or absence of a metronome set to 4 Hz. Movements were recorded with a video motion system. The primary endpoint was the occurrence of segmental freezing in these tapping tasks. The secondary endpoints were (i) the relationship between segmental episodic phenomena and FoG severity, and (ii) the reliability of the measurements. For the upper limbs, freezing was observed more frequently with a metronome (21% of trials) than without a metronome (5%). For the lower limbs, the incidence of freezing was higher than for the upper limbs, and was again observed more frequently in the presence of an auditory cue (47%) than in its absence (14%). Although freezing of the lower limbs was easily assessed during an MDS-UPDRS task with a metronome, it was not correlated with the severity of freezing of gait (as evaluated during a standardized gait trajectory). Only this latter was a reliable measurement in patients with advanced Parkinson's disease.

  13. Preparation and evaluation of freeze-dried Mag3 kits for 99m Tc-labelling

    International Nuclear Information System (INIS)

    El-Mohty, A.A.; El-Ghany, E.A.; El-Kolaly, M.T.; Raieh, M.; EL-Bary, A.A.

    1996-01-01

    The freeze-dried Mag 3 kits were designed for both ligand trans chelation and direct labelling techniques. The solution of Sn-Mag 3 was sterilized by 0.22 μU mill pore filtration and dispensed in a laminar flow hood (1 m I / vial) then, the vials were introduced to the lyophilized. The process of lyophilization was continued for 24 hours. At end of the cycle, the vials were closed under nitrogen. The moisture content of the freeze-dried Mag 3 kits was determined and it was found equal to 0.1% also, the losses of tin (II) during the freeze-drying cycle did not exceed 5%. It was found that the Mag 3 freeze-dried kits were sterile, pyrogen free and does not have any unexpected toxicity. The prepared Mag 3 freeze-dried kits have high radiochemical purity > 97% and high stability for more than 8 h after labelling. The biodistribution shows rapid renal excretion at 15 min post injection. 3 figs., 4 tabs

  14. The comparison of free androgen index and serum free testosterone levels in women with hirsutism or polycystic ovary syndrome

    Directory of Open Access Journals (Sweden)

    M. Metin Yıldırımkaya

    2011-06-01

    Full Text Available In many laboratories free testosterone can not be measured, so that free androgen index is suggested instead. The aim of this study was to compare free androgen index and serum free testosterone levels measured by radioimmunoassay in women with hirsutism or polycystic ovary syndrome.Materials and methods: Totally 94 women referred to the polyclinics of Ankara Numune Hospital were retrospectively included. Three patient groups were composed; 55 of hirsutism, 20 of polycystic ovary syndrome and 19 of both hirsutism and polycystic ovary syndrome. Total testosterone and sex hormone binding globuline levels were measured by chemiluminescence method and free testosterone levels were measured by radioimmunoassay. Free androgen index was calculated from total testosterone and sex hormone binding globuline.Results: There was a significant positive correlation between free testosterone and free androgen index in patients with hirsutism, in patients with polycystic ovary syndrome, in patients with hirsutism and polycystic ovary syndrome, and in total patient group [r(hirsutism=0,597, r(PCOS=0,617, r(hirsutism and PCOS=0,779, r(total patient group=0,649, P<0,01].Receiver operating characteristics curves were drawn to assess the diagnostic power of parameters for all patient groups [For hirsutism (n=55 auROC (FT=0,431 auROC (FAI=0,485] [For PCOS (n=20 auROC (FT=0,431 auROC (FAI=0,359] [For hirsutism and PCOS (n=19 auROC (FT=0,676 auROC (FAI=0,669]. In our study, free testosterone and free androgen index were found useful to diagnose ‘hirsutism and polycystic ovary syndrome’ but not others.Conclusion: Free androgen index can be used instead of free testosterone in hirsutism and polycystic ovary syndrome for diagnosis. J Clin Exp Invest 2011;2(2:152-6

  15. The association between use of metformin and change in serum CO_2 level after administration of contrast medium

    International Nuclear Information System (INIS)

    Kim, S.K.; Jung, J.; Jung, J.H.; Kim, K.Y.; Baek, J.-H.; Hahm, J.R.

    2016-01-01

    Aim: To evaluate the changes in serum creatinine and total CO_2 levels in patients receiving metformin during administration of contrast medium. Materials and methods: Patient records from January 2012 to December 2012 after the administration of contrast medium were reviewed retrospectively. A total of 924 patients were included for the final analysis. Of them, 105 received metformin during contrast medium administration, 112 were taking other oral hypoglycaemic agents, and 707 patients were not diabetic (controls). Results: No significant change in total CO_2 levels was detected (p=0.678). Metabolic acidosis was present in 33 (31.4%) metformin users, 31 (28.6%) other oral hypoglycaemic agent users, and 153 (21.6%) control patients. In the present logistic regression analysis, age, baseline levels of creatinine, and total CO_2 levels were associated with metabolic acidosis after contrast medium exposure. Conclusion: These data indicate the presence of a coexisting risk factor, other than metformin use, associated with metabolic acidosis after contrast medium exposure. No relationship was found between the use of metformin and metabolic acidosis during contrast medium exposure. - Highlights: • The use of metformin was not associated with metabolic acidosis after contrast exposure. • The coexisting risk factors for metabolic acidosis were present in patient with metabolic acidosis after contrast exposure. • There is a need to consider the maintenance of metformin during a CT scan in patients with a low risk for lactic acidosis.

  16. Effect of thyroxine on cellular oxygen-consumption and glucose uptake: evidence of an effect of total T4 and not "free T4"

    DEFF Research Database (Denmark)

    Kvetny, J; Matzen, L E

    1990-01-01

    Recent studies of cellular T4 and T3 uptake have indicated active transport of the hormones into the cell rather than passive diffusion of the non-protein bound fraction. In order to study the significance of the extracellular environment, oxygen consumption and glucose uptake were examined...... in human mononuclear blood cells. Cells were incubated in protein free medium and in human serum totally depleted of thyroid hormones by resin treatment and fixed amounts of T4 (total T4 = 0-50-100-5000 nmol/l; free T4 = 0-5-11-5600 pmol/l) were added. Thyroxine stimulated glucose uptake and oxygen......-consumption in a dose dependent manner but the T4 stimulation was dependent on the total concentration of T4 and did not differ between serum incubation or non-protein containing medium. Addition of ANS (100 mg/l) which inhibits binding of T4 to TBG, did not increase T4 effect in serum. Inhibition of the Na...

  17. Sperm DNA damage in relation to lipid peroxidation following freezing-thawing of boar semen

    OpenAIRE

    Fraser, L.; Strzeżek, J.; Wasilewska, K.; Pareek, C.S.

    2017-01-01

    This study investigated the relationships between lipid peroxidation (LPO) and sperm DNA damage following freezing-thawing of boar semen in different extenders. The comet assay was used to measure the extent of sperm DNA damage in a cryoprotectant-free extender or in cryoprotectant-based extenders after single and repeated freezing and thawing. As well as an analysis of sperm motion characteristics, mitochondrial function, membrane integrity, and lipid peroxidation (LPO) were assessed simulta...

  18. Using Power Ultrasound to Accelerate Food Freezing Processes: Effects on Freezing Efficiency and Food Microstructure.

    Science.gov (United States)

    Zhang, Peizhi; Zhu, Zhiwei; Sun, Da-Wen

    2018-05-31

    Freezing is an effective way of food preservation. However, traditional freezing methods have the disadvantages of low freezing efficiency and generation of large ice crystals, leading to possible damage of food quality. Power ultrasound assisted freezing as a novel technique can effectively reduce the adverse effects during freezing process. This paper gives an overview on recent researches of power ultrasound technique to accelerate the food freezing processes and illustrates the main principles of power ultrasound assisted freezing. The effects of power ultrasound on liquid food, model solid food as well as fruit and vegetables are discussed, respectively, from the aspects of increasing freezing rate and improving microstructure. It is shown that ultrasound assisted freezing can effectively improve the freezing efficiency and promote the formation of small and evenly distributed ice crystals, resulting in better food quality. Different inherent properties of food samples affect the effectiveness of ultrasound application and optimum ultrasound parameters depend on the nature of the samples. The application of ultrasound to the food industry is more likely on certain types of food products and more efforts are still needed to realize the industrial translation of laboratory results.

  19. Surface freezing of water

    OpenAIRE

    P?rez-D?az, J. L.; ?lvarez-Valenzuela, M. A.; Rodr?guez-Celis, F.

    2016-01-01

    Freezing, melting, evaporation and condensation of water are essential ingredients for climate and eventually life on Earth. In the present work, we show how surface freezing of supercooled water in an open container is conditioned and triggered?exclusively?by humidity in air. Additionally, a change of phase is demonstrated to be triggered on the water surface forming surface ice crystals prior to freezing of bulk. The symmetry of the surface crystal, as well as the freezing point, depend on ...

  20. Effect of culture medium on propagation and phenotype of corneal stroma-derived stem cells.

    Science.gov (United States)

    Sidney, Laura E; Branch, Matthew J; Dua, Harminder S; Hopkinson, Andrew

    2015-12-01

    The limbal area of the corneal stroma has been identified as a source of mesenchymal-like stem cells, which have potential for exploitation as a cell therapy. However, the optimal culture conditions are disputed and few direct media comparisons have been performed. In this report, we evaluated several media types to identify the optimal for inducing an in vitro stem cell phenotype. Primary human corneal stroma-derived stem cells (CSSCs) were extracted from corneoscleral rims. Culture in seven different media types was compared: Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS); M199 with 20% FBS; DMEM-F12 with 20% serum replacement, basic fibroblast growth factor and leukemia inhibitory factor (SCM); endothelial growth medium (EGM); semi-solid MethoCult; serum-free keratinocyte medium (K-SFM); and StemPro-34. Effects on proliferation, morphology, protein and messenger RNA expression were evaluated. All media supported proliferation of CSSCs with the exception of K-SFM and StemPro-34. Morphology differed between media: DMEM produced large cells, whereas EGM produced very small cells. Culture in M199 produced a typical mesenchymal stromal cell phenotype with high expression of CD105, CD90 and CD73 but not CD34. Culture in SCM produced a phenotype more reminiscent of a progenitor cell type with expression of CD34, ABCG2, SSEA-4 and PAX6. Culture medium can significantly influence CSSC phenotype. SCM produced a cell phenotype closest to that of a pluripotent stem cell, and we consider it to be the most appropriate for development as a clinical-grade medium for the production of CSSC phenotypes suitable for cell therapy. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  1. Serum Free Light Chains in Neoplastic Monoclonal Gammopathies: Relative Under-Detection of Lambda Dominant Kappa/Lambda Ratio, and Underproduction of Free Lambda Light Chains, as Compared to Kappa Light Chains, in Patients With Neoplastic Monoclonal Gammopathies.

    Science.gov (United States)

    Lee, Won Sok; Singh, Gurmukh

    2018-07-01

    Quantitative evaluation of serum free light chains is recommended for the work up of monoclonal gammopathies. Immunoglobulin light chains are generally produced in excess of heavy chains. In patients with monoclonal gammopathy, κ/λ ratio is abnormal less frequently with lambda chain lesions. This study was undertaken to ascertain if the levels of overproduction of the two light chain types and their detection rates are different in patients with neoplastic monoclonal gammopathies. Results of serum protein electrophoresis (SPEP), serum protein immunofixation electrophoresis (SIFE), urine protein electrophoresis (UPEP), urine protein immunofixation electrophoresis (UIFE), and serum free light chain assay (SFLCA) in patients with monoclonal gammopathies were examined retrospectively. The κ/λ ratios were appropriately abnormal more often in kappa chain lesions. Ratios of κ/λ were normal in about 25% of patients with lambda chain lesions in whom free homogenous lambda light chains were detectable in urine. An illustrative case suggests underproduction of free lambda light chains, in some instances. The lower prevalence of lambda dominant κ/λ ratio in lesions with lambda light chains is estimated to be due to relative under-detection of lambda dominant κ/λ ratio in about 25% of the patients and because lambda chains are not produced in as much excess of heavy chains as are kappa chains, in about 5% of the patients. The results question the medical necessity and clinical usefulness of the serum free light chain assay. UPEP/UIFE is under-utilized.

  2. The Effect of Calcusol™ to the Plasma Free Radical and Serum Creatinin in Mus Musculus Nephrolithiasis Model

    Directory of Open Access Journals (Sweden)

    A’liyatur Rosyidah

    2013-09-01

    Full Text Available Calcusol™ is a traditional medicine (jamu made from Tempuyung (Sonchus arvensis extract and is usually used for cure kidney stone disease. However, there has not been any studies which investigate the its mechanism. The aim of this study is to know the effect of Calcusol™ to the plasma free radical and serum creatinin of Mus musculus model for renal calcium-oxalate accumulation. This study is carried out by administration of Porang (Amorphophallus muelleri tuber flour for 3 months with the doses 6 mg/100g BW to induce renal calcium oxalate accumulation and Calcusol™ treatment for 7 days with the doses of 3.3mg/g BW. Group I was used as a control group. Group II was only given porang every day for 3 months. Group III was only given Calcusol™ for 7 days. Group IV were given porang for 3 months then given Calcusol™ for 7 days. Group V were given porang and Calcusol™ simultaneously for 3 months. Porang and CalcusolTM is administrated orally. Blood was collected from the tail of the animal for serum creatinin test and plasma free radical test using TBARS method. The data was analyzed using ANOVA followed by Tukey HSD to compare the means employing SPSS 16.0 for windows. The result of the research shows that the treatment Calcusol™ on mice model for renal calcium oxalate accumulation. The content of MDA at group I, group II, group III, group IV and group V, respectivelyis 0.81±0.5 mg/ml; 2.63±0.8 mg/ml; 0.56±0.5 mg/ml; 2.09±0.9 mg/ml and 0.17±0.17 mg/ml. The content of serum creatinine are 0.15±0.04 mg/dL; 0.13±0.03 mg/dL; 0.12±0.08 mg/dL; 0.11±0.016 mg/dL and 0.14±0.015 mg/dL at group I, group II, group III, Group IV, and group V respectively. This indicates that Calcusol™ decreases plasma free radical production during renal stone formation, while serum creatinin reduced but not significantly changed. Calcusol™ has an effect to decrease free radical during renalstone formation inmice model for calcium oxalate accumulation

  3. A comparison of freezing-damage during isochoric and isobaric freezing of the potato

    OpenAIRE

    Lyu, Chenang; Nastase, Gabriel; Ukpai, Gideon; Serban, Alexandru; Rubinsky, Boris

    2017-01-01

    Background Freezing is commonly used for food preservation. It is usually done under constant atmospheric pressure (isobaric). While extending the life of the produce, isobaric freezing has detrimental effects. It causes loss of food weight and changes in food quality. Using thermodynamic analysis, we have developed a theoretical model of the process of freezing in a constant volume system (isochoric). The mathematical model suggests that the detrimental effects associated with isobaric freez...

  4. The influence of parenteral nitrogen feeding on free amino acid composition of blood serum and hepatic tissue of irradiated rats

    International Nuclear Information System (INIS)

    Mil'ko, V.I.; Kirichenko, A.V.; Chalaya, L.A.

    1985-01-01

    A considerable change in the free am ino acid composition of blood serum and hepatic tissued was noted on the 7th and 14th days following total-body X-irradiation of rats with a dose of 2.9 Gy. The total free amino acid content of blood serum increased and that of hepatic tissue decreased by 85% (on an average) as compared to the intact controls. Quantitative changes in the content of individual amino acids were analysed. Polyamine injected enterally for 7 days and parenterally for 3 days after irradiation a the elimination of the postirradiation changes in the amino acid balance

  5. Serum concentrations and effects of detomidine delivered orally to horses in three different mediums.

    Science.gov (United States)

    Ramsay, Edward C; Geiser, Dennis; Carter, Wyndee; Tobin, Thomas

    2002-10-01

    To compare the effect of orally delivered detomidine on head posture when administered alone or in combination with two different food items, and to determine the serum concentrations of detomidine after oral delivery. Prospective randomized experimental study. Fifteen adult grade mares weighing 328-537 kg. The horses were randomly assigned to one of the three treatment groups (five horses each). The groups were given detomidine (0.06 mg kg -1 ): alone; mixed with 3 mL of an apple sauce and gum mixture; or mixed with 3 mL molasses. Head droop, measured before treatment and at 15, 30, 45, 60, 75, 90, and 105 minutes after treatment, was used to evaluate sedation. Yohimbine (0.1 mg kg -1 IV) was administered after the 90-minute evaluation. Blood samples were collected from the detomidine-alone group before treatment and at 15, 30, 45, 60, 75, and 90 minutes after treatment. Sera were analyzed for detomidine equivalent concentrations by an ELISA. Head droop percentages were compared using a repeated measures analysis of variance. Significant mean head droop developed in each treatment group by 30 minutes and persisted until reversal with yohimbine. After yohimbine administration, head positions returned to 87-91% of pre-treatment levels. There were no significant differences among the oral treatment groups at any time. Mean serum detomidine equivalents increased slowly until 45-minute post-administration, but never exceeded 30 ng mL -1 . Orally administered detomidine results in measurable serum drug concentrations using any of the delivery mediums investigated, and can be expected to produce profound head droop in horses approximately 45 minutes after administration. Copyright © 2002 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.

  6. Shift of microRNA profile upon glioma cell migration using patient-derived spheroids and serum-free conditions

    DEFF Research Database (Denmark)

    Munthe, Sune; Halle, Bo; Boldt, Henning B

    2017-01-01

    Glioblastoma multiforme (GBM) is the most frequent malignant primary brain tumor. A major reason for the overall median survival being only 14.6 months is migrating tumor cells left behind after surgery. Another major reason is tumor cells having a so-called cancer stem cell phenotype being...... therefore resistant towards traditional chemo- and radiotherapy. A group of novel molecular targets are microRNAs (miRNAs). MiRNAs are small non-coding RNAs exerting post-transcriptional regulation of gene expression. The aim of this study was to identify differentially expressed miRNAs in migrating GBM...... cells using serum-free stem cell conditions. We used patient-derived GBM spheroid cultures for a novel serum-free migration assay. MiRNA expression of migrating tumor cells isolated at maximum migration speed was compared with corresponding spheroids using an OpenArray Real-Time PCR System. The mi...

  7. Micro-bead injection spectroscopy for label-free automated determination of immunoglobulin G in human serum.

    Science.gov (United States)

    Ramos, Inês I; Magalhães, Luís M; Barreiros, Luisa; Reis, Salette; Lima, José L F C; Segundo, Marcela A

    2018-01-01

    Immunoglobulin G (IgG) represents the major fraction of antibodies in healthy adult human serum, and deviations from physiological levels are a generic marker of disease corresponding to different pathologies. Therefore, screening methods for IgG evaluation are a valuable aid to diagnostics. The present work proposes a rapid, automatic, and miniaturized method based on UV-vis micro-bead injection spectroscopy (μ-BIS) for the real-time determination of human serum IgG with label-free detection. Relying on attachment of IgG in rec-protein G immobilized in Sepharose 4B, a bioaffinity column is automatically assembled, where IgG is selectively retained and determined by on-column optical density measurement. A "dilution-and-shoot" approach (50 to 200 times) was implemented without further sample treatment because interferences were flushed out of the column upon sample loading, with minimization of carryover and cross-contamination by automatically discarding the sorbent (0.2 mg) after each determination. No interference from human serum albumin at 60 mg mL -1 in undiluted sample was found. The method allowed IgG determination in the range 100-300 μg mL -1 (corresponding to 5.0-60 mg mL -1 in undiluted samples), with a detection limit of 33 μg mL -1 (1.7 mg mL -1 for samples, dilution factor of 50). RSD values were time-to-result decreased from several hours to times, showing the potential of the proposed approach as a point-of-care method. Graphical abstract Micro-Bead Injection Spectroscopy method for real time, automated and label-free determination of total serum human Immunoglobulin G (IgG). The method was designed for Lab-on-Valve (LOV) platforms using a miniaturised protein G bioaffinity separative approach. IgG are separated from serum matrix components upon quantification with low non-specific binding in less than 5 min.

  8. Warm and cold fermionic dark matter via freeze-in

    International Nuclear Information System (INIS)

    Klasen, Michael; Yaguna, Carlos E.

    2013-01-01

    The freeze-in mechanism of dark matter production provides a simple and intriguing alternative to the WIMP paradigm. In this paper, we analyze whether freeze-in can be used to account for the dark matter in the so-called singlet fermionic model. In it, the SM is extended with only two additional fields, a singlet scalar that mixes with the Higgs boson, and the dark matter particle, a fermion assumed to be odd under a Z 2 symmetry. After numerically studying the generation of dark matter, we analyze the dependence of the relic density with respect to all the free parameters of the model. These results are then used to obtain the regions of the parameter space that are compatible with the dark matter constraint. We demonstrate that the observed dark matter abundance can be explained via freeze-in over a wide range of masses extending down to the keV range. As a result, warm and cold dark matter can be obtained in this model. It is also possible to have dark matter masses well above the unitarity bound for WIMPs

  9. Surface freezing of water.

    Science.gov (United States)

    Pérez-Díaz, J L; Álvarez-Valenzuela, M A; Rodríguez-Celis, F

    2016-01-01

    Freezing, melting, evaporation and condensation of water are essential ingredients for climate and eventually life on Earth. In the present work, we show how surface freezing of supercooled water in an open container is conditioned and triggered-exclusively-by humidity in air. Additionally, a change of phase is demonstrated to be triggered on the water surface forming surface ice crystals prior to freezing of bulk. The symmetry of the surface crystal, as well as the freezing point, depend on humidity, presenting at least three different types of surface crystals. Humidity triggers surface freezing as soon as it overpasses a defined value for a given temperature, generating a plurality of nucleation nodes. An evidence of simultaneous nucleation of surface ice crystals is also provided.

  10. Changes during hibernation in different phospholipid and free and esterified cholesterol serum levels in black bears

    Science.gov (United States)

    Chauhan, V.; Sheikh, A.; Chauhan, A.; Tsiouris, J.; Malik, M.; Vaughan, M.

    2002-01-01

    During hibernation, fat is known to be the preferred source of energy. A detailed analysis of different phospholipids, as well as free and esterified cholesterol, was conducted to investigate lipid abnormalities during hibernation. The levels of total phospholipids and total cholesterol in the serum of black bears were found to increase significantly in hibernation as compared with the active state. Both free and esterified cholesterol were increased in the hibernating state in comparison with the active state (P biologie mole??culaire. All rights reserved.

  11. Nitrogen deposition may enhance soil carbon storage via change of soil respiration dynamic during a spring freeze-thaw cycle period.

    Science.gov (United States)

    Yan, Guoyong; Xing, Yajuan; Xu, Lijian; Wang, Jianyu; Meng, Wei; Wang, Qinggui; Yu, Jinghua; Zhang, Zhi; Wang, Zhidong; Jiang, Siling; Liu, Boqi; Han, Shijie

    2016-06-30

    As crucial terrestrial ecosystems, temperate forests play an important role in global soil carbon dioxide flux, and this process can be sensitive to atmospheric nitrogen deposition. It is often reported that the nitrogen addition induces a change in soil carbon dioxide emission in growing season. However, the important effects of interactions between nitrogen deposition and the freeze-thaw-cycle have never been investigated. Here we show nitrogen deposition delays spikes of soil respiration and weaken soil respiration. We found the nitrogen addition, time and nitrogen addition×time exerted the negative impact on the soil respiration of spring freeze-thaw periods due to delay of spikes and inhibition of soil respiration (p nitrogen), 39% (medium-nitrogen) and 36% (high-nitrogen) compared with the control. And the decrease values of soil respiration under medium- and high-nitrogen treatments during spring freeze-thaw-cycle period in temperate forest would be approximately equivalent to 1% of global annual C emissions. Therefore, we show interactions between nitrogen deposition and freeze-thaw-cycle in temperate forest ecosystems are important to predict global carbon emissions and sequestrations. We anticipate our finding to be a starting point for more sophisticated prediction of soil respirations in temperate forests ecosystems.

  12. Defined xenogeneic-free and hypoxic environment provides superior conditions for long-term expansion of human adipose-derived stem cells.

    Science.gov (United States)

    Yang, Sufang; Pilgaard, Linda; Chase, Lucas G; Boucher, Shayne; Vemuri, Mohan C; Fink, Trine; Zachar, Vladimir

    2012-08-01

    Development and implementation of therapeutic protocols based on stem cells or tissue-engineered products relies on methods that enable the production of substantial numbers of cells while complying with stringent quality and safety demands. In the current study, we aimed to assess the benefits of maintaining cultures of adipose-derived stem cells (ASCs) in a defined culture system devoid of xenogeneic components (xeno-free) and hypoxia over a 49-day growth period. Our data provide evidence that conditions involving StemPro mesenchymal stem cells serum-free medium (SFM) Xeno-Free and hypoxia (5% oxygen concentration) in the culture atmosphere provide a superior proliferation rate compared to a standard growth environment comprised of alpha-modified Eagle medium (A-MEM) supplemented with fetal calf serum (FCS) and ambient air (20% oxygen concentration) or that of A-MEM supplemented with FCS and hypoxia. Furthermore, a flow cytometric analysis and in vitro differentiation assays confirmed the immunophenotype stability and maintained multipotency of ASCs when expanded under xeno-free conditions and hypoxia. In conclusion, our data demonstrate that growth conditions utilizing a xeno-free and hypoxic environment not only provide an improved environment for the expansion of ASCs, but also set the stage as a culture system with the potential broad spectrum utility for regenerative medicine and tissue engineering applications.

  13. Mean free paths and in-medium scattering cross sections of energetic nucleons in neutron-rich nucleonic matter within the relativistic impulse approximation

    International Nuclear Information System (INIS)

    Jiang Weizhou; Li Baoan; Chen Liewen

    2007-01-01

    The mean free paths and in-medium scattering cross sections of energetic nucleons in neutron-rich nucleonic matter are investigated using the nucleon optical potential obtained within the relativistic impulse approximation with the empirical nucleon-nucleon scattering amplitudes and the nuclear densities obtained in the relativistic mean-field model. It is found that the isospin-splitting of nucleon mean free paths, sensitive to the imaginary part of the symmetry potential, changes its sign at certain high kinetic energy. The in-medium nucleon-nucleon cross sections are analytically and numerically demonstrated to be essentially independent of the isospin asymmetry of the medium and increase linearly with density in the high-energy region where the relativistic impulse approximation is applicable

  14. Diagnostic significance of the serum thyroid hormone indicies in various thyroid diseases

    International Nuclear Information System (INIS)

    Han, B.H.; Ko, S.M.; Yoon, S.R.; Ro, H.K.

    1980-01-01

    In an attempt to evaluate the diagnostic significance of the serum thyroid hormones in various thyroid function states, the author measured serum T 3 uptake, serum T 3 , serum T 4 , serum free T 4 and free T 4 index in 27 cases of normal subjects, 11 cases of hypothyroidism, 152 cases of euthyroidism and 81 cases of hyperthyroidism by the radioimmunoassay method. The results were as follows: 1) The ranges of serum thyroid hormones in normal subjects were, serum T 3 uptake; 27.4-42.1%, serum T 3 ; 93-245 ng/dl, serum T 4 ; 4.08-12.9 ng/dl and serum free T 4 ; 0.57-1.53 ng/dl (M+-2 S.D.). 2) Free T 4 index and serum T 4 show relatively high diagnostic value in euthyroidism group, and serum T 3 and T 4 in hypothyroidism group, while serum T 3 , free T 4 and T 4 show relatively high diagnostic value in hyperthyroidism group. 3) There were significant correlation between free T 4 index and serum T 4 (r=0.68) and between free T 4 index and serum free T 4 (r=0.67) in hyperthyroidism group. (author)

  15. Differentiating Graves' disease from subacute thyroiditis using ratio of serum free triiodothyronine to free thyroxine.

    Science.gov (United States)

    Sriphrapradang, Chutintorn; Bhasipol, Adikan

    2016-09-01

    The measurement of free thyroid hormone, instead of the total form, is more commonly used in current practice. We aimed to evaluate the usefulness of the ratio of serum free triiodothyronine (FT3, pg/mL) to free thyroxine (FT4, ng/dL) for differentiating Graves' disease from subacute thyroiditis. Medical records of thyrotoxic patients aged >15 years who had measurement of FT3, FT4 and thyrotropin on the first diagnosis of thyrotoxicosis before initiating treatment were retrospectively reviewed. Data were collected from all clinics, and were not limited to the endocrine clinic. Pregnant women were excluded. A total of 548 patients (468 with Graves' disease, 40 with subacute thyroiditis and 40 with toxic adenoma/multinodular goiter) were recruited. Mean age was 43.9 ± 15.4 years. Most were female 434 (79.2%), and goiter was present in 55.3%. Prevalence of T3-toxicosis and T4-toxicosis were 5.6% and 6.6%, respectively. Mean FT3/FT4 ratios were 4.62 ± 2 (10(-2) pg/ng) in patients with Graves' disease and 2.73 ± 0.5 in subacute thyroiditis. The area under the ROC curve of the FT3/FT4 ratio for diagnosis of Graves' disease was 0.83 (95%CI, 0.76-0.91). Cutoff level of this ratio >4.4 offered sensitivity of 47.2% and specificity of 92.8%. FT3/FT4 ratio of >4.4 (10(-2) pg/ng) may help in differentiating the cause of thyrotoxicosis.

  16. Weights, hematology and serum chemistry of seven species of free-ranging tropical pelagic seabirds

    Science.gov (United States)

    Work, Thierry M.

    1996-01-01

    I established reference values for weight, hematology, and serum chemistry for seven species of free-ranging Hawaiian tropical pelagic seabirds comprising three orders (Procellariiformes, Pelecaniformes, Charadriiformes) and six families (Procellariidae, Phaethontidae, Diomedeidae, Sulidae, Fregatidae, and Laridae). Species examined included 84 Hawaiian dark-rumped petrels (Pterodoma phaeopygia), 90 wedge-tailed shearwaters (Puffinus pacificus), 151 Laysan albatrosses (Diomedea immutabilis), 69 red-footed boobies (Sula sula), 154 red-tailed tropicbirds (Phaeton rubricauda), 90 great frigatebirds (Fregata minor), and 72 sooty terns (Sterna fuscata). Hematocrit, total plasma solids, total and differential white cell counts, serum glucose, calcium, phosphorus, uric acid, total protein, albumin, globulin, aspartate aminotransferase and creatinine phosphokinase were analyzed. Among and within species, hematology and chemistry values varied with age, sex, season, and island of collection. Despite this variation, order-wide trends were observed.

  17. Fragmentation, dissipative expansion, and freeze-out in medium energy heavy-ion collisions

    International Nuclear Information System (INIS)

    Gross, D.H.E.; Li Baoan; DeAngelis, A.R.

    1992-01-01

    The collision dynamics of 96 Mo + 96 Mo at 55 A MeV is simulated by solving numerically the Boltzmann-Uehling-Uhlenbeck (BUU) transport equation for the one-body phase-space distribution-function of nucleons with and without Coulomb interaction. A scatter-plot of the one-body density distribution shows an initial compression, subsequent homogeneous expansion, a breaking into ''fragments'', a very slow creeping expansion up to a freeze-out and in the case of included Coulomb-interaction a Coulomb-explosion. In the calculation which included Coulomb-interaction the overall shape of the ensemble of dense fragments is spherical. The fragments are created over the entire volume of the dense part of the source and not at the surface only. In the simulation without Coulomb interaction a doughnut-like shape may develop. (orig.)

  18. The influence of freezing rates on bovine pericardium tissue Freeze-drying

    Directory of Open Access Journals (Sweden)

    Camila Figueiredo Borgognoni

    2009-12-01

    Full Text Available The bovine pericardium has been used as biomaterial in developing bioprostheses. Freeze-drying is a drying process that could be used for heart valve's preservation. The maintenance of the characteristics of the biomaterial is important for a good heart valve performance. This paper describes the initial step in the development of a bovine pericardium tissue freeze-drying to be used in heart valves. Freeze-drying involves three steps: freezing, primary drying and secondary drying. The freezing step influences the ice crystal size and, consequently, the primary and secondary drying stages. The aim of this work was to investigate the influence of freezing rates on the bovine pericardium tissue freeze-drying parameters. The glass transition temperature and the structural behaviour of the lyophilized tissues were determined as also primary and secondary drying time. The slow freezing with thermal treatment presented better results than the other freeze-drying protocols.O pericárdio bovino é um material utilizado na fabricação de biopróteses. A liofilização é um método de secagem que vem sendo estudado para a conservação de válvulas cardíacas. A preservação das características do biomaterial é de fundamental importância no bom funcionamento das válvulas. Este artigo é a primeira etapa do desenvolvimento do ciclo de liofilização do pericárdio bovino. Liofilização é o processo de secagem no qual a água é removida do material congelado por sublimação e desorção da água incongelável, sob pressão reduzida. O congelamento influencia o tamanho do cristal de gelo e, consequentemente, a secagem primária e secundária. O objetivo deste estudo foi verificar a influência das taxas de congelamento nos parâmetros de liofilização do pericárdio bovino. Determinou-se a temperatura de transição vítrea e o comportamento estrutural do pericárdio bovino liofilizado. Determinou-se o tempo da secagem primária e secundária. O

  19. Two-step freezing of hybridoma cells in 96-well microculture plates.

    Science.gov (United States)

    Pĕknicová, J; Kristofová, H

    1985-01-01

    Stabile hybridoma cells, colonies of hybridoma cells 14 days after fusion of immune spleen and myeloma cells, myeloma cells and fibroblasts cultured in 96-well microculture plates were frozen by the method of two-step freezing. The culture medium was aspirated, and 50 microliter of the medium containing a cryoprotectant (5% dimethyl sulphoxide) was added for 10 min at room temperature. The plates were put into microtene bags, placed at -25 degrees C in a freezer for 30 min and then stored at -100 degrees C in liquid nitrogen vapour. Plates with cells were thawed rapidly in a 50 degree C water bath. After thawing the hybrid cells were viable and continued to produce the specific antibody.

  20. Diverse Requirements for Microglial Survival, Specification, and Function Revealed by Defined-Medium Cultures.

    Science.gov (United States)

    Bohlen, Christopher J; Bennett, F Chris; Tucker, Andrew F; Collins, Hannah Y; Mulinyawe, Sara B; Barres, Ben A

    2017-05-17

    Microglia, the resident macrophages of the CNS, engage in various CNS-specific functions that are critical for development and health. To better study microglia and the properties that distinguish them from other tissue macrophage populations, we have optimized serum-free culture conditions to permit robust survival of highly ramified adult microglia under defined-medium conditions. We find that astrocyte-derived factors prevent microglial death ex vivo and that this activity results from three primary components, CSF-1/IL-34, TGF-β2, and cholesterol. Using microglial cultures that have never been exposed to serum, we demonstrate a dramatic and lasting change in phagocytic capacity after serum exposure. Finally, we find that mature microglia rapidly lose signature gene expression after isolation, and that this loss can be reversed by engrafting cells back into an intact CNS environment. These data indicate that the specialized gene expression profile of mature microglia requires continuous instructive signaling from the intact CNS. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Interrelationships of serum testosterone and free testosterone index with FFM and strength in aging men.

    Science.gov (United States)

    Roy, Tracey Ann; Blackman, Marc R; Harman, S Mitchell; Tobin, Jordan D; Schrager, Matthew; Metter, E Jeffery

    2002-08-01

    Muscle mass and strength losses during aging may be associated with declining levels of serum testosterone (T) in men. Few studies have shown a direct relationship between T and muscle mass and strength. Subjects were 262 men, aged 24-90 yr, from the Baltimore Longitudinal Study of Aging, who had T and sex hormone-binding globulin sex hormone-binding globulin (SHBG) measurements, from which the free T index (FTI) was calculated (T/SHBG) from serum samples collected longitudinally since 1963, total body fat mass and arm and leg fat-free mass (FFM) by dual-energy X-ray absorptiometry and arm and leg strength by dynanomometry. Mixed-effects models estimated T and FTI at the time of mass and strength measurements. Age, total body fat, arm and leg FFM, T, and FTI were significantly associated with concentric and eccentric strength. FTI, not T, was modestly, but directly, related to arm and leg strength after fat, arm and leg FFM, height, and age were accounted for and indirectly through body mass. FTI is a better predictor of arm and leg strength than T in aging men.

  2. Effect of storage duration on cytokine stability in human serum and plasma.

    Science.gov (United States)

    Vincent, Fabien B; Nim, Hieu T; Lee, Jacinta P W; Morand, Eric F; Harris, James

    2018-06-14

    Quantification of analytes such as cytokines in serum samples is intrinsic to translational research in immune diseases. Optimising pre-analytical conditions is critical for ensuring study quality, including evaluation of cytokine stability. We aimed to evaluate the effect on cytokine stability of storage duration prior to freezing of serum, and compare to plasma samples obtained from patients with systemic lupus erythematosus (SLE). Protein stability was analysed by simultaneously quantifying 18 analytes using a custom multi-analyte profile in SLE patient serum and plasma samples that had been prospectively stored at 4 °C for pre-determined periods between 0 and 30 days, prior to freezing. Six analytes were excluded from analysis, because most tested samples were above or below the limit of detection. Amongst the 12 analysed proteins, 11 did not show significant signal degradation. Significant signal degradation was observed from the fourth day of storage for a single analyte, CCL19. Proteins levels were more stable in unseparated serum compared to plasma for most analytes, with the exception of IL-37 which appears slightly more stable in plasma. Based on this, a maximum 3 days of storage at 4 °C for unseparated serum samples is recommended for biobanked samples intended for cytokine analysis in studies of human immune disease. Copyright © 2018 Elsevier Ltd. All rights reserved.

  3. Pre-cure freezing affects proteolysis in dry-cured hams.

    Science.gov (United States)

    Bañón, S; Cayuela, J M; Granados, M V; Garrido, M D

    1999-01-01

    Several parameters (sodium chloride, moisture, intramuscular fat, total nitrogen, non-protein nitrogen, white precipitates, free tyrosine, L* a* b* values and acceptability) related with proteolysis during the curing were compared in dry-cured hams manufactured from refrigerated and frozen/thawed raw material. Pre-cure freezing increased the proteolysis levels significantly (pcured meat, although it does not significantly affect the sensory quality of the dry-cured ham.

  4. Adaptive and freeze-tolerant heteronetwork organohydrogels with enhanced mechanical stability over a wide temperature range

    Science.gov (United States)

    Gao, Hainan; Zhao, Ziguang; Cai, Yudong; Zhou, Jiajia; Hua, Wenda; Chen, Lie; Wang, Li; Zhang, Jianqi; Han, Dong; Liu, Mingjie; Jiang, Lei

    2017-06-01

    Many biological organisms with exceptional freezing tolerance can resist the damages to cells from extra-/intracellular ice crystals and thus maintain their mechanical stability at subzero temperatures. Inspired by the freezing tolerance mechanisms found in nature, here we report a strategy of combining hydrophilic/oleophilic heteronetworks to produce self-adaptive, freeze-tolerant and mechanically stable organohydrogels. The organohydrogels can simultaneously use water and oil as a dispersion medium, and quickly switch between hydrogel- and organogel-like behaviours in response to the nature of the surrounding phase. Accordingly, their surfaces display unusual adaptive dual superlyophobic in oil/water system (that is, they are superhydrophobic under oil and superoleophobic under water). Moreover, the organogel component can inhibit the ice crystallization of the hydrogel component, thus enhancing the mechanical stability of organohydrogel over a wide temperature range (-78 to 80 °C). The organohydrogels may have promising applications in complex and harsh environments.

  5. Nutritional manipulation of primate retinas, I: effects of lutein or zeaxanthin supplements on serum and macular pigment in xanthophyll-free rhesus monkeys.

    Science.gov (United States)

    Neuringer, Martha; Sandstrom, Marita M; Johnson, Elizabeth J; Snodderly, D Max

    2004-09-01

    The xanthophylls lutein (L) and zeaxanthin (Z) are the primary components of macular pigment (MP) and may protect the macula from age-related degeneration (AMD). In this study, L or Z was fed to rhesus monkeys reared on xanthophyll-free diets to follow the accumulation of serum carotenoids and MP over time. Eighteen rhesus monkeys were fed xanthophyll-free semipurified diets from birth until 7 to 16 years. The diets of six were then supplemented with pure L and six with pure Z at 3.9 micromol/kg per day (2.2 mg/kg per day) for 24 to 56 weeks. At baseline and 4- to 12-week intervals during supplementation, serum carotenoids were measured by HPLC, and MP density was estimated by two-wavelength reflectometry. Serum carotenoids and MP were also measured in monkeys fed a stock diet. Monkeys fed xanthophyll-free diets had no L or Z in serum and no detectable MP. During supplementation, serum L or Z increased rapidly over the first 4 weeks and from 16 weeks onward maintained similar levels, both several times higher than in stock-diet-fed monkeys. The central peak of MP optical density increased to a relatively steady level by 24 to 32 weeks in both L- and Z-fed groups. Rhesus monkeys fed a stock diet had lower blood concentrations of L than those found in humans and other nonhuman primates. Rhesus monkeys respond to either dietary L or Z supplementation with increases in serum xanthophylls and MP, even after life-long xanthophyll deficiency. These animals provide a potential model to study mechanisms of protection from AMD. Copyright Association for Research in Vision and Ophthalmology

  6. Freezing of liquid alkali metals as screened ionic plasmas

    International Nuclear Information System (INIS)

    Badirkhan, Z.; Rovere, M.; Tosi, M.P.

    1990-08-01

    The relationship between Wigner crystallization of the classical ionic plasma and the liquid-solid transition of alkali metals is examined within the density wave theory of freezing. Freezing of the classical plasma on a rigid neutralizing background into the bcc structure is first re-evaluated, in view of recent progress in the determination of its thermodynamic functions by simulation and of the known difficulties of the theory relating to the order parameter at the (200) star of reciprocal lattice vectors. Freezing into the fcc structure is also considered in this context and found to be unfavoured. On allowing for long-wavelength deformability of the background, the ensuing appearance of a volume change on freezing into the bcc structure is accompanied by reduced stability of the fluid phase and by an increase in the entropy of melting. Freezing of alkali metals into the bcc structure is next evaluated, taking their ionic pair structure as that of an ionic plasma reference fluid screened by conduction electrons and asking that the correct ionic coupling strength at liquid-solid coexistence should be approximately reproduced. The ensuring values of the volume and entropy changes across the phase transition, as estimated from the theory by two alternative routes, are in reasonable agreement with experiment. The order parameters of the phase transition, excepting the (200) one, conform rather closely to a Gaussian behaviour and yield a Lindemann ratio in reasonable agreement with the empirical value for melting of bcc crystals. It is suggested that ionic ordering at the (200) star in the metal may be (i) assisted by medium range ordering in the conduction electrons, as indicated by differences in X-ray and neutron diffraction intensities from the liquid, and/or (ii) quite small in the hot bcc solid. Such a possible premelting behaviour of bcc metals should be worth testing experimentally by diffraction. (author). 48 refs, 1 fig., 1 tab

  7. Serum complexed and free prostate-specific antigen (PSA) for the diagnosis of the polycystic ovarian syndrome (PCOS).

    Science.gov (United States)

    Diamandis, Eleftherios P; Stanczyk, Frank Z; Wheeler, Sarah; Mathew, Anu; Stengelin, Martin; Nikolenko, Galina; Glezer, Eli N; Brown, Marshall D; Zheng, Yingye; Chen, Yen-Hao; Wu, Hsiao-Li; Azziz, Ricardo

    2017-10-26

    Polycystic ovarian syndrome (PCOS) is a common cause of reproductive and metabolic dysfunction. We hypothesized that serum prostate-specific antigen (PSA) may constitute a new biomarker for hyperandrogenism in PCOS. We conducted a cross-sectional study of 45 women with PCOS and 40 controls. Serum from these women was analyzed for androgenic steroids and for complexed PSA (cPSA) and free PSA (fPSA) with a novel fifth- generation assay with a sensitivity of ~10 fg/mL for cPSA and 140 fg/mL for fPSA. cPSA and fPSA levels were about three times higher in PCOS compared to controls. However, in PCOS, cPSA and fPSA did not differ according to waist-to-hip ratio, Ferriman-Gallwey score, or degree of hyperandrogenemia or oligo-ovulation. In PCOS and control women, serum cPSA and fPSA levels were highly correlated with each other, and with free and total testosterone levels, but not with other hormones. Adjusting for age, body mass index (BMI) and race, cPSA was significantly associated with PCOS, with an odds ratio (OR) of 5.67 (95% confidence interval [CI]: 1.86, 22.0). The OR of PCOS for fPSA was 7.04 (95% CI: 1.65, 40.4). A multivariate model that included age, BMI, race and cPSA yielded an area-under-the-receiver-operating-characteristic curve of 0.89. Serum cPSA and fPSA are novel biomarkers for hyperandrogenism in PCOS and may have value for disease diagnosis.

  8. Objective video quality assessment method for freeze distortion based on freeze aggregation

    Science.gov (United States)

    Watanabe, Keishiro; Okamoto, Jun; Kurita, Takaaki

    2006-01-01

    With the development of the broadband network, video communications such as videophone, video distribution, and IPTV services are beginning to become common. In order to provide these services appropriately, we must manage them based on subjective video quality, in addition to designing a network system based on it. Currently, subjective quality assessment is the main method used to quantify video quality. However, it is time-consuming and expensive. Therefore, we need an objective quality assessment technology that can estimate video quality from video characteristics effectively. Video degradation can be categorized into two types: spatial and temporal. Objective quality assessment methods for spatial degradation have been studied extensively, but methods for temporal degradation have hardly been examined even though it occurs frequently due to network degradation and has a large impact on subjective quality. In this paper, we propose an objective quality assessment method for temporal degradation. Our approach is to aggregate multiple freeze distortions into an equivalent freeze distortion and then derive the objective video quality from the equivalent freeze distortion. Specifically, our method considers the total length of all freeze distortions in a video sequence as the length of the equivalent single freeze distortion. In addition, we propose a method using the perceptual characteristics of short freeze distortions. We verified that our method can estimate the objective video quality well within the deviation of subjective video quality.

  9. SLAPex Freeze/Thaw 2015: The First Dedicated Soil Freeze/Thaw Airborne Campaign

    Science.gov (United States)

    Kim, Edward; Wu, Albert; DeMarco, Eugenia; Powers, Jarrett; Berg, Aaron; Rowlandson, Tracy; Freeman, Jacqueline; Gottfried, Kurt; Toose, Peter; Roy, Alexandre; hide

    2016-01-01

    Soil freezing and thawing is an important process in the terrestrial water, energy, and carbon cycles, marking the change between two very different hydraulic, thermal, and biological regimes. NASA's Soil Moisture Active/Passive (SMAP) mission includes a binary freeze/thaw data product. While there have been ground-based remote sensing field measurements observing soil freeze/thaw at the point scale, and airborne campaigns that observed some frozen soil areas (e.g., BOREAS), the recently-completed SLAPex Freeze/Thaw (F/T) campaign is the first airborne campaign dedicated solely to observing frozen/thawed soil with both passive and active microwave sensors and dedicated ground truth, in order to enable detailed process-level exploration of the remote sensing signatures and in situ soil conditions. SLAPex F/T utilized the Scanning L-band Active/Passive (SLAP) instrument, an airborne simulator of SMAP developed at NASA's Goddard Space Flight Center, and was conducted near Winnipeg, Manitoba, Canada, in October/November, 2015. Future soil moisture missions are also expected to include soil freeze/thaw products, and the loss of the radar on SMAP means that airborne radar-radiometer observations like those that SLAP provides are unique assets for freeze/thaw algorithm development. This paper will present an overview of SLAPex F/T, including descriptions of the site, airborne and ground-based remote sensing, ground truth, as well as preliminary results.

  10. Quantification of Maternal Serum Cell-Free Fetal DNA in Early-Onset Preeclampsia

    Directory of Open Access Journals (Sweden)

    Mulan Ren

    2013-04-01

    Full Text Available The aim of this study was to determine whether the increased serum cell-free fetal DNA (cffDNA level of gravidas developed into early-onset preeclampsia (EOPE subsequently in the early second trimesters is related to prenatal screening markers. Serum was collected from 1011 gravidas. The level of cffDNA and prenatal screening markers were analyzed in 20 cases with EOPE and 20 controls. All fetuses were male. The maternal serum cffDNA level was assessed by amplification of the Y chromosome specific gene. Correlations between the variables were examined. (Logged cffDNA in EOPE (median, 3.08; interquartile range, 2.93–3.68 was higher than controls (median, 1.79; interquartile range, 1.46–2.53. The increased level of (logged cffDNA was correlated significantly with the increased human chorionic gonadotropin (HCG level (r = 0.628, p < 0.001. Significant reciprocal correlations between cffDNA and babies’ birth weight as well as gestation weeks at delivery were noted (r = −0.516, p = 0.001; r = −0.623, p < 0.001, respectively. The sensitivity and specificity of cffDNA to discriminate between the EOPE cases and the controls were 90% and 85%, respectively. CffDNA is a potential marker for EOPE, which had a significant reciprocal correlation with babies’ birth weight and gestation weeks at delivery. Moreover, it may help in indicating the underlying hypoxic condition in the placenta.

  11. Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium

    Directory of Open Access Journals (Sweden)

    Denise Salzig

    2016-01-01

    Full Text Available The manufacture of human mesenchymal stem cells (hMSCs for clinical applications requires an appropriate growth surface and an optimized, preferably chemically defined medium (CDM for expansion. We investigated a new protein/peptide-free CDM that supports the adhesion, growth, and detachment of an immortalized hMSC line (hMSC-TERT as well as primary cells derived from bone marrow (bm-hMSCs and adipose tissue (ad-hMSCs. We observed the rapid attachment and spreading of hMSC-TERT cells and ad-hMSCs in CDM concomitant with the expression of integrin and actin fibers. Cell spreading was promoted by coating the growth surface with collagen type IV and fibronectin. The growth of hMSC-TERT cells was similar in CDM and serum-containing medium whereas the lag phase of bm-hMSCs was prolonged in CDM. FGF-2 or surface coating with collagen type IV promoted the growth of bm-hMSCs, but laminin had no effect. All three cell types retained their trilineage differentiation capability in CDM and were detached by several enzymes (but not collagenase in the case of hMSC-TERT cells. The medium and coating did not affect detachment efficiency but influenced cell survival after detachment. CDM combined with cell-specific surface coatings and/or FGF-2 supplements is therefore as effective as serum-containing medium for the manufacture of different hMSC types.

  12. Serum thyroid stimulating hormone, total and free T4 during the neonatal period: Establishing regional reference intervals

    Directory of Open Access Journals (Sweden)

    Sara Sheikhbahaei

    2014-01-01

    Full Text Available Context: Congenital hypothyroidism (CH, the most common etiology of preventable mental retardation in children, is estimated to be more prevalent among Asian population. Aims: Since thyroid function tests (TFTs varied among different ages and geographical regions, in this study, the neonatal thyroid reference intervals in a healthy neonatal population is determined for the first time in Iran. Settings and Design: A cross-sectional study performed on 246 healthy term newborns aged between 2 days and 1 month. Materials and Methods: Blood samples were obtained by venipuncture from all subjects. The median, 2.5 th , 5 th , 95 th , and 97.5 th percentile of serum thyroid-stimulating hormone (TSH, as well as the total and free T4 were assessed among different age groups. Statistical Analysis Used: Predictive Analytics Software (PASW Statistics 18 was used for the analysis. Results: Serum TSH, total and free T4 concentration peaked in 5 th to 7 th days of life, continued over 2 weeks, then decreased and started reaching to adult reference range. A significant negative correlation between age and serum concentration of TSH (P = 0.02, total T4 (P = 0.01 and free T4 (P = 0.01 was found. Conclusion: This study yielded fairly different values for TFTs compared compared values found in other countries and also different from values reported for laboratory kits we used. These differences were assumed to be due to variations in ethnicity, age, and laboratory methods used. Due to the lack of international standardization, conducting multicenter studies helps in making a more precise evaluation of thyroid status in neonates.

  13. Effects of various freezing containers for vitrification freezing on mouse oogenesis.

    Science.gov (United States)

    Kim, Ji Chul; Kim, Jae Myeoung; Seo, Byoung Boo

    2016-01-01

    In the present study, various freezing containers were tested for mouse embryos of respective developmental stages; embryos were vitrified and then their survival rate and developmental rate were monitored. Mouse two cell, 8 cell, and blastula stage embryos underwent vitrification freezing-thawing and then their recovery rate, survival rate, development rate, and hatching rate were investigated. EM-grid, OPS, and cryo-loop were utilized for vitrification freezing-thawing of mouse embryos. It was found that recovery rate and survival rate were higher in the group of cryo-loop compared to those of EM-grid (p containers on vitrified embryos of respective developmental stages; it was demonstrated that higher developmental rate was shown in more progressed (or developed) embryos with more blastomeres. There was however, no difference in embryonic development rate was shown amongst containers. Taken together, further additional studies are warranted with regards to 1) manipulation techniques of embryos for various vitrification freezing containers and 2) preventive measures against contamination via liquid nitrogen.

  14. Effects of different rearing and feeding systems on lipid oxidation and antioxidant capacity of freeze-dried egg yolks.

    Science.gov (United States)

    Pignoli, Giovanni; Rodriguez-Estrada, Maria Teresa; Mandrioli, Mara; Barbanti, Lorenzo; Rizzi, Laura; Lercker, Giovanni

    2009-12-23

    Lipid oxidation and antioxidant capacity of freeze-dried egg yolks produced with two rearing systems (battery cages and free-range) and two types of feedings (conventional and organic) were studied. Nine fresh egg yolks of each crossed treatment were pooled, frozen for a month, freeze-dried, vacuum-packed, and kept at -18 degrees C until analysis. No significant differences were observed in the lipid (58.0-62.1%) and total sterol contents (33.0-35.5 g/kg of lipids) of the freeze-dried egg yolks. Free rearing and conventional feeding systems resulted in significantly higher total tocopherol, alpha-tocopherol, and lutein contents, as compared to the battery cage and the organic feed, respectively. However, no significant differences were found in lipid oxidation (peroxide value = 0.7-0.9 mequiv of O(2)/kg of fat; thiobarbituric reactive substances = 1.0-1.3 mg of malonylaldehyde/kg of sample) and cholesterol oxidation (28.8-43.5 mg of cholesterol oxidation products/kg of lipids; 0.08-0.12% oxidized cholesterol) of freeze-dried egg yolks except for 7alpha-hydroxycholesterol, which was significantly lower in samples obtained with organic feed.

  15. Physical degradation of membrane electrode assemblies undergoing freeze/thaw cycling: Micro-structure effects

    Energy Technology Data Exchange (ETDEWEB)

    Kim, S. [Fuel Cell Dynamics and Diagnostics Laboratory, Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, University Park, PA 16802 (United States); Research and Development Division, Hyundai Motor Company, Yongin 446-912 (Korea); Mench, M.M. [Fuel Cell Dynamics and Diagnostics Laboratory, Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, University Park, PA 16802 (United States)

    2007-11-22

    The objective of this work is to investigate physical damage of polymer electrolyte fuel cell (PEFC) materials subjected to freeze/thaw cycling. Effects of membrane electrode assembly micro-structures (catalyst layer cracking, membrane thickness, and membrane reinforcement) and diffusion media with micro-porous layers were analyzed by comparing scanning electron microscopy images of freeze/thaw cycled samples (-40 C/70 C) with those of virgin material and thermal cycled samples without freezing (5 C/70 C). Ex situ testing performed in this study has revealed a strong direction for the material choices in the PEFC and confirmed the previous computational model in the literature [S. He, M.M. Mench, J. Electrochem. Soc., 153 (2006) A1724-A1731; S. He, S.H. Kim, M.M. Mench, J. Electrochem. Soc., in press]. Specifically, the membrane electrode assemblies were found to be a source of water that can damage the catalyst layers under freeze/thaw conditions. Damage was found to occur almost exclusively under the channel, and not under the land (the graphite that touches the diffusion media). Conceptually, the best material to mitigate freeze-damage is a crack free virgin catalyst layer on a reinforced membrane that is as thin as possible, protected by a stiff diffusion media. (author)

  16. Plasma and serum lipidomics of healthy white adults shows characteristic profiles by subjects' gender and age.

    Directory of Open Access Journals (Sweden)

    Masaki Ishikawa

    Full Text Available Blood is a commonly used biofluid for biomarker discovery. Although blood lipid metabolites are considered to be potential biomarker candidates, their fundamental properties are not well characterized. We aimed to (1 investigate the matrix type (serum vs. plasma that may be preferable for lipid biomarker exploration, (2 elucidate age- and gender-associated differences in lipid metabolite levels, and (3 examine the stability of lipid metabolites in matrix samples subjected to repeated freeze-thaw cycles. Using liquid chromatography-mass spectrometry, we performed lipidomic analyses for fasting plasma and serum samples for four groups (15 subjects/group of young and elderly (25-34 and 55-64 years old, respectively males and females and for an additional aliquot of samples from young males, which were subjected to repeated freeze-thaw cycles. Lysophosphatidylcholine and diacylglycerol levels were higher in serum than in plasma samples, suggesting that the clotting process influences serum lipid metabolite levels. Gender-associated differences highlighted that the levels of many sphingomyelin species were significantly higher in females than in males, irrespective of age and matrix (plasma and serum. Age-associated differences were more prominent in females than in males, and in both matrices, levels of many triacylglycerols were significantly higher in elderly females than in young females. Plasma and serum levels of most lipid metabolites were reduced by freeze-thawing. Our results indicate that plasma is an optimal matrix for exploring lipid biomarkers because it represents the original properties of an individual's blood sample. In addition, the levels of some blood lipid species of healthy adults showed gender- and age-associated differences; thus, this should be considered during biomarker exploration and its application in diagnostics. Our fundamental findings on sample selection and handling procedures for measuring blood lipid metabolites

  17. The value of a freeze

    International Nuclear Information System (INIS)

    Bethe, H.A.; Long, F.A.

    1988-01-01

    This paper reports on the rapid increase in public support for a nuclear-freeze agreement---that is, a mutual freeze on the testing, production and further deployment of nuclear weapons---which has been a remarkable political phenomenon. In less than a year, support has grown from a few volunteers collecting signatures on petitions to a congressional vote in which supporters of a freeze very nearly prevailed. This fall, eight states and the District of Columbia will vote on freeze referendums. Already Wisconsin voters have overwhelmingly voted yes in such a referendum. There are many reasons for this strong support for a freeze, including fear of nuclear war, resistance to high levels of military spending and opposition to particular military policies of the Reagan administration. But to most supporters, the chief purpose of a freeze is simple: it is to help stop an immense, continuing, dangerous and incredibly costly arms race between the two superpowers

  18. The Freezing Bomb

    Science.gov (United States)

    Mills, Allan

    2010-01-01

    The extreme pressures that are generated when water freezes were traditionally demonstrated by sealing a small volume in a massive cast iron "bomb" and then surrounding it with a freezing mixture of ice and salt. This vessel would dramatically fail by brittle fracture, but no quantitative measurement of bursting pressure was available. Calculation…

  19. Fundamental Technical Elements of Freeze-fracture/Freeze-etch in Biological Electron Microscopy

    Science.gov (United States)

    Freeze-fracture/freeze-etch describes a process whereby specimens, typically biological or nanomaterial in nature, are frozen, fractured, and replicated to generate a carbon/platinum "cast" intended for examination by transmission electron microscopy. Specimens are subjected to u...

  20. Calorimetric and spectroscopic properties of small globular proteins (bovine serum albumin, hemoglobin) after free radical generation

    International Nuclear Information System (INIS)

    Farkas, N.; Belagyi, J.; Lorinczy, D.

    2003-01-01

    Mild oxidation of -SH-containing proteins (serum albumin, hemoglobin) by Ce(IV)-ions in the presence of the spin trap phenyl-tert-butylnitrone (PBN) resulted in the appearance of strongly immobilized nitroxide free radicals which evidences the formation of thiyl radicals on the thiol site of the proteins. In hydroxyl free radical generating system a fraction of strongly immobilized nitroxide radicals was also detected in these proteins, which implies that the oxidation of a fraction of the thiol groups was also involved in the free radical reaction. According to the differential scanning calorimetry (DSC) experiments the melting processes of the proteins were calorimetrically irreversible, therefore the two-state kinetic model was used to evaluate the experiments. The results support the view that site-specific interaction of SH-containing proteins with hydroxyl and thiyl free radicals is able to modify the internal dynamics of proteins and affect the conformation of large molecules

  1. Impregnation of leather during "freeze-drying"

    DEFF Research Database (Denmark)

    Storch, Mikkel; Vestergaard Poulsen Sommer, Dorte; Hovmand, Ida

    2016-01-01

    Freeze-drying is a recognized method for the preservation of waterlogged objects. Naturally, freeze-drying has also been used for waterlogged archaeological leather often after treatment with Na2.EDTA and impregnation with PEG; but the treated leather sometimes suffers from “excessive drying......” becoming too stiff and brittle. The aim of this study was to examine the effect of a conventional freeze-drying method against an alternative freeze-drying method that preserves the natural moisture content of the leather. Both new and archaeological waterlogged leather were included in the study...... suggest that the process which takes place within the leather during the freeze-drying in not actual freeze-drying, but rather a sophisticated way of distributing the impregnating agent. The pure ice phase freezes out, but the impregnating agent remains liquid as the temperature does not become low enough...

  2. Building the Method to Determine the Rate of Freezing Water in Penaeus monodon of the Freezing Process

    OpenAIRE

    Nguyen Tan Dzung; Trinh Van Dzung; Tran Duc Ba

    2012-01-01

    The method of determination the rate of freezing water in Penaeus monodon of freezing process was established on base the equation of energy balance in warming up process Penaeus monodon after freezing to determine specific heat of Penaeus monodon. The result obtained was built the mathematical model (19) to determine the rate of freezing water according to the freezing temperature of Penaeus monodon. The results indicated that when water was completely frozen (ω = 1 or 100%), the optimal fre...

  3. Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture

    Science.gov (United States)

    Pijuan-Galitó, Sara; Tamm, Christoffer; Schuster, Jens; Sobol, Maria; Forsberg, Lars; Merry, Catherine L. R.; Annerén, Cecilia

    2016-01-01

    Reliable, scalable and time-efficient culture methods are required to fully realize the clinical and industrial applications of human pluripotent stem (hPS) cells. Here we present a completely defined, xeno-free medium that supports long-term propagation of hPS cells on uncoated tissue culture plastic. The medium consists of the Essential 8 (E8) formulation supplemented with inter-α-inhibitor (IαI), a human serum-derived protein, recently demonstrated to activate key pluripotency pathways in mouse PS cells. IαI efficiently induces attachment and long-term growth of both embryonic and induced hPS cell lines when added as a soluble protein to the medium at seeding. IαI supplementation efficiently supports adaptation of feeder-dependent hPS cells to xeno-free conditions, clonal growth as well as single-cell survival in the absence of Rho-associated kinase inhibitor (ROCKi). This time-efficient and simplified culture method paves the way for large-scale, high-throughput hPS cell culture, and will be valuable for both basic research and commercial applications. PMID:27405751

  4. Advantages of liquid nitrogen freezing of Penaeus monodon over conventional plate freezing

    OpenAIRE

    Chakrabarti, R.; Chaudhury, D.R.

    1987-01-01

    Liquid nitrogen frozen products are biochemically and organoleptically superior to conventional plate frozen products but beneficial effect of liquid nitrogen freezing over conventional plate freezing can exist only up to 59 days at a commercial storage temperature of -18°C.

  5. The relationship between serum total testosterone and free testosterone levels with serum hemoglobin and hematocrit levels: a study in 1221 men.

    Science.gov (United States)

    Shin, Yu Seob; You, Jae Hyung; Cha, Jai Seong; Park, Jong Kwan

    2016-12-01

    To investigate the relationship between serum total testosterone (TT) and free testosterone (FT) levels in men with anemia. We reviewed the records of 1221 subjects between March 2009 and December 2014. All the subjects' blood samples were drawn for TT and FT assays. Their serum hemoglobin (Hb) and serum hematocrit (Hct) levels were measured. The primary objective of our study was to investigate the association between TT and FT levels with Hb and Hct levels. The mean age was 59.82 ± 12.71 years. The mean TT and FT levels were 4.54 ± 2.02 ng/mL and 10.63 ± 3.69 pg/mL, respectively. The mean Hb and Hct levels were 14.72 ± 1.34 g/dL and 43.11 ± 3.75%, respectively. Subjects with low TT (<2.35 ng/mL) had low Hb and Hct levels (p < 0.001, p < 0.001, respectively). TT was positively associated with FT, Hb, and Hct. TT and FT levels were significantly lower in older men. Subjects with low TT and FT levels had low Hb and Hct levels. This suggests that TT and FT play a significant role in erythropoiesis. Testosterone replacement therapy may be effective in men with hypogonadism to reduce the incidence of anemia.

  6. Transcriptional and metabolomic analysis of Ascophyllum nodosum mediated freezing tolerance in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Nair Prasanth

    2012-11-01

    Full Text Available Abstract Background We have previously shown that lipophilic components (LPC of the brown seaweed Ascophyllum nodosum (ANE improved freezing tolerance in Arabidopsis thaliana. However, the mechanism(s of this induced freezing stress tolerance is largely unknown. Here, we investigated LPC induced changes in the transcriptome and metabolome of A. thaliana undergoing freezing stress. Results Gene expression studies revealed that the accumulation of proline was mediated by an increase in the expression of the proline synthesis genes P5CS1 and P5CS2 and a marginal reduction in the expression of the proline dehydrogenase (ProDH gene. Moreover, LPC application significantly increased the concentration of total soluble sugars in the cytosol in response to freezing stress. Arabidopsis sfr4 mutant plants, defective in the accumulation of free sugars, treated with LPC, exhibited freezing sensitivity similar to that of untreated controls. The 1H NMR metabolite profile of LPC-treated Arabidopsis plants exposed to freezing stress revealed a spectrum dominated by chemical shifts (δ representing soluble sugars, sugar alcohols, organic acids and lipophilic components like fatty acids, as compared to control plants. Additionally, 2D NMR spectra suggested an increase in the degree of unsaturation of fatty acids in LPC treated plants under freezing stress. These results were supported by global transcriptome analysis. Transcriptome analysis revealed that LPC treatment altered the expression of 1113 genes (5% in comparison with untreated plants. A total of 463 genes (2% were up regulated while 650 genes (3% were down regulated. Conclusion Taken together, the results of the experiments presented in this paper provide evidence to support LPC mediated freezing tolerance enhancement through a combination of the priming of plants for the increased accumulation of osmoprotectants and alteration of cellular fatty acid composition.

  7. Effect of multiple freeze-thaw cycles on the quality of instant sea cucumber: Emphatically on water status of by LF-NMR and MRI.

    Science.gov (United States)

    Tan, Mingqian; Lin, Zhuyi; Zu, Yinxue; Zhu, Beiwei; Cheng, Shasha

    2018-07-01

    Instant sea cucumber has become one popular product due to its convenience to eat, favourable taste and minimal loss of nutrients and bioactive components. However, there was rare information about the water dynamic of instant sea cucumber subjected to multiple freeze-thaw cycles. In this study, low-field nuclear magnetic resonance (LF-NMR) and magnetic resonance image (MRI) were employed to investigate the effect of freeze-thaw cycles on water status of instant sea cucumber. Four water populations corresponding to strongly bound water, weakly bound water, immobile water and free water were observed in instant sea cucumber. With the increase of freeze-thaw cycles, the transverse relaxation time of immobile and free water increased, while the peak area of free water decreased significantly. MRI enabled the visualization of water migration of instant sea cucumber during multiple freeze-thaw cycles. Multiple freeze-thaw cycles also led to significant changes of other quality properties including thawing loss, WHC, color parameters, texture and protein content, and enlarge the interspace between fiber network in microstructure. Good correlations between T 22 , A 22 , A 23 and thaw loss, WHC, L*, hardness and collagen content (0.873 ≤ r ≤ 0.958) revealed LF-NMR may be an effective real-time monitoring method of these physicochemical parameters as a non-destructive technique. Copyright © 2018 Elsevier Ltd. All rights reserved.

  8. Polymerization with freezing

    International Nuclear Information System (INIS)

    Ben-Naim, E; Krapivsky, P L

    2005-01-01

    Irreversible aggregation processes involving reactive and frozen clusters are investigated using the rate equation approach. In aggregation events, two clusters join irreversibly to form a larger cluster; additionally, reactive clusters may spontaneously freeze. Frozen clusters do not participate in merger events. Generally, freezing controls the nature of the aggregation process, as demonstrated by the final distribution of frozen clusters. The cluster mass distribution has a power-law tail, F k ∼k -γ , when the freezing process is sufficiently slow. Different exponents, γ = 1 and 3, are found for the constant and the product aggregation rates, respectively. For the latter case, the standard polymerization model, either no gels, or a single gel, or even multiple gels, may be produced

  9. Generalized structural theory of freezing

    International Nuclear Information System (INIS)

    Yussouff, M.

    1980-10-01

    The first-principles order parameter theory of freezing, proposed in an earlier work, has been successful in yielding quantitative agreement with known freezing parameters for monoatomic liquids forming solids with one atom per unit cell. A generalization of this theory is presented here to include the effects of a basis set of many atoms per unit cell. The basic equations get modified by the 'density structure factors' fsub(i) which arise from the density variations within the unit cell. Calculations are presented for the important case of monoatomic liquids freezing into hexagonal close packed solids. It is concluded that all freezing transitions can be described by using structural correlations in the liquid instead of the pair potential; and that the three body correlations are important in deciding the type of solid formed after freezing. (author)

  10. Anhydrobiosis and freezing-tolerance: adaptations that facilitate the establishment of Panagrolaimus nematodes in polar habitats.

    Science.gov (United States)

    McGill, Lorraine M; Shannon, Adam J; Pisani, Davide; Félix, Marie-Anne; Ramløv, Hans; Dix, Ilona; Wharton, David A; Burnell, Ann M

    2015-01-01

    Anhydrobiotic animals can survive the loss of both free and bound water from their cells. While in this state they are also resistant to freezing. This physiology adapts anhydrobiotes to harsh environments and it aids their dispersal. Panagrolaimus davidi, a bacterial feeding anhydrobiotic nematode isolated from Ross Island Antarctica, can survive intracellular ice formation when fully hydrated. A capacity to survive freezing while fully hydrated has also been observed in some other Antarctic nematodes. We experimentally determined the anhydrobiotic and freezing-tolerance phenotypes of 24 Panagrolaimus strains from tropical, temperate, continental and polar habitats and we analysed their phylogenetic relationships. We found that several other Panagrolaimus isolates can also survive freezing when fully hydrated and that tissue extracts from these freezing-tolerant nematodes can inhibit the growth of ice crystals. We show that P. davidi belongs to a clade of anhydrobiotic and freezing-tolerant panagrolaimids containing strains from temperate and continental regions and that P. superbus, an early colonizer at Surtsey island, Iceland after its volcanic formation, is closely related to a species from Pennsylvania, USA. Ancestral state reconstructions show that anhydrobiosis evolved deep in the phylogeny of Panagrolaimus. The early-diverging Panagrolaimus lineages are strongly anhydrobiotic but weakly freezing-tolerant, suggesting that freezing tolerance is most likely a derived trait. The common ancestors of the davidi and the superbus clades were anhydrobiotic and also possessed robust freezing tolerance, along with a capacity to inhibit the growth and recrystallization of ice crystals. Unlike other endemic Antarctic nematodes, the life history traits of P. davidi do not show evidence of an evolved response to polar conditions. Thus we suggest that the colonization of Antarctica by P. davidi and of Surtsey by P. superbus may be examples of recent "ecological fitting

  11. Anhydrobiosis and freezing-tolerance: adaptations that facilitate the establishment of Panagrolaimus nematodes in polar habitats.

    Directory of Open Access Journals (Sweden)

    Lorraine M McGill

    Full Text Available Anhydrobiotic animals can survive the loss of both free and bound water from their cells. While in this state they are also resistant to freezing. This physiology adapts anhydrobiotes to harsh environments and it aids their dispersal. Panagrolaimus davidi, a bacterial feeding anhydrobiotic nematode isolated from Ross Island Antarctica, can survive intracellular ice formation when fully hydrated. A capacity to survive freezing while fully hydrated has also been observed in some other Antarctic nematodes. We experimentally determined the anhydrobiotic and freezing-tolerance phenotypes of 24 Panagrolaimus strains from tropical, temperate, continental and polar habitats and we analysed their phylogenetic relationships. We found that several other Panagrolaimus isolates can also survive freezing when fully hydrated and that tissue extracts from these freezing-tolerant nematodes can inhibit the growth of ice crystals. We show that P. davidi belongs to a clade of anhydrobiotic and freezing-tolerant panagrolaimids containing strains from temperate and continental regions and that P. superbus, an early colonizer at Surtsey island, Iceland after its volcanic formation, is closely related to a species from Pennsylvania, USA. Ancestral state reconstructions show that anhydrobiosis evolved deep in the phylogeny of Panagrolaimus. The early-diverging Panagrolaimus lineages are strongly anhydrobiotic but weakly freezing-tolerant, suggesting that freezing tolerance is most likely a derived trait. The common ancestors of the davidi and the superbus clades were anhydrobiotic and also possessed robust freezing tolerance, along with a capacity to inhibit the growth and recrystallization of ice crystals. Unlike other endemic Antarctic nematodes, the life history traits of P. davidi do not show evidence of an evolved response to polar conditions. Thus we suggest that the colonization of Antarctica by P. davidi and of Surtsey by P. superbus may be examples of recent

  12. Comparison of real-time reverse transcriptase polymerase chain reaction of peripheral blood mononuclear cells, serum and cell-free body cavity effusion for the diagnosis of feline infectious peritonitis.

    Science.gov (United States)

    Doenges, Stephanie J; Weber, Karin; Dorsch, Roswitha; Fux, Robert; Hartmann, Katrin

    2017-04-01

    Objectives Diagnosis of feline infectious peritonitis (FIP) remains challenging, especially in cats without effusions. The objective of this study was to evaluate the sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction (RT-PCR) detecting feline coronavirus (FCoV) RNA in peripheral blood mononuclear cells (PBMCs) and serum in comparison with the same real-time RT-PCR in cell-free body cavity effusion. Methods This prospective case-control study included 92 cats. Forty-three cats had a definitive diagnosis of FIP, established either by histopathological examination (n = 28) or by positive immunofluorescence staining of FCoV antigen in macrophages of effusions (n = 11), or by both methods (n = 4). Forty-nine control cats had other diseases but similar clinical signs. Real-time RT-PCR was performed on PBMCs of 37 cats (21 cats with FIP, 16 controls), on serum of 51 cats (26 cats with FIP, 25 controls) and on cell-free body cavity effusion of 69 cats (36 cats with FIP, 33 controls). Sensitivity, specificity, positive and negative predictive value, including 95% confidence intervals (CI), were calculated. Results Real-time RT-PCR of PBMCs, serum and cell-free body cavity effusion showed a specificity of 100% (95% CI 79.4-100% in PBMCs, 86.3-100% in serum, 89.4-100% in cell-free body cavity effusion) and a sensitivity of 28.6% (95% CI 11.3-52.2%) in PBMCs, 15.4% (95% CI 4.4-34.9%) in serum and 88.9% (95% CI 73.9-96.9%) in cell-free body cavity effusion to diagnose FIP. Conclusions and relevance Although it is known that RT-PCR can often provide false-positive results in healthy cats, this real-time RT-PCR was shown to be a specific tool for the diagnosis of FIP when applied in a clinical setting. Sensitivity in cell-free body cavity effusion was high but low in PBMCs and serum. PBMC samples showed a higher sensitivity than serum samples, and are therefore a better choice if no effusion is present.

  13. Influence of common preanalytical variations on the metabolic profile of serum samples in biobanks

    Energy Technology Data Exchange (ETDEWEB)

    Fliniaux, Ophelie [University of Picardie Jules Verne, Laboratoire de Phytotechnologie EA 3900-BioPI (France); Gaillard, Gwenaelle [Biobanque de Picardie (France); Lion, Antoine [University of Picardie Jules Verne, Laboratoire de Phytotechnologie EA 3900-BioPI (France); Cailleu, Dominique [Batiment Serres-Transfert, rue de Mai/rue Dallery, Plateforme Analytique (France); Mesnard, Francois, E-mail: francois.mesnard@u-picardie.fr [University of Picardie Jules Verne, Laboratoire de Phytotechnologie EA 3900-BioPI (France); Betsou, Fotini [Integrated Biobank of Luxembourg (Luxembourg)

    2011-12-15

    A blood pre-centrifugation delay of 24 h at room temperature influenced the proton NMR spectroscopic profiles of human serum. A blood pre-centrifugation delay of 24 h at 4 Degree-Sign C did not influence the spectroscopic profile as compared with 4 h delays at either room temperature or 4 Degree-Sign C. Five or ten serum freeze-thaw cycles also influenced the proton NMR spectroscopic profiles. Certain common in vitro preanalytical variations occurring in biobanks may impact the metabolic profile of human serum.

  14. A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells.

    Science.gov (United States)

    Ahmadian Baghbaderani, Behnam; Tian, Xinghui; Scotty Cadet, Jean; Shah, Kevan; Walde, Amy; Tran, Huan; Kovarcik, Don Paul; Clarke, Diana; Fellner, Thomas

    2016-01-01

    Human pluripotent stem cells (hPSCs) present an unprecedented opportunity to advance human health by offering an alternative and renewable cell resource for cellular therapeutics and regenerative medicine. The present demand for high quality hPSCs for use in both research and clinical studies underscores the need to develop technologies that will simplify the cultivation process and control variability. Here we describe the development of a robust, defined and xeno-free hPSC medium that supports reliable propagation of hPSCs and generation of human induced pluripotent stem cells (hiPSCs) from multiple somatic cell types; long-term serial subculturing of hPSCs with every-other-day (EOD) medium replacement; and banking fully characterized hPSCs. The hPSCs cultured in this medium for over 40 passages are genetically stable, retain high expression levels of the pluripotency markers TRA-1-60, TRA-1-81, Oct-3/4 and SSEA-4, and readily differentiate into ectoderm, mesoderm and endoderm. Importantly, the medium plays an integral role in establishing a cGMP-compliant process for the manufacturing of hiPSCs that can be used for generation of clinically relevant cell types for cell replacement therapy applications.

  15. The cytoplasmic Cu,Zn superoxide dismutase of saccharomyces cerevisiae is required for resistance to freeze-thaw stress. Generation of free radicals during freezing and thawing

    DEFF Research Database (Denmark)

    Park, J I; Grant, C M; Davies, Michael Jonathan

    1998-01-01

    The involvement of oxidative stress in freeze-thaw injury to yeast cells was analyzed using mutants defective in a range of antioxidant functions, including Cu,Zn superoxide dismutase (encoded by SOD1), Mn superoxide dismutase (SOD2), catalase A, catalase T, glutathione reductase, gamma...

  16. Freeze out in heavy ion reactions

    International Nuclear Information System (INIS)

    Csernai, Laszlo P.; Lazar, Zs.I.; Grassi, F.; Hama, Y.

    1998-01-01

    In fluid dynamical models the freeze out of particles across a three dimensional space-time hypersurface is discussed. The calculation of final momentum distribution of emitted particles is described for freeze out surfaces, with both space-like and time-like normals, taking into account conservation laws across the freeze out discontinuity. Generally the conservation laws lead to a change of temperature, baryon density and flow velocity at freeze out. (author)

  17. A Robust, Cell-free Production System for On-Demand Protein Synthesis in Space

    Data.gov (United States)

    National Aeronautics and Space Administration — We will develop a new cell-free expression system that functions after rehydrating from a freeze-dried condition. Freeze-dried powder that can be stored or...

  18. Renal clearance of an ionic high-osmolar and a nonionic low-osmolar contrast medium

    International Nuclear Information System (INIS)

    Thomsen, H.S.; Vestergaard, A.; Nielsen, S.L.; Fogh-Andersen, N.; Golman, K.; Dorph, S.

    1991-01-01

    One hundred patients with normal serum creatinine concentration underwent intravenous urography with either an ionic high-osmolar (diatrizoate) or a nonionic low-osmolar (iopamidol) contrast medium after randomization. Before injection of the contrast medium, a blood sample was drawn for determinating serum creatinine concentration, and a urine sample for measurement of urine osmolality. Using x-ray fluorescence, the plasma concentration of iodine (contrast medium) was determined on blood samples drawn approximately 3 and 4 hours after injection of the contrast medium. The glomerular filtration rate was calculated by two different formulas: one requiring only a single sample and one requiring at least two samples (standard). There were poor correlations between the standard contrast medium clearance and the serum creatinine concentration, the estimated creatinine clearance (calculated from a nomogram), as well as the urine osmolality. The 3-hour and the 4-hour single-sample values correlated well with the two-sample values for both contrast media. In patients with normal serum creatinine, the glomerular filtration rate determined by measuring the contrast medium concentration in a single plasma sample obtained at 3 hours, is almost identical to the value determined from two samples. Consequently, two samples are unnecessary

  19. Quantification of circulating cell-free DNA in the serum of patients with obstructive sleep apnea-hypopnea syndrome.

    Science.gov (United States)

    Ye, Liang; Ma, Guan-Hua; Chen, Ling; Li, Min; Liu, Jia-Lin; Yang, Kun; Li, Qing-Yun; Li, Ning; Wan, Huan-Ying

    2010-12-01

    Serum cell-free DNA concentrations have been reported to increase in many acute diseases as well as in some chronic conditions such as cancer and autoimmune diseases. The aim of this study was to examine whether serum DNA concentrations were elevated in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). The effects of nasal continuous positive airway pressure (nCPAP) on serum DNA were also investigated. One hundred twenty-seven people diagnosed with OSAHS by polysomnography (PSG) were admitted into the OSAHS group, and 52 subjects without OSAHS were recruited for the control group. The OSAHS group was further divided into mild, moderate, and severe OSAHS subgroups based on their apnea-hypopnea index (AHI) during sleep. Ten patients with moderate and severe OSAHS were treated with nCPAP. Serum DNA, interleukin-6 (IL-6), and malonaldehyde (MDA) concentrations were measured and were found to be significantly higher in patients with moderate and severe OSAHS groups than those in the mild OSAHS and control groups (p DNA correlated positively with AHI, oxygen desaturation index (ODI), IL-6, and MDA, and negatively correlated with minimal oxygen saturation (miniSaO(2)) (all p DNA concentrations. After 6 months of nCPAP therapy, serum concentrations of DNA, IL-6, and MDA were significantly decreased (p DNA in patients with OSAHS was positively correlated with disease severity. Serum DNA may become an important parameter for monitoring the severity of OSAHS and effectiveness of therapy.

  20. Freeze-dried microarterial allografts

    International Nuclear Information System (INIS)

    Raman, J.; Hargrave, J.C.

    1990-01-01

    Rehydrated freeze-dried microarterial allografts were implanted to bridge arterial defects using New Zealand White rabbits as the experimental model. Segments of artery from the rabbit ear and thigh were harvested and preserved for a minimum of 2 weeks after freeze-drying. These allografts, approximately 1 mm in diameter and ranging from 1.5 to 2.5 cm in length, were rehydrated and then implanted in low-pressure and high-pressure arterial systems. Poor patency was noted in low-pressure systems in both allografts and autografts, tested in 12 rabbits. In the high-pressure arterial systems, allografts that were freeze-dried and reconstituted failed in a group of 10 rabbits with an 8-week patency rate of 30 percent. Gamma irradiation in an effort to reduce infection and antigenicity of grafts after freeze-drying was associated with a patency rate of 10 percent at 8 weeks in this system in another group of 10 rabbits. Postoperative cyclosporin A therapy was associated with a patency rate of 22.2 percent in the high-pressure arterial system in a 9-rabbit group. Control autografts in this system in a group of 10 rabbits showed a 100 percent patency at 8 weeks. Microarterial grafts depend on perfusion pressure of the vascular bed for long-term patency. Rehydrated freeze-dried microarterial allografts do not seem to function well in lengths of 1 to 2.5 cm when implanted in a high-pressure arterial system. Freeze-dried arterial allografts are probably not antigenic

  1. Parameterizing the competition between homogeneous and heterogeneous freezing in ice cloud formation – polydisperse ice nuclei

    Directory of Open Access Journals (Sweden)

    D. Barahona

    2009-08-01

    Full Text Available This study presents a comprehensive ice cloud formation parameterization that computes the ice crystal number, size distribution, and maximum supersaturation from precursor aerosol and ice nuclei. The parameterization provides an analytical solution of the cloud parcel model equations and accounts for the competition effects between homogeneous and heterogeneous freezing, and, between heterogeneous freezing in different modes. The diversity of heterogeneous nuclei is described through a nucleation spectrum function which is allowed to follow any form (i.e., derived from classical nucleation theory or from observations. The parameterization reproduces the predictions of a detailed numerical parcel model over a wide range of conditions, and several expressions for the nucleation spectrum. The average error in ice crystal number concentration was −2.0±8.5% for conditions of pure heterogeneous freezing, and, 4.7±21% when both homogeneous and heterogeneous freezing were active. The formulation presented is fast and free from requirements of numerical integration.

  2. Correlation of serum androgens and pituitary hormone levels with serum PSA less than 2.5 ng/ml.

    Science.gov (United States)

    Sofikerim, Mustafa; Oruç, Ozgür; Eskicorapci, Sadettin; Guliyev, Fuat; Ozen, Haluk

    2007-07-27

    The aim of this clinical study was to determine whether there is a relationship between total serum testosterone, free testosterone, FSH (Follicle-Stimulating Hormone), LH (Luteinizing Hormone) and serum prostate specific antigen (PSA) levels. We postulated that such a correlation existed then the use of hormone specific reference ranges might enhance the usefullness of PSA concentrations 40 years of age visiting our urology outpatient clinics. PSA was correlated to age (r = 0.23, p = 0.019), but there none between serum testosterone and age. No significant correlation was noted between testosterone or free testosterone and serum PSA levels, and none between serum FSH or LH and PSA. In age specific reference groups (41-49; 50-59; 60-69 years), we found no significant correlation between PSA and hormone concentrations. In this population of eugonadal men with serum PSA values less than 2.5 ng/ml, serum androgens and pituitary hormones do not appear to correlate with serum PSA.

  3. Application of the Quality by Design Approach to the Freezing Step of Freeze-Drying: Building the Design Space.

    Science.gov (United States)

    Arsiccio, Andrea; Pisano, Roberto

    2018-06-01

    The present work shows a rational method for the development of the freezing step of a freeze-drying cycle. The current approach to the selection of freezing conditions is still empirical and nonsystematic, thus resulting in poor robustness of control strategy. The final aim of this work is to fill this gap, describing a rational procedure, based on mathematical modeling, for properly choosing the freezing conditions. Mechanistic models are used for the prediction of temperature profiles during freezing and dimension of ice crystals being formed. Mathematical description of the drying phase of freeze-drying is also coupled with the results obtained by freezing models, thus providing a comprehensive characterization of the lyophilization process. In this framework, deep understanding of the phenomena involved is required, and according to the Quality by Design approach, this knowledge can be used to build the design space. The step-by-step procedure for building the design space for freezing is thus described, and examples of applications are provided. The calculated design space is validated upon experimental data, and we show that it allows easy control of the freezing process and fast selection of appropriate operating conditions. Copyright © 2018 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  4. Free-air ionization chamber, FAC-IR-300, designed for medium energy X-ray dosimetry

    International Nuclear Information System (INIS)

    Mohammadi, S.M.; Tavakoli-Anbaran, H.; Zeinali, H.Z.

    2017-01-01

    The primary standard for X-ray photons is based on parallel-plate free-air ionization chamber (FAC). Therefore, the Atomic Energy Organization of Iran (AEOI) is tried to design and build the free-air ionization chamber, FAC-IR-300, for low and medium energy X-ray dosimetry. The main aim of the present work is to investigate specification of the FAC-IR-300 ionization chamber and design it. FAC-IR-300 dosimeter is composed of two parallel plates, a high voltage (HV) plate and a collector plate, along with a guard electrode that surrounds the collector plate. The guard plate and the collector were separated by an air gap. For obtaining uniformity in the electric field distribution, a group of guard strips was used around the ionization chamber. These characterizations involve determining the exact dimensions of the ionization chamber by using Monte Carlo simulation and introducing correction factors.

  5. Cognitive Factors Affecting Freeze-like Behavior in Humans.

    Science.gov (United States)

    Alban, Michael W; Pocknell, Victoria

    2017-01-01

    Contemporary research on survival-related defensive behaviors has identified physiological markers of freeze/flight/fight. Our research focused on cognitive factors associated with freeze-like behavior in humans. Study 1 tested if an explicit decision to freeze is associated with the psychophysiological state of freezing. Heart rate deceleration occurred when participants chose to freeze. Study 2 varied the efficacy of freezing relative to other defense options and found "freeze" was responsive to variations in the perceived effectiveness of alternative actions. Study 3 tested if individual differences in motivational orientation affect preference for a "freeze" option when the efficacy of options is held constant. A trend in the predicted direction suggested that naturally occurring cognitions led loss-avoiders to select "freeze" more often than reward-seekers. In combination, our attention to the cognitive factors affecting freeze-like behavior in humans represents a preliminary step in addressing an important but neglected research area.

  6. Effects of Oxidized Glutathione, Bovine Serum Albumin, Cysteine and Lycopene on the Quality of Frozen-Thawed Ram Semen

    Directory of Open Access Journals (Sweden)

    O. Uysal

    2007-01-01

    Full Text Available Free radicals are known to be involved in lipid peroxidation as well as DNA and sperm membrane damages that may lead to decreased sperm motility or cell death. The balance between free radical production and their detoxification may be an important factor in sperm survival and function before, during and after cryopreservation. The aim of this study was to determine the effects of the addition of the antioxidants of oxidized glutathione (GSSG, bovine serum albumin (BSA, cysteine and lycopene to freezing media on the post-thawing sperm characteristics, including motility, morphology, acrosome integrity, viability and membrane integrity. A total number of 42 ejaculates were collected using the artifi cial vagina from 4 Akkaraman rams and 10 replicates of the ejaculates were diluted with a Tris-based extender containing additives and no additives as control. GSSG (5 mM, BSA (20 mg/ml, cysteine (10 mM and lycopene (800 μg showed more positive effects than other concentrations of the supplements and controls in protecting sperm characteristics after the freezing-thawing process (P < 0.001. Many aspects of sperm protection, e.g. sperm motility, viability and membrane stabilisation of the sperm cells during relative cryopreservation, are the key factors in determining the preservation of sperm function. The results of this study provide a new approach to the cryopreservation of sperm from rams and related breeds, and thereby contribute to the improvement of these breeds for the world sheep industry.

  7. Physical and chemical characteristics of meat from broilers raised in 4 different rearing systems, stored under freezing for up to 12 months.

    Science.gov (United States)

    Giampietro-Ganeco, A; Owens, C M; Mello, J L M; Souza, R A; Ferrari, F B; Souza, P A; Borba, H

    2017-10-01

    This study evaluated the effects of freezing (-18°C) for 12 mo on attributes related to the texture of breast, drumstick, and thigh from broilers raised in 4 different rearing systems. Five-hundred carcasses of male broilers raised in 4 rearing systems (Antibiotic-free, Cobb 500, n = 125; Free-range, Hubbard ISA, n = 125; Conventional, Cobb 500, n = 125; Organic, Cobb 500, n = 125) were divided into breast, drumstick, and thigh and stored under freezing (-18°C) for 3, 6, 9, and 12 months. Breast, drumstick, and thigh meat from broilers raised in all studied rearing systems showed reduction (P meat from alternative broilers showed an increase in shear force values, while breast meat from conventional broilers became tenderer during the freezing storage. In general, alternative broilers had harder thigh meat than conventional broilers. A reduction (P meat. Freezing chicken meat cuts for long periods, regardless of the rearing system, may interfere with the meat texture during preparation and consumption and, consequently, influence the consumer decision in a next purchase. © 2017 Poultry Science Association Inc.

  8. Cathepsin activities and membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling.

    Science.gov (United States)

    Zhang, T; Rawson, D M; Tosti, L; Carnevali, O

    2008-04-01

    This study investigated enzymatic activity of cathepsins and the membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling. Stage III oocytes (>0.5mm), obtained through dissection of anaesthetised female fish and desegregation of ovarian cumulus, were exposed to 2M methanol or 2M DMSO (both prepared in Hank's medium) for 30min at 22 degrees C before being loaded into 0.5ml plastic straws and placed into a programmable cooler. After controlled slow freezing, samples were plunged into liquid nitrogen (LN) and held for at least 10min, and thawed by immersing straws into a 27 degrees C water bath for 10s. Thawed oocytes were washed twice in Hank's medium. Cathepsin activity and membrane integrity of oocytes were assessed both after cryoprotectant treatment at 22 degrees C and after freezing in LN. Cathepsin B and L colorimetric analyses were performed using substrates Z-Arg-ArgNNap and Z-Phe-Arg-4MbetaNA-HCl, respectively, and 2-naphthylamine and 4-methoxy-2-naphthylamine were used as standards. Cathepsin D activity was performed by analysing the level of hydrolytic action on haemoglobin. Oocytes membrane integrity was assessed using 0.2% Trypan blue staining for 5min. Analysis of cathepsin activities showed that whilst the activity of cathepsin B and D was not affected by 2M DMSO treatment, their activity was lowered when treated with 2M methanol. Following freezing to -196 degrees C, the activity of all cathepsins (B, D and L) was significantly decreased in both 2M DMSO and 2M methanol. Trypan blue staining showed that 63.0+/-11.3% and 72.7+/-5.2% oocytes membrane stayed intact after DMSO and methanol treatment for 30min at 22 degrees C, respectively, whilst 14.9+/-2.6% and 1.4+/-0.8% stayed intact after freezing in DMSO and methanol to -196 degrees C. The results indicate that cryoprotectant treatment and freezing modified the activities of lysosomal enzymes involved in oocyte maturation and yolk

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

    Science.gov (United States)

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

    2015-03-13

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

  10. Local blood-brain barrier penetration following systemic contrast medium administration

    International Nuclear Information System (INIS)

    Utz, R.; Ekholm, S.E.; Isaac, L.; Sands, M.; Fonte, D.

    1988-01-01

    The present study was initiated by a severe complication in a patient with renal dysfunction who developed cortical blindness and weakness of her left extremities 30 hours following renal and abdominal angiography. To evaluate the impact of prolonged high serum concentrations of contrast medium (CM) this clinical situation was simulated in a laboratory model using sheep with elevated serum levels of contrast medium maintained for 48 hours. The experimental data did not support the theory that the prolonged exposure to high circulating levels of contrast medium (4 ml/kg body weight of meglumine diatrizoate 60%) is sufficient alone to cause penetration of the blood-brain barrier. (orig.)

  11. Freeze-drying of HI-6-loaded recombinant human serum albumin nanoparticles for improved storage stability.

    Science.gov (United States)

    Dadparvar, Miriam; Wagner, Sylvia; Wien, Sascha; Worek, Franz; von Briesen, Hagen; Kreuter, Jörg

    2014-10-01

    Severe intoxications with organophosphates require the immediate administration of atropine in combination with acetyl cholinesterase (AChE) reactivators such as HI-6. Although this therapy regimen enables the treatment of peripheral symptoms, the blood-brain barrier (BBB) restricts the access of the hydrophilic antidotes to the central nervous system which could lead to a fatal respiratory arrest. Therefore, HI-6-loaded albumin nanoparticles were previously developed to enhance the transport across this barrier and were able to reactivate organophosphate-(OP)-inhibited AChE in an in vitro BBB model. Since HI-6 is known to be moisture-sensitive, the feasibility of freeze-drying of the HI-6-loaded nanoparticles was investigated in the present study using different cryo- and lyoprotectants at different concentrations. Trehalose and sucrose (3%, w/v)-containing formulations were superior to mannitol concerning the physicochemical parameters of the nanoparticles whereas trehalose-containing samples were subject of a prolonged storage stability study at temperatures between -20°C and +40°C for predetermined time intervals. Shelf-life computations of the freeze-dried HI-6 nanoparticle formulations revealed a shelf-life time of 18 months when stored at -20°C. The formulations' efficacy was proven in vitro by reactivation of OP-inhibited AChE after transport over a porcine brain capillary endothelial cell layer model. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. 9 CFR 590.536 - Freezing operations.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Freezing operations. 590.536 Section 590.536 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG..., and Facility Requirements § 590.536 Freezing operations. (a) Freezing rooms shall be kept clean and...

  13. 9 CFR 590.534 - Freezing facilities.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Freezing facilities. 590.534 Section 590.534 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG..., and Facility Requirements § 590.534 Freezing facilities. (a) Freezing rooms, either on or off the...

  14. Response of Soil Biogeochemistry to Freeze-thaw Cycles: Impacts on Greenhouse Gas Emission and Nutrient Fluxes

    Science.gov (United States)

    Rezanezhad, F.; Parsons, C. T.; Smeaton, C. M.; Van Cappellen, P.

    2014-12-01

    Freeze-thaw is an abiotic stress applied to soils and is a natural process at medium to high latitudes. Freezing and thawing processes influence not only the physical properties of soil, but also the metabolic activity of soil microorganisms. Fungi and bacteria play a crucial role in soil organic matter degradation and the production of greenhouse gases (GHG) such as CO2, CH4 and N2O. Production and consumption of these atmospheric trace gases are the result of biological processes such as photosynthesis, aerobic respiration (CO2), methanogenesis, methanotrophy (CH4), nitrification and denitrification (N2O). To enhance our understanding of the effects of freeze-thaw cycles on soil biogeochemical transformations and fluxes, a highly instrumented soil column experiment was designed to realistically simulate freeze-thaw dynamics under controlled conditions. Pore waters collected periodically from different depths of the column and solid-phase analyses on core material obtained at the initial and end of the experiment highlighted striking geochemical cycling. CO2, CH4 and N2O production at different depths within the column were quantified from dissolved gas concentrations in pore water. Subsequent emissions from the soil surface were determined by direct measurement in the head space. Pulsed CO2 emission to the headspace was observed at the onset of thawing, however, the magnitude of the pulse decreased with each subsequent freeze-thaw cycle indicating depletion of a "freeze-thaw accessible" carbon pool. Pulsed CO2 emission was due to a combination of physical release of gases dissolved in porewater and entrapped below the frozen zone and changing microbial respiration in response to electron acceptor variability (O2, NO3-, SO42-). In this presentation, we focus on soil-specific physical, chemical, microbial factors (e.g. redox conditions, respiration, fermentation) and the mechanisms that drive GHG emission and nutrient cycling in soils under freeze-thaw cycles.

  15. 3 CFR - Pay Freeze

    Science.gov (United States)

    2010-01-01

    ... 3 The President 1 2010-01-01 2010-01-01 false Pay Freeze Presidential Documents Other Presidential Documents Memorandum of January 21, 2009 Pay Freeze Memorandum for the Assistant to the President and Chief... the White House staff forgo pay increases until further notice. Accordingly, as a signal of our shared...

  16. Free insulin-like growth factor I serum levels in 1430 healthy children and adults, and its diagnostic value in patients suspected of growth hormone deficiency

    DEFF Research Database (Denmark)

    Juul, A; Holm, K; Kastrup, K W

    1997-01-01

    fraction of the total IGF-I circulates in its free form, which is believed to be the biologically active form. However, our knowledge of the clinical or physiological value of determination of free IGF-I in serum is limited at present. In adults, the diagnostic value of total IGF-I and IGFBP-3......, commercially available immunoradiometric assay (Diagnostic Systems Laboratories) to establish valid normative data for this analysis. We studied the diagnostic value of free IGF-I in relation to total IGF-I and IGFBP-3 determinations in adults who were suspected of GHD. A GH provocative test, using oral...... determinations in patients suspected of GHD has only been reported in a few studies, whereas no previous reports on the diagnostic value of free IGF-I levels in adults suspected of GHD exist. Serum levels of free IGF-I were determined in 1430 healthy children, adolescents, and adults by a newly developed...

  17. Free insulin-like growth factor I serum levels in 1430 healthy children and adults, and its diagnostic value in patients suspected of growth hormone deficiency

    DEFF Research Database (Denmark)

    Juul, A; Holm, K; Kastrup, K W

    1997-01-01

    fraction of the total IGF-I circulates in its free form, which is believed to be the biologically active form. However, our knowledge of the clinical or physiological value of determination of free IGF-I in serum is limited at present. In adults, the diagnostic value of total IGF-I and IGFBP-3...... determinations in patients suspected of GHD has only been reported in a few studies, whereas no previous reports on the diagnostic value of free IGF-I levels in adults suspected of GHD exist. Serum levels of free IGF-I were determined in 1430 healthy children, adolescents, and adults by a newly developed......, commercially available immunoradiometric assay (Diagnostic Systems Laboratories) to establish valid normative data for this analysis. We studied the diagnostic value of free IGF-I in relation to total IGF-I and IGFBP-3 determinations in adults who were suspected of GHD. A GH provocative test, using oral...

  18. Effects of culture media conditions on production of eggs fertilized in vitro of embryos derived from ovary of high grade Hanwoo

    Directory of Open Access Journals (Sweden)

    Jun Young Lee

    2016-03-01

    Full Text Available Abstract Background This study was investigated the effects of culture media conditions on production of eggs fertilized in vitro of embryos from ovaries of high grade Korean native cow, Hanwoo. Methods The IVMD 101 and IVF 100 were used for in vitro maturation of selected Hanwoo oocytes and In vitro embryo culture after in vitro fertilization, respectively. The IVMD 101 and IVD 101 were used for in vitro culture and completely free of serum. Results The cleavage rates of 2-cell embryos in reference to Hanwoo oocytes were 86.7, 92.9 , and 90.1 % in the control group, IVDM101 medium and IVD101 medium, respectively which indicates that the IVDM101 medium and IVD101 medium may result favorable outcomes. The in vitro development rates of blastocysts were 12.4, 38.4 and 32.4 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. For hatched blastocysts, it was 5.3, 33.9, and 28.6 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. Hence, more favorable results were expected for the hatched blastocysts in which the IVMD101 medium and IVD101 medium were used than the control group. Average cell numbers of blastocysts were 128.3, 165.7, and 163.6 in the groups of TCM-199 + 10 % FBS medium, IVMD 101 medium, and IVD 101 medium, respectively which clearly show that the IVMD 101 and IVD 101 medium consequence significantly higher cell numbers compared to the control group (i.e., TCM-199 + 10 % FBS medium. Pregnancy rate after embryo transfer was 39.6 % when the serum free medium was used which is higher than that of the medium supplemented with serum (32.8 %. In addition, stillbirth rates were 4.9 % in the group of serum free medium whereas it was 13.6 % in the serum supplemented medium (13.6 %. Conclusions Taken altogether, serum free media, the IVMD 101 and IVD 101 represented more favorable results in the embryo development rate of embryos, cell numbers of blastocyst, and

  19. Organotypic hippocampal slice culture from the adult mouse brain: a versatile tool for translational neuropsychopharmacology.

    Science.gov (United States)

    Kim, Hyunjeong; Kim, Eosu; Park, Minsun; Lee, Eun; Namkoong, Kee

    2013-03-05

    One of the most significant barriers towards translational neuropsychiatry would be an unavailability of living brain tissues. Although organotypic brain tissue culture could be a useful alternative enabling observation of temporal changes induced by various drugs in living brain tissues, a proper method to establish a stable organotypic brain slice culture system using adult (rather than neonatal) hippocampus has been still elusive. In this study, we evaluated our simple method using the serum-free culture medium for successful adult organotypic hippocampal slice culture. Several tens of hippocampal slices from a single adult mouse (3-5 months old) were cultured in serum-free versus serum-containing conventional culture medium for 30 days and underwent various experiments to validate the effects of the existence of serum in the culture medium. Neither the excessive regression of neuronal viability nor metabolic deficiency was observed in the serum-free medium culture in contrast to the serum-containing medium culture. Despite such viability, newly generated immature neurons were scarcely detected in the serum-free culture, suggesting that the original neurons in the brain slice persist rather than being replaced by neurogenesis. Key structural features of in vivo neural tissue constituting astrocytes, neural processes, and pre- and post-synapses were also well preserved in the serum-free culture. In conclusion, using the serum-free culture medium, the adult hippocampal slice culture system will serve as a promising ex vivo tool for various fields of neuroscience, especially for studies on aging-related neuropsychiatric disorders or for high throughput screening of potential agents working against such disorders. Copyright © 2012 Elsevier Inc. All rights reserved.

  20. Fabrication of Li2TiO3 pebbles by a freeze drying process

    International Nuclear Information System (INIS)

    Lee, Sang-Jin; Park, Yi-Hyun; Yu, Min-Woo

    2013-01-01

    Li 2 TiO 3 pebbles were successfully fabricated by using a freeze drying process. The Li 2 TiO 3 slurry was prepared using a commercial powder of particle size 0.5–1.5 μm and the pebble pre-form was prepared by dropping the slurry into liquid nitrogen through a syringe needle. The droplets were rapidly frozen, changing their morphology to spherical pebbles. The frozen pebbles were dried at −10 °C in vacuum. To make crack-free pebbles, some glycerin was employed in the slurry, and long drying time and a low vacuum condition were applied in the freeze drying process. In the process, the solid content in the slurry influenced the spheroidicity of the pebble green body. The dried pebbles were sintered at 1200 °C in an air atmosphere. The sintered pebbles showed almost 40% shrinkage. The sintered pebbles revealed a porous microstructure with a uniform pore distribution and the sintered pebbles were crushed under an average load of 50 N in a compressive strength test. In the present study, a freeze drying process for fabrication of spherical Li 2 TiO 3 pebbles is introduced. The processing parameters, such as solid content in the slurry and the conditions of freeze drying and sintering, are also examined

  1. Serum-free culture alters the quantity and protein composition of neuroblastoma-derived extracellular vesicles

    Directory of Open Access Journals (Sweden)

    Jinghuan Li

    2015-05-01

    Full Text Available Extracellular vesicles (EVs play a significant role in cell–cell communication in numerous physiological processes and pathological conditions, and offer promise as novel biomarkers and therapeutic agents for genetic diseases. Many recent studies have described different molecular mechanisms that contribute to EV biogenesis and release from cells. However, little is known about how external stimuli such as cell culture conditions can affect the quantity and content of EVs. While N2a neuroblastoma cells cultured in serum-free (OptiMEM conditions did not result in EVs with significant biophysical or size differences compared with cells cultured in serum-containing (pre-spun conditions, the quantity of isolated EVs was greatly increased. Moreover, the expression levels of certain vesicular proteins (e.g. small GTPases, G-protein complexes, mRNA processing proteins and splicing factors, some of which were previously reported to be involved in EV biogenesis, were found to be differentially expressed in EVs under different culture conditions. These data, therefore, contribute to the understanding of how extracellular factors and intracellular molecular pathways affect the composition and release of EVs.

  2. Diagnostic value of serum free PSA and the ratio of free to total PSA in the diagnosis of prostate cancer

    International Nuclear Information System (INIS)

    Qiu Ningyan; Zhang Jingxin; Wu Jinchang; Gong Yiming; Li Huiping

    2005-01-01

    In order to evaluate the value of free prostate specific antigen (FPSA) and F/T PSA ratio in differential diagnosis of benign prostate hyperplasia (BPH) from prostate cancer (PC), serum FPSA and TPSA levels were measured in 85 patients with PC, 97 BPH and 89 healthy volunteers by chemiluminescence enzyme immunoassay (CLEIA), and the ratio of F/T PSA was calculated. The results showed that serum FPSA and TPSA levels were increased in healthy volunteers of 41-88 years old and were significantly higher in healthy volunteers of 61-88 years old than that in 20-40 gear old (P 10.0 μg/L were 65.0%, 30.9% and 4.1%, respectively, while they were 5.9%, 20.0% and 74.1% in PC patients (P<0.01). When the TPSA value was between 4.0-10.0 μg/L and the ratio of F/T PSA was at 0.10 and below, the probability of PC was larger(88.9%). But the ratio of F/T PSA was at 0.25 and above, the probability of PC was smaller(6.20%). Serum FPSA and TPSA both increased with age in healthy volunteers of 41-88 years old and were positively correlated with age. There were about 30.9% of BPH and 20.0% of PC patients with overlapping of TPSA level. Our conclusion is that the F/T PSA ratio can significantly enhance the specificity for PC diagnosis, especially when the TPSA is within the diagnostic gray zone. (authors)

  3. Serum Free Light Chains

    Science.gov (United States)

    ... lab's website in order to provide you with background information about the test(s) you had performed. You will need to return ... the free light chains have a much shorter half-life (3-5 hours) than ... cell disorders, the test is also used for assessing response and minimal ...

  4. Isolation and characterization of Wharton’s jelly-derived multipotent mesenchymal stromal cells obtained from bovine umbilical cord and maintained in a defined serum-free three-dimensional system

    Directory of Open Access Journals (Sweden)

    Cardoso Tereza C

    2012-05-01

    Full Text Available Abstract Background The possibility for isolating bovine mesenchymal multipotent cells (MSCs from fetal adnexa is an interesting prospect because of the potential for these cells to be used for biotechnological applications. Bone marrow and adipose tissue are the most common sources of MSCs derived from adult animals. However, little knowledge exists about the characteristics of these progenitors cells in the bovine species. Traditionally most cell cultures are developed in two dimensional (2D environments. In mammalian tissue, cells connect not only to each other, but also support structures called the extracellular matrix (ECM. The three-dimensional (3D cultures may play a potential role in cell biotechnology, especially in tissue therapy. In this study, bovine-derived umbilical cord Wharton’s jelly (UC-WJ cells were isolated, characterized and maintained under 3D-free serum condition as an alternative of stem cell source for future cell banking. Results Bovine-derived UC-WJ cells, collected individually from 5 different umbilical cords sources, were successfully cultured under serum-free conditions and were capable to support 60 consecutive passages using commercial Stemline® mesenchymal stem cells expansion medium. Moreover, the UC-WJ cells were differentiated into osteocytes, chondrocytes, adipocytes and neural-like cells and cultured separately. Additionally, the genes that are considered important embryonic, POU5F1 and ITSN1, and mesenchymal cell markers, CD105+, CD29+, CD73+ and CD90+ in MSCs were also expressed in five bovine-derived UC-WJ cultures. Morphology of proliferating cells typically appeared fibroblast-like spindle shape presenting the same viability and number. These characteristics were not affected during passages. There were 60 chromosomes at the metaphase, with acrocentric morphology and intense telomerase activity. Moreover, the proliferative capacity of T cells in response to a mitogen stimulus was suppressed when

  5. Compound serum and hemin free medium for cultivation of ...

    African Journals Online (AJOL)

    Aghomotsegin

    2015-06-23

    Jun 23, 2015 ... parasite of the gecko Tarentolae annularis (Wallbanks et al., 1985) and has been introduced as a novel eukaryotic expression system for the production of recombinant proteins with mammalian-like post translational modifica- tion pattern (Fritsche et al., 2007). The protozoan is non-pathogenic to humans ( ...

  6. Compound serum and hemin free medium for cultivation of ...

    African Journals Online (AJOL)

    Aghomotsegin

    2015-06-23

    Jun 23, 2015 ... mammalian host within a vacuole with lysosomal features known as parasitophorous vacuole (Galvao-Quintao et al., 1990; Sacks, 1989). Leishmania tarentolae is a parasite of the gecko Tarentolae annularis (Wallbanks et al., 1985) and has been introduced as a novel eukaryotic expression system for the ...

  7. Stepwise Differentiation of Pluripotent Stem Cells into Osteoblasts Using Four Small Molecules under Serum-free and Feeder-free Conditions

    Directory of Open Access Journals (Sweden)

    Kosuke Kanke

    2014-06-01

    Full Text Available Pluripotent stem cells are a promising tool for mechanistic studies of tissue development, drug screening, and cell-based therapies. Here, we report an effective and mass-producing strategy for the stepwise differentiation of mouse embryonic stem cells (mESCs and mouse and human induced pluripotent stem cells (miPSCs and hiPSCs, respectively into osteoblasts using four small molecules (CHIR99021 [CHIR], cyclopamine [Cyc], smoothened agonist [SAG], and a helioxanthin-derivative 4-(4-methoxyphenylpyrido[4′,3′:4,5]thieno[2,3-b]pyridine-2-carboxamide [TH] under serum-free and feeder-free conditions. The strategy, which consists of mesoderm induction, osteoblast induction, and osteoblast maturation phases, significantly induced expressions of osteoblast-related genes and proteins in mESCs, miPSCs, and hiPSCs. In addition, when mESCs defective in runt-related transcription factor 2 (Runx2, a master regulator of osteogenesis, were cultured by the strategy, they molecularly recapitulated osteoblast phenotypes of Runx2 null mice. The present strategy will be a platform for biological and pathological studies of osteoblast development, screening of bone-augmentation drugs, and skeletal regeneration.

  8. Quantitative characterization of nanoparticle agglomeration within biological media

    International Nuclear Information System (INIS)

    Hondow, Nicole; Brydson, Rik; Wang, Peiyi; Holton, Mark D.; Brown, M. Rowan; Rees, Paul; Summers, Huw D.; Brown, Andy

    2012-01-01

    Quantitative analysis of nanoparticle dispersion state within biological media is essential to understanding cellular uptake and the roles of diffusion, sedimentation, and endocytosis in determining nanoparticle dose. The dispersion of polymer-coated CdTe/ZnS quantum dots in water and cell growth medium with and without fetal bovine serum was analyzed by transmission electron microscopy (TEM) and dynamic light scattering (DLS) techniques. Characterization by TEM of samples prepared by plunge freezing the blotted solutions into liquid ethane was sensitive to the dispersion state of the quantum dots and enabled measurement of agglomerate size distributions even in the presence of serum proteins where DLS failed. In addition, TEM showed a reduced packing fraction of quantum dots per agglomerate when dispersed in biological media and serum compared to just water, highlighting the effect of interactions between the media, serum proteins, and the quantum dots. The identification of a heterogeneous distribution of quantum dots and quantum dot agglomerates in cell growth medium and serum by TEM will enable correlation with the previously reported optical metrology of in vitro cellular uptake of this quantum dot dispersion. In this paper, we present a comparative study of TEM and DLS and show that plunge-freeze TEM provides a robust assessment of nanoparticle agglomeration state.

  9. Canalization of freeze tolerance in an alpine grasshopper.

    Science.gov (United States)

    Hawes, Timothy C

    2015-10-01

    In the Rock and Pillar Range, New Zealand, the alpine grasshopper, Sigaus australis Hutton, survives equilibrium freezing (EF) all-year round. A comparison of freeze tolerance (FT) in grasshoppers over four austral seasons for a 1 year period finds that: (a) the majority (>70%) of the sample population of grasshoppers survive single freeze-stress throughout the year; (b) exposure to increased freeze stress (multiple freeze-stress events) does not lead to a loss of freeze tolerance; and (c) responses to increased freeze stress reveal seasonal tuning of the FT adaptation to environmental temperatures. The Rock and Pillar sample population provides a clear example of the canalization of the FT adaptation. Seasonal variability in the extent of tolerance of multiple freezing events indicates that physiology is modulated to environmental temperatures by phenotypic plasticity - i.e. the FT adaptation is permanent and adjustable. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Pre-analytical sample quality: metabolite ratios as an intrinsic marker for prolonged room temperature exposure of serum samples.

    Directory of Open Access Journals (Sweden)

    Gabriele Anton

    Full Text Available Advances in the "omics" field bring about the need for a high number of good quality samples. Many omics studies take advantage of biobanked samples to meet this need. Most of the laboratory errors occur in the pre-analytical phase. Therefore evidence-based standard operating procedures for the pre-analytical phase as well as markers to distinguish between 'good' and 'bad' quality samples taking into account the desired downstream analysis are urgently needed. We studied concentration changes of metabolites in serum samples due to pre-storage handling conditions as well as due to repeated freeze-thaw cycles. We collected fasting serum samples and subjected aliquots to up to four freeze-thaw cycles and to pre-storage handling delays of 12, 24 and 36 hours at room temperature (RT and on wet and dry ice. For each treated aliquot, we quantified 127 metabolites through a targeted metabolomics approach. We found a clear signature of degradation in samples kept at RT. Storage on wet ice led to less pronounced concentration changes. 24 metabolites showed significant concentration changes at RT. In 22 of these, changes were already visible after only 12 hours of storage delay. Especially pronounced were increases in lysophosphatidylcholines and decreases in phosphatidylcholines. We showed that the ratio between the concentrations of these molecule classes could serve as a measure to distinguish between 'good' and 'bad' quality samples in our study. In contrast, we found quite stable metabolite concentrations during up to four freeze-thaw cycles. We concluded that pre-analytical RT handling of serum samples should be strictly avoided and serum samples should always be handled on wet ice or in cooling devices after centrifugation. Moreover, serum samples should be frozen at or below -80°C as soon as possible after centrifugation.

  11. Effects of a honeybee sting on the serum free amino acid profile in humans.

    Directory of Open Access Journals (Sweden)

    Jan Matysiak

    Full Text Available The aim of this study was to assess the response to a honeybee venom by analyzing serum levels of 34 free amino acids. Another goal of this study was to apply complex analytic-bioinformatic-clinical strategy based on up-to-date achievements of mass spectrometry in metabolomic profiling. The amino acid profiles were determined using hybrid triple quadrupole/linear ion trap mass spectrometer coupled with a liquid chromatography instrument. Serum samples were collected from 27 beekeepers within 3 hours after they were stung and after a minimum of 6 weeks following the last sting. The differences in amino acid profiles were evaluated using MetaboAnalyst and ROCCET web portals. Chemometric tests showed statistically significant differences in the levels of L-glutamine (Gln, L-glutamic acid (Glu, L-methionine (Met and 3-methyl-L-histidine (3MHis between the two analyzed groups of serum samples. Gln and Glu appeared to be the most important metabolites for distinguishing the beekeepers tested shortly after a bee sting from those tested at least 6 weeks later. The role of some amino acids in the response of an organism to the honeybee sting was also discussed. This study indicated that proposed methodology may allow to identify the individuals just after the sting and those who were stung at least 6 weeks earlier. The results we obtained will contribute to better understanding of the human body response to the honeybee sting.

  12. Influence of cell culture medium composition on in vitro dissolution behavior of a fluoride-containing bioactive glass.

    Science.gov (United States)

    Shah, Furqan A; Brauer, Delia S; Wilson, Rory M; Hill, Robert G; Hing, Karin A

    2014-03-01

    Bioactive glasses are used clinically for bone regeneration, and their bioactivity and cell compatibility are often characterized in vitro, using physiologically relevant test solutions. The aim of this study was to show the influence of varying medium characteristics (pH, composition, presence of proteins) on glass dissolution and apatite formation. The dissolution behavior of a fluoride-containing bioactive glass (BG) was investigated over a period of one week in Eagle's Minimal Essential Medium with Earle's Salts (MEM), supplemented with either, (a) acetate buffer, (b) 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, (c) HEPES + carbonate, or (d) HEPES + carbonate + fetal bovine serum. Results show pronounced differences in pH, ion release, and apatite formation over 1 week: Despite its acidic pH (pH 5.8 after BG immersion, as compared to pH 7.4-8.3 for HEPES-containing media), apatite formation was fastest in acetate buffered (HEPES-free) MEM. Presence of carbonate resulted in formation of calcite (calcium carbonate). Presence of serum proteins, on the other hand, delayed apatite formation significantly. These results confirm that the composition and properties of a tissue culture medium are important factors during in vitro experiments and need to be taken into consideration when interpreting results from dissolution or cell culture studies. Copyright © 2013 Wiley Periodicals, Inc.

  13. Gamma radiation effects on liofilized human serum

    International Nuclear Information System (INIS)

    Padron Soler, E.; Romay Penabad, Z.; Chavez Ardanza, A.; Hernandez Gonzalez, C.; Martin Gonzalez, O.; Garcia Gonzalez, I.; Prieto Miranda, E.

    1995-01-01

    Human freeze dried serum was artificially contaminated with Flavobacterium sp. for studying the effects of gamma radiation of it. The radiobiological parameters of the contaminator were determined and the sterilization dose was set. The quality of the product irradiated at both, calculated sterilization dose (8.5 kGy) an another one about 25 kGy was determined. It was made according to: sterility testing, total proteins, pH enzymes (alanina-aminotransferase, aspartato-aminotransferase, alkaline phosphatase), protein electrophoresis, fast performance liquid chromatographic and effect on the cellular growth. From the latter was concluded that the calculated sterilization dose was adequate form keeping the biological properties and viability of the irradiated serum. Nevertheless, the dose of 25 k Gy was not adequate because of its dangerous effects on the cell culture

  14. Effect of magnetic field on food freezing

    OpenAIRE

    村田, 圭治; 奥村, 太一; 荒賀, 浩一; 小堀, 康功

    2010-01-01

    [Abstract] This paper presents an experimental investigation on effects of magnetic field on food freezing process. Although purpose of food freezing is to suppress the deterioration of food, freezing breaks food tissue down, and some nutrient and delicious element flow out after thawing. Recently, a few of refrigeration equipments with electric and magnetic fields have attracted attention from food production companies and mass media. Water and tuna were freezed in magnetic field (100kH, 1.3...

  15. Investigation of the solubility and the potentials for purification of serum amyloid A (SAA) from equine acute phase serum

    DEFF Research Database (Denmark)

    Christensen, Michelle Brønniche; Sørensen, Jens Christian; Jacobsen, Stine

    2013-01-01

    BACKGROUND: Serum amyloid A (SAA) is useful as a diagnostic marker of systemic inflammation in horses, but only heterologous assays based on non-equine calibration and standardization are available for measurements of equine SAA. More accurate measurements could be obtained using purified species......-specific SAA in native conformation for assay calibration and standardization. Further knowledge about the biochemical properties of SAA would facilitate a future production of native species-specific calibration material Therefore, the aim of the study was an investigation of the solubility and potentials...... for purification of equine SAA based on biochemical properties.Freeze dried equine acute phase serum was dissolved in 70% 2-propanol, 8 M urea, and milli-Q water, respectively. Supercritical fluid extraction (SFE), size-exclusive chromatography (FPLC-SEC), and preparative isoelectric focusing (IEF) were performed...

  16. Freezing of Lennard-Jones-type fluids

    International Nuclear Information System (INIS)

    Khrapak, Sergey A.; Chaudhuri, Manis; Morfill, Gregor E.

    2011-01-01

    We put forward an approximate method to locate the fluid-solid (freezing) phase transition in systems of classical particles interacting via a wide range of Lennard-Jones-type potentials. This method is based on the constancy of the properly normalized second derivative of the interaction potential (freezing indicator) along the freezing curve. As demonstrated recently it yields remarkably good agreement with previous numerical simulation studies of the conventional 12-6 Lennard-Jones (LJ) fluid [S.A.Khrapak, M.Chaudhuri, G.E.Morfill, Phys. Rev. B 134, 052101 (2010)]. In this paper, we test this approach using a wide range of the LJ-type potentials, including LJ n-6 and exp-6 models, and find that it remains sufficiently accurate and reliable in reproducing the corresponding freezing curves, down to the triple-point temperatures. One of the possible application of the method--estimation of the freezing conditions in complex (dusty) plasmas with ''tunable'' interactions--is briefly discussed.

  17. Freeze-thaw durability of air-entrained concrete.

    Science.gov (United States)

    Shang, Huai-Shuai; Yi, Ting-Hua

    2013-01-01

    One of the most damaging actions affecting concrete is the abrupt temperature change (freeze-thaw cycles). The types of deterioration of concrete structures by cyclic freeze-thaw can be largely classified into surface scaling (characterized by the weight loss) and internal crack growth (characterized by the loss of dynamic modulus of elasticity). The present study explored the durability of concrete made with air-entraining agent subjected to 0, 100, 200, 300, and 400 cycles of freeze-thaw. The experimental study of C20, C25, C30, C40, and C50 air-entrained concrete specimens was completed according to "the test method of long-term and durability on ordinary concrete" GB/T 50082-2009. The dynamic modulus of elasticity and weight loss of specimens were measured after different cycles of freeze-thaw. The influence of freeze-thaw cycles on the relative dynamic modulus of elasticity and weight loss was analyzed. The findings showed that the dynamic modulus of elasticity and weight decreased as the freeze-thaw cycles were repeated. They revealed that the C30, C40, and C50 air-entrained concrete was still durable after 300 cycles of freeze-thaw according to the experimental results.

  18. Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum.

    Science.gov (United States)

    Ziegler, Anja; Everett, Helen; Hamza, Eman; Garbani, Mattia; Gerber, Vinzenz; Marti, Eliane; Steinbach, Falko

    2016-11-15

    Dendritic cells are professional antigen-presenting cells that play an essential role in the initiation and modulation of T cell responses. They have been studied widely for their potential clinical applications, but for clinical use to be successful, alternatives to xenogeneic substances like fetal bovine serum (FBS) in cell culture need to be found. Protocols for the generation of dendritic cells ex vivo from monocytes are well established for several species, including horses. Currently, the gold standard protocol for generating dendritic cells from monocytes across various species relies upon a combination of GM-CSF and IL-4 added to cell culture medium which is supplemented with FBS. The aim of this study was to substitute FBS with heterologous horse serum. For this purpose, equine monocyte-derived dendritic cells (eqMoDC) were generated in the presence of horse serum or FBS and analysed for the effect on morphology, phenotype and immunological properties. Changes in the expression of phenotypic markers (CD14, CD86, CD206) were assessed during dendritic cell maturation by flow cytometry. To obtain a more complete picture of the eqMoDC differentiation and assess possible differences between FBS- and horse serum-driven cultures, a transcriptomic microarray analysis was performed. Lastly, immature eqMoDC were primed with a primary antigen (ovalbumin) or a recall antigen (tetanus toxoid) and, after maturation, were co-cultured with freshly isolated autologous CD5 + T lymphocytes to assess their T cell stimulatory capacity. The microarray analysis demonstrated that eqMoDC generated with horse serum were indistinguishable from those generated with FBS. However, eqMoDC incubated with horse serum-supplemented medium exhibited a more characteristic dendritic cell morphology during differentiation from monocytes. A significant increase in cell viability was also observed in eqMoDC cultured with horse serum. Furthermore, eqMoDC generated in the presence of horse serum

  19. A Robust, Enzyme-Free Glucose Sensor Based on Lysine-Assisted CuO Nanostructures

    Directory of Open Access Journals (Sweden)

    Qurrat-ul-Ain Baloach

    2016-11-01

    Full Text Available The production of a nanomaterial with enhanced and desirable electrocatalytic properties is of prime importance, and the commercialization of devices containing these materials is a challenging task. In this study, unique cupric oxide (CuO nanostructures were synthesized using lysine as a soft template for the evolution of morphology via a rapid and boiled hydrothermal method. The morphology and structure of the synthesized CuO nanomaterial were characterized using scanning electron microscopy (SEM and X-ray diffraction (XRD, respectively. The prepared CuO nanostructures showed high potential for use in the electrocatalytic oxidation of glucose in an alkaline medium. The proposed enzyme-free glucose sensor demonstrated a robust response to glucose with a wide linear range and high sensitivity, selectivity, stability, and reproducibility. To explore its practical feasibility, the glucose content of serum samples was successfully determined using the enzyme-free sensor. An analytical recovery method was used to measure the actual glucose from the serum samples, and the results were satisfactory. Moreover, the presented glucose sensor has high chemical stability and can be reused for repetitive measurements. This study introduces an enzyme-free glucose sensor as an alternative tool for clinical glucose quantification.

  20. A defined and xeno-free culture method enabling the establishment of clinical-grade human embryonic, induced pluripotent and adipose stem cells.

    Directory of Open Access Journals (Sweden)

    Kristiina Rajala

    2010-04-01

    Full Text Available The growth of stem cells in in vitro conditions requires optimal balance between signals mediating cell survival, proliferation, and self-renewal. For clinical application of stem cells, the use of completely defined conditions and elimination of all animal-derived materials from the establishment, culture, and differentiation processes is desirable.Here, we report the development of a fully defined xeno-free medium (RegES, capable of supporting the expansion of human embryonic stem cells (hESC, induced pluripotent stem cells (iPSC and adipose stem cells (ASC. We describe the use of the xeno-free medium in the derivation and long-term (>80 passages culture of three pluripotent karyotypically normal hESC lines: Regea 06/015, Regea 07/046, and Regea 08/013. Cardiomyocytes and neural cells differentiated from these cells exhibit features characteristic to these cell types. The same formulation of the xeno-free medium is capable of supporting the undifferentiated growth of iPSCs on human feeder cells. The characteristics of the pluripotent hESC and iPSC lines are comparable to lines derived and cultured in standard undefined culture conditions. In the culture of ASCs, the xeno-free medium provided significantly higher proliferation rates than ASCs cultured in medium containing allogeneic human serum (HS, while maintaining the differentiation potential and characteristic surface marker expression profile of ASCs, although significant differences in the surface marker expression of ASCs cultured in HS and RegES media were revealed.Our results demonstrate that human ESCs, iPSCs and ASCs can be maintained in the same defined xeno-free medium formulation for a prolonged period of time while maintaining their characteristics, demonstrating the applicability of the simplified xeno-free medium formulation for the production of clinical-grade stem cells. The basic xeno-free formulation described herein has the potential to be further optimized for specific

  1. Bulk water freezing dynamics on superhydrophobic surfaces

    Science.gov (United States)

    Chavan, S.; Carpenter, J.; Nallapaneni, M.; Chen, J. Y.; Miljkovic, N.

    2017-01-01

    In this study, we elucidate the mechanisms governing the heat-transfer mediated, non-thermodynamic limited, freezing delay on non-wetting surfaces for a variety of characteristic length scales, Lc (volume/surface area, 3 mm commercial superhydrophobic spray coatings, showing a monotonic increase in freezing time with coating thickness. The added thermal resistance of thicker coatings was much larger than that of the nanoscale superhydrophobic features, which reduced the droplet heat transfer and increased the total freezing time. Transient finite element method heat transfer simulations of the water slab freezing process were performed to calculate the overall heat transfer coefficient at the substrate-water/ice interface during freezing, and shown to be in the range of 1-2.5 kW/m2K for these experiments. The results shown here suggest that in order to exploit the heat-transfer mediated freezing delay, thicker superhydrophobic coatings must be deposited on the surface, where the coating resistance is comparable to the bulk water/ice conduction resistance.

  2. Determination of mercury in human serum and packed blood cells by neutron activation analysis

    International Nuclear Information System (INIS)

    Versieck, J.; Vanballenberghe, L.; Wittoek, A.; Vermeir, G.; Vandecasteele, C.

    1990-01-01

    A method is described for the determination of mercury in human blood serum and packed blood cells employing neutron activation analysis. Great attention was devoted to the collection and manipulation of the samples. The accuracy and precision of the method were tested by analyzing biological reference materials and by comparing the concentrations measured in a number of serum samples to those obtained by another, independent technique (cold vapor atomic absorption spectrometry) in the same samples. The article reports the levels measured in blood serum and packed blood cells samples from 15 adult volunteers, as well as the figures determined in a open-quotes second-generationclose quotes biological reference material (freeze-dried human serum), prepared and conditioned at the University of Ghent

  3. Applicable technical method for freeze-substitution of high pressure ...

    African Journals Online (AJOL)

    bmshsj

    2011-11-02

    Quintana, 1994) are available for the microscopic visualization of intracellular organelles. Cryo- fixation, plunge freezing, propane jet freezing, cold metal block freezing, and high pressure freezing provide advantages over chemical ...

  4. Mechanisms of deterioration of nutrients. [of freeze dried foods

    Science.gov (United States)

    Karel, M.; Flink, J. M.

    1976-01-01

    Methods which produce freeze dried foods of improved quality were examined with emphasis on storage stability. Specific topics discussed include: microstructure of freeze dried systems, investigation of structural changes in freeze dried systems, artificial food matrices, osmotic preconcentration to yield improved quality freeze dried fruits, and storage stability of osmotically preconcentrated freeze dried fruits.

  5. Antimicrobial medium- and long-chain free fatty acids prevent PrfA-dependent activation of virulence genes in Listeria monocytogenes.

    Science.gov (United States)

    Sternkopf Lillebæk, Eva Maria; Lambert Nielsen, Stine; Scheel Thomasen, Rikke; Færgeman, Nils J; Kallipolitis, Birgitte H

    The foodborne pathogen Listeria monocytogenes is the causative agent of the invasive disease listeriosis. Infection by L. monocytogenes involves bacterial crossing of the intestinal barrier and intracellular replication in a variety of host cells. The PrfA protein is the master regulator of virulence factors required for bacterial entry, intracellular replication and cell-to-cell spread. PrfA-dependent activation of virulence genes occurs primarily in the blood and during intracellular infection. In contrast, PrfA does not play a significant role in regulation of virulence gene expression in the intestinal environment. In the gastrointestinal phase of infection, the bacterium encounters a variety of antimicrobial agents, including medium- and long-chain free fatty acids that are commonly found in our diet and as active components of bile. Here we show that subinhibitory concentrations of specific antimicrobial free fatty acids act to downregulate transcription of PrfA-activated virulence genes. Interestingly, the inhibitory effect is also evident in cells encoding a constitutively active variant of PrfA. Collectively, our data suggest that antimicrobial medium- and long-chain free fatty acids may act as signals to prevent PrfA-mediated activation of virulence genes in environments where PrfA activation is not required, such as in food and the gastrointestinal tract. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  6. Effect of serum proteins on polystyrene nanoparticle uptake and intracellular trafficking in endothelial cells

    International Nuclear Information System (INIS)

    Guarnieri, Daniela; Guaccio, Angela; Fusco, Sabato; Netti, Paolo A.

    2011-01-01

    The physico-chemical properties of nanoparticles (NPs), such as small dimensions, surface charge and surface functionalization, control their capability to interact with cells and, in particular, with sub-cellular components. This interaction can be also influenced by the adsorption of molecules present in biological fluids, like blood, on NP surface. Here, we analysed the effect of serum proteins on 49 and 100 nm red fluorescent polystyrene NP uptake in porcine aortic endothelial (PAE) cells, as a model for vascular transport. To this aim, NP uptake kinetic, endocytic pathway and intracellular trafficking were studied by monitoring NPs inside cells through confocal microscopy and multiple particle tracking (MPT). We demonstrated that NPs are rapidly internalized by cells in serum-free (SF) medium, according to a saturation kinetic. Conversely, in 10% foetal bovine serum-enriched (SE) medium, NP uptake rate results drastically reduced. Moreover, NP internalization depends on an active endocytic mechanism that does not involve clathrin- and caveolae-mediated vesicular transport, in both SE and SF media. Furthermore, MPT data indicate that NP intracellular trafficking is unaffected by protein presence. Indeed, approximately 50–60% of internalized NPs is characterized by a sub-diffusive behaviour, whereas the remaining fraction shows an active motion. These findings demonstrate that the unspecific protein adsorption on NP surface can affect cellular uptake in terms of internalization kinetics, but it is not effective in controlling active and cellular-mediated uptake mechanisms of NPs and their intracellular routes.

  7. Non-Darcy Free Convection of Power-Law Fluids Over a Two-Dimensional Body Embedded in a Porous Medium

    KAUST Repository

    El-Amin, Mohamed

    2010-11-27

    A boundary layer analysis was presented to study the non-Darcy-free convection of a power-law fluid over a non-isothermal two-dimensional body embedded in a porous medium. The Ostwald-de Waele power-law model was used to characterize the non-Newtonian fluid behavior. Similarity solutions were obtained with variations in surface temperature or surface heat flux. In view of the fact that most of the non-Newtonian fluids have large Prandtl numbers, this study was directed toward such fluids. The effects of the porous medium parameters, k1 and k2, body shape parameter, m, and surface thermal variations parameter, p, as well as the power-law index, n, were examined. © 2010 Springer Science+Business Media B.V.

  8. Non-Darcy Free Convection of Power-Law Fluids Over a Two-Dimensional Body Embedded in a Porous Medium

    KAUST Repository

    El-Amin, Mohamed; Sun, Shuyu; El-Ameen, M. A.; Jaha, Y. A.; Gorla, Rama Subba Reddy

    2010-01-01

    A boundary layer analysis was presented to study the non-Darcy-free convection of a power-law fluid over a non-isothermal two-dimensional body embedded in a porous medium. The Ostwald-de Waele power-law model was used to characterize the non-Newtonian fluid behavior. Similarity solutions were obtained with variations in surface temperature or surface heat flux. In view of the fact that most of the non-Newtonian fluids have large Prandtl numbers, this study was directed toward such fluids. The effects of the porous medium parameters, k1 and k2, body shape parameter, m, and surface thermal variations parameter, p, as well as the power-law index, n, were examined. © 2010 Springer Science+Business Media B.V.

  9. Serum Thyroxine to Thyroxine-Binding Globulin Ratio in Pregnancy and Newborn

    International Nuclear Information System (INIS)

    Kim, Ji Yeul

    1982-01-01

    To evaluate the diagnostic value of the ratio of serum thyroxine(T 4 ) /thyroxine-binding globulin (TBG) for the thyroid status in pregnancy and newborn serum thyroxine, TBG, triiodothyronine, and free thyroxine levels were radioimmunoassayed in normal pregnant women at each of the trimesters, and the calculated serum T 4 /TBG ratios were compared with other parameters such as T 3 /TBG ratio and free T 4 /TBG ratio and free T 4 /TBG ratio. Serum T 4 levels were elevated with the proportionate increase in TBG levels during pregnancy, leading to the nearly constant value of serum T 4 /TBG ratios as in normal non-pregnant controls. In contrast, serum T 3 /TBG and free T 4 /TBG ratios varied considerably during pregnancy. In newborn, T 4 levels were nearly not changed with compared non-pregnant control value and TBG levels were elevated. The results indicate that serum T 4 /TBG ratio is a better parameter than others in evaluating the thyroid status during pregnancy and but newborn is a no better.

  10. Freeze Protection in Gas Holders

    DEFF Research Database (Denmark)

    Hjorth, Poul G.; Duursma, Gail

    In cold weather, the water seals of gasholders need protection from freez- ing to avoid compromising the seal. These holders have a large reservoir of “tank water” at the base which is below ground. At present freeze- protection is achieved by external heating of the seal water which...

  11. Fasting induces the generation of serum thyronine-binding globulin in Zucker rats

    International Nuclear Information System (INIS)

    Young, R.A.; Rajatanavin, R.; Moring, A.F.; Braverman, L.E.

    1985-01-01

    Five-month-old lean and obese Zucker rats were fasted for up to 7 days (lean rats) or 28 days (obese rats), and serum total and free T4 and T3 concentrations, percent free T4 and T3 by equilibrium dialysis, and the binding of [ 125 I] T4 to serum proteins by gel electrophoresis were measured. In the lean rats, a 4- or 7-day fast resulted in significant decreases in serum total and free T4 and T3 concentrations. There was a decrease in the percent free T3 after 7 days of starvation. In contrast, a 4- or 7-day fast did not alter any of these variables in the obese rats. However, after 14 or more days of starvation, serum total T4 and T3 concentrations increased, and the percent free T4 and T3 decreased, resulting in no change in the serum free T4 or T3 concentrations in the obese rats. The percent of [ 125 I]T4 bound to serum thyronine-binding globulin increased and the percent bound to thyronine-binding prealbumin decreased with the duration of the fast in both the lean and obese rats. The increase in serum thyronine-binding globulin binding of T4 can explain the increase in serum total T4 and T3 concentrations, the decrease in percent free T4 and T3, and the normal free hormone concentration in the long term fasted obese rats. The findings in the lean rats appear to be due to a combination of the known central hypothyroidism that occurs during 4-7 days of fasting and the fasting-induced changes in T4 binding in serum. Changes in T4 and T3 binding in serum during fasting in the rat must be considered when the effects of fasting on serum concentrations of the thyroid hormones, thyroid hormone kinetics, and the peripheral action of the thyroid hormones are evaluated

  12. Freeze dehydration of milk using microwave energy

    International Nuclear Information System (INIS)

    Souda, K.B.; Akyel, C.; Bilgen, E.

    1989-01-01

    This paper presents the results of experimental studies on heat and mass transfer during a microwave freeze dehydration process. An experimental system and procedure was developed to freeze dry milk. A 2500-W microwave system with an appropriate wave guide was set up and instrumented, and a procedure was experimentally developed to obtain milk powder first by freezing milk and then dehydrating it at low pressure using microwave energy. An unsteady-state analysis was used to derive a one-dimensional mathematical model of the freeze dehydration process in a microwave electromagnetic field

  13. Microencapsulation by freeze-drying of potassium norbixinate and curcumin with maltodextrin: stability, solubility, and food application.

    Science.gov (United States)

    Sousdaleff, Mirian; Baesso, Mauro Luciano; Medina Neto, Antonio; Nogueira, Ana Cláudia; Marcolino, Vanessa Aparecida; Matioli, Graciette

    2013-01-30

    Stability of potassium norbixinate and curcumin by microencapsulation with maltodextrin DE20 and freeze-drying was evaluated as a function of exposition to light, air, different pH, water solubility, and in food applications. The best results were obtained with microencapsulated potassium norbixinate 1:20, which, when vacuum-packed and in the presence of natural light, showed color retention of 78%, while microencapsulated curcumin 1:20 showed color retention of 71%. Differential scanning calorimetry and thermogravimetry provided an indication of interaction between colorants and maltodextrin. Photoacoustic spectroscopy (PAS) showed that free and microencapsulated colorants exhibited high rates of absorption throughout the measured spectral region. This work evidenced that the freeze-drying process is favorable for microencapsulation of curcumin by maltodextrin, providing improved solubility to the microencapsulated colorant. Both microencapsulated colorants showed relevant results for use in a wide range of pH and food applications. The PAS technique was useful for the evaluation of the stability of free and microencapsulated colorants.

  14. Freeze-Thaw Durability of Air-Entrained Concrete

    Directory of Open Access Journals (Sweden)

    Huai-Shuai Shang

    2013-01-01

    Full Text Available One of the most damaging actions affecting concrete is the abrupt temperature change (freeze-thaw cycles. The types of deterioration of concrete structures by cyclic freeze-thaw can be largely classified into surface scaling (characterized by the weight loss and internal crack growth (characterized by the loss of dynamic modulus of elasticity. The present study explored the durability of concrete made with air-entraining agent subjected to 0, 100, 200, 300, and 400 cycles of freeze-thaw. The experimental study of C20, C25, C30, C40, and C50 air-entrained concrete specimens was completed according to “the test method of long-term and durability on ordinary concrete” GB/T 50082-2009. The dynamic modulus of elasticity and weight loss of specimens were measured after different cycles of freeze-thaw. The influence of freeze-thaw cycles on the relative dynamic modulus of elasticity and weight loss was analyzed. The findings showed that the dynamic modulus of elasticity and weight decreased as the freeze-thaw cycles were repeated. They revealed that the C30, C40, and C50 air-entrained concrete was still durable after 300 cycles of freeze-thaw according to the experimental results.

  15. Radioimmunological determination of reverse triiodo thyronine in unextracted serum and serum dialysates

    International Nuclear Information System (INIS)

    Laurberg, P.; Weeke, J.

    1977-01-01

    A sensitive radioimmunoassay for measurements of 3,3,5-triiodothyronine (reverse T3,rT3) in small amounts of unextracted serum is described. The interference of rT3 binding proteins in serum was precluded by addition of 8-anilinol-naphthalene sulphonic acid (ANS). The cross reaction of T4 with the rT3 anti-serum varied with the concentration of T4 in the samples. At 50 per cent inhibition of I 125 rT3 binding, the T4 cross reaction was 0.075% (mol/mol). All values were corrected for T4 cross reaction. By the present method total rT3 averaged 0.37 nmol/l in thirty-two normal subjects. Higher values (0.81-1.98 nmol/l) were obtained in seventeen thyrotoxic patients, while the rT3 was very low (<=0.046 nmol/l) in ten patients with severe primary hypothyroidism. A modification of the total rT3 assay was used for measurements of rT3 in serum dialysates (free rT3). The sensitivity was 0.46 pmol/l. This sensitivity did not allow detection of free rT3 in all normal subjects. (Auth.)

  16. Effects of the free fatty acid content in yellow grease on performance, carcass characteristics, and serum lipids in broilers.

    Science.gov (United States)

    Wu, H; Gong, L M; Guo, L; Zhang, L Y; Li, J T

    2011-09-01

    This study was conducted to investigate whether the free fatty acid (FFA) content of yellow grease would influence the performance and carcass characteristics of broiler chicks. A total of 432 one-day-old, male Arbor Acres broilers were randomly allotted to 1 of 4 treatments, with each treatment being applied to 6 pens of 18 chicks. The dietary treatments were based on corn and soybean meal and were supplemented with either soybean oil (2.86% FFA) or yellow grease with a low (2.74%), medium (12.59%), or high (19.05%) FFA content. The fat sources were supplemented at 1.5% of the diet during the starter phase (0 to 21 d) and at 3.0% of the diet during the grower phase (22 to 42 d). From d 0 to 42, the BW gains of chicks fed diets containing soybean oil and yellow grease with 2.74% FFA were similar. As the FFA level in the yellow grease increased, the BW gain of chicks decreased (P dressing percentage and the percentage of breast muscle in the carcass were significantly (P < 0.01) lower for broilers fed any yellow grease diet compared with birds fed soybean oil. In contrast, abdominal fat was significantly increased in diets containing yellow grease. These results demonstrate that the performance of birds fed yellow grease with a low content of FFA was essentially equal to that of birds fed soybean oil. However, because yellow grease samples containing FFA levels greater than 2.74% negatively affected bird performance, producers should exercise caution with regard to feeding broilers yellow grease with an elevated FFA content. In 42-d-old broilers, serum total cholesterol and low-density lipoprotein cholesterol levels were elevated in birds fed yellow grease, regardless of the dietary level. In contrast, serum high-density lipoprotein cholesterol and triglyceride levels were unaffected by dietary treatment. Although dietary FFA may influence triglyceride-rich lipoprotein metabolism in broilers, an explanation for the observed effects remains elusive.

  17. Effect of chemical stabilizers on the thermostability and infectivity of a representative panel of freeze dried viruses.

    Directory of Open Access Journals (Sweden)

    Boris Pastorino

    Full Text Available As a partner of the European Virus Archive (EVA FP7 project, our laboratory maintains a large collection of freeze-dried viruses. The distribution of these viruses to academic researchers, public health organizations and industry is one major aim of the EVA consortium. It is known that lyophilization requires appropriate stabilizers to prevent inactivation of the virus. However, few studies have investigated the influence of different stabilizers and lyophilization protocols on the thermostability of different viruses. In order to identify optimal lyophilization conditions that will deliver maximum retention of viral infectivity titre, different stabilizer formulations containing trehalose, sorbitol, sucrose or foetal bovine serum were evaluated for their efficacy in stabilizing a representative panel of freeze dried viruses at different storage temperatures (-20°C, +4°C and +20°C for one week, the two latter mimicking suboptimal shipping conditions. The Tissue Culture Infectious Dose 50% (TCID50 assay was used to compare the titres of infectious virus. The results obtained using four relevant and model viruses (enveloped/non enveloped RNA/DNA viruses still serve to improve the freeze drying conditions needed for the development and the distribution of a large virus collection.

  18. A Theory of Immersion Freezing

    Science.gov (United States)

    Barahona, Donifan

    2017-01-01

    Immersion freezing is likely involved in the initiation of precipitation and determines to large extent the phase partitioning in convective clouds. Theoretical models commonly used to describe immersion freezing in atmospheric models are based on the classical nucleation theory which however neglects important interactions near the immersed particle that may affect nucleation rates. This work introduces a new theory of immersion freezing based on two premises. First, immersion ice nucleation is mediated by the modification of the properties of water near the particle-liquid interface, rather than by the geometry of the ice germ. Second, the same mechanism that leads to the decrease in the work of germ formation also decreases the mobility of water molecules near the immersed particle. These two premises allow establishing general thermodynamic constraints to the ice nucleation rate. Analysis of the new theory shows that active sites likely trigger ice nucleation, but they do not control the overall nucleation rate nor the probability of freezing. It also suggests that materials with different ice nucleation efficiency may exhibit similar freezing temperatures under similar conditions but differ in their sensitivity to particle surface area and cooling rate. Predicted nucleation rates show good agreement with observations for a diverse set of materials including dust, black carbon and bacterial ice nucleating particles. The application of the new theory within the NASA Global Earth System Model (GEOS-5) is also discussed.

  19. Utility of free prostate specific antigen serum level and its related parameters in the diagnosis of prostate cancer

    Directory of Open Access Journals (Sweden)

    Azmi A Haroun

    2011-01-01

    Full Text Available We evaluated the role of free prostate specific antigen (f-PSA serum level and its related parameters in detecting prostate cancer. This retrospective study was conducted between January 2006 and March 2008. Trans-rectal ultrasound guided prostate biopsy was performed for 107 patients who had total PSA (t-PSA level of either >4 ng/mL with or without palpable nodule or ≤4 ng/mL with palpable nodule on digital rectal examination. The perfor-mance measurements for f-PSA, percent free PSA (%f-PSA and free PSA density (f-PSAD were determined and compared with those for t-PSA and total PSA density (t-PSAD. Descriptive statistics for all variables of interest were calculated, and receiver operating characteristic curves were generated. Nine patients (8.4% had normal histology, 69 patients (64.4% had benign disease and 29 patients (27.1% had prostate cancer. The performance of f-PSA in PCa detection was better than other evaluated parameters. The largest area under the curve for patients in the gray area (t-PSA range 4.1-10 ng/mL was for f-PSA, with a value of 0.64 and a sensitivity and specificity of 44% and 87%, respectively. For %f-PSA, these values were 0.59, 63% and 62%, respectively. For patients with a t-PSA level of 10.1-20 ng/mL, they were 0.68, 67%, and 81%, respectively, for f-PSA, and 0.64, 67%, and 76%, respectively, for %f-PSA. In conclusion, f-PSA serum levels performed better than free to total PSA ratio and t-PSA for prostate cancer screening. It is of clinical value which could affect the biopsy decision avoiding unnecessary interventions.

  20. Vial freeze-drying, part 1: new insights into heat transfer characteristics of tubing and molded vials.

    Science.gov (United States)

    Hibler, Susanne; Wagner, Christophe; Gieseler, Henning

    2012-03-01

    In order to optimize a freeze-drying cycle, information regarding the heat transfer characteristics of the container system is imperative. Two most recently developed tubing (TopLyo™) and molded (EasyLyo™) vial designs were compared with a standard serum tubing and molded vial, a polymer vial (TopPac™), and an amber molded EasyLyo™. In addition, the impact of methodology on the determination of reliable vial heat transfer coefficient (K(v) ) data is examined in detail. All K(v) s were gravimetrically determined by sublimation tests with pure water at 50, 100, 200, and 400 mTorr. In contrast to the traditional assumption that molded vials exhibit inefficient heat transfer characteristics, these vials showed a very similar performance compared with their serum tubing counterparts in the relevant pressure range for freeze-drying. At 100 mTorr, the TopLyo™ center vials show only 4% higher K(v) values than the EasyLyo™ center vials. All glass vials outmatch the polymer vial in terms of heat transfer, up to 30% elevated heat transfer for the TopLyo™ center vials at 400 mTorr. Sublimation tests have demonstrated to be a valuable tool to investigate the heat transfer characteristics of vials, but results are dependent on methodology. New developments in molded vial manufacturing lead to improved heat transfer performance. Copyright © 2011 Wiley Periodicals, Inc.

  1. Freezing and extreme-value statistics in a random energy model with logarithmically correlated potential

    International Nuclear Information System (INIS)

    Fyodorov, Yan V; Bouchaud, Jean-Philippe

    2008-01-01

    We investigate some implications of the freezing scenario proposed by Carpentier and Le Doussal (CLD) for a random energy model (REM) with logarithmically correlated random potential. We introduce a particular (circular) variant of the model, and show that the integer moments of the partition function in the high-temperature phase are given by the well-known Dyson Coulomb gas integrals. The CLD freezing scenario allows one to use those moments for extracting the distribution of the free energy in both high- and low-temperature phases. In particular, it yields the full distribution of the minimal value in the potential sequence. This provides an explicit new class of extreme-value statistics for strongly correlated variables, manifestly different from the standard Gumbel class. (fast track communication)

  2. Freezing and extreme-value statistics in a random energy model with logarithmically correlated potential

    Energy Technology Data Exchange (ETDEWEB)

    Fyodorov, Yan V [School of Mathematical Sciences, University of Nottingham, Nottingham NG72RD (United Kingdom); Bouchaud, Jean-Philippe [Science and Finance, Capital Fund Management 6-8 Bd Haussmann, 75009 Paris (France)

    2008-09-19

    We investigate some implications of the freezing scenario proposed by Carpentier and Le Doussal (CLD) for a random energy model (REM) with logarithmically correlated random potential. We introduce a particular (circular) variant of the model, and show that the integer moments of the partition function in the high-temperature phase are given by the well-known Dyson Coulomb gas integrals. The CLD freezing scenario allows one to use those moments for extracting the distribution of the free energy in both high- and low-temperature phases. In particular, it yields the full distribution of the minimal value in the potential sequence. This provides an explicit new class of extreme-value statistics for strongly correlated variables, manifestly different from the standard Gumbel class. (fast track communication)

  3. Comparison of blood serum selenium concentrations from elderly population living in Sao Paulo city and literature data

    International Nuclear Information System (INIS)

    Saiki, Mitiko; Alves, Edson R.; Vasconcellos, Marina B.A.; Sumita, Nairo M.; Jaluul, Omar; Jacob Filho, Wilson

    2005-01-01

    Selenium is an essential nutrient of fundamental importance to human biology. This element is a component of the enzyme glutathione peroxidase that avoids the formation of free radicals protecting the organism against the oxidative damage. The Se concentrations in serum vary from one part of the world to another and this fact has been satisfactorily explained as a consequence of different Se intakes of the population. The main objective of this paper was the determination of serum Se concentrations in healthy elderly population included in a program for 'Successful Ageing' of the Hospital das Clinicas, Sao Paulo University Medical School. The blood samples were collected after a 12 h fast from 32 subjects aged 60 to 87 years. The blood was centrifuged after completely clotted and 4.0 mL of serum were freeze.dried for Se determination using neutron activation analysis. The serum samples and Se standard were irradiated for 16 h under a thermal neutron flux of de 5 x 10 12 n cm - '2 s -1 at the IEA-R1 nuclear reactor. Se was determined by measuring the gamma activity of 75 Se using an HGe detector coupled to a gamma ray spectrometer. The mean value of (0.92±0.07) μmol L -1 was obtained for serum Se concentration. This value is within the data used as reference values in clinical laboratories and those published in literature. Results obtained for certified reference materials NIST 1566b Oyster Tissue and NIST 1577b Bovine Liver showed good precision and agreed with the certified values. The relative standard deviation of the results was lower than 4.8 % and relative error lower than 0.97 %. Detection limit value obtained for serum Se determination was 0.003 μmol L-1. (author)

  4. Medium corrections to nucleon-nucleon interactions

    International Nuclear Information System (INIS)

    Dortmans, P.J.; Amos, K.

    1990-01-01

    The Bethe-Goldstone equations have been solved for both negative and positive energies to specify two nucleon G-matrices fully off of the energy shell. Medium correction effects of Pauli blocking and of the auxiliary potential are included in infinite matter systems characterized by fermi momenta in the range 0.5 fm -1 to 1.8 fm -1 . The Paris interaction is used as the starting potential in most calculations. Medium corrections are shown to be very significant over a large range of energies and densities. On the energy shell values of G-matrices vary markedly from those of free two nucleon (NN) t-matrices which have been solved by way of the Lippmann-Schwinger equation. Off of the energy shell, however, the free and medium corrected Kowalski-Noyes f-ratios rate are quite similar suggesting that a useful model of medium corrected G-matrices are appropriately scaled free NN t-matrices. The choice of auxiliary potential form is also shown to play a decisive role in the negative energy regime, especially when the saturation of nuclear matter is considered. 30 refs., 7 tabs., 7 figs

  5. New permeable cryoprotectant-free vitrification method for native human sperm.

    Science.gov (United States)

    Aizpurua, J; Medrano, L; Enciso, M; Sarasa, J; Romero, A; Fernández, M A; Gómez-Torres, M J

    2017-10-01

    Is permeable cryoprotectant-free vitrification of native sperm samples a good alternative to conventional slow freezing? The permeable cryoprotectant-free sperm vitrification protocol tested in this study renders considerably better recovery rates of good quality sperm compared to slow freezing. Slow freezing is currently the most commonly used technique for sperm cryopreservation, though this method has been repeatedly shown to have negative effects on both structural and functional sperm features. New alternative methods such as vitrification have been established as a successful alternative in other reproductive cell types, but vitrification of spermatozoa is still a rather unexplored methodology, with limited studies showing its efficacy in male gametes. This study included 18 normozoospermic sperm samples from patients seeking ART treatment between 2014 and 2015. The effects of a new vitrification protocol on functional and structural sperm quality parameters in comparison to fresh and slow-frozen samples were assessed. All samples were divided into three aliquots: fresh (F), slow freezing-thawing (S) and vitrification-warming (V). Sperm concentration, motility, morphology, vitality, DNA fragmentation, cytoskeleton integrity and spontaneous acrosome reaction were assessed and compared between the groups. Results showed improved preservation of sperm features after vitrification compared to conventional freezing. Permeable cryoprotectant-free vitrification presented a significantly higher percentage of live spermatozoa, than slow freezing, better preservation of acrosomes was achieved in vitrified samples and DNA fragmentation was reduced approximately one-third on average compared to slow freezing. Regarding tubulin assay, three different labelling patterns were observed. The frequency of these labelling patterns was similar in F and V groups but this was not the case of the S group. The multivariate analysis of all sperm quality parameters studied revealed

  6. COMPARATIVE STUDIES ON EFFECTS OF FREEZING ON PHYSICOCHEMICAL PROPERTIES OF FILLETS TWO FISH SPECIES IN IRAN

    Directory of Open Access Journals (Sweden)

    Ali Aberoumand

    2018-04-01

    Full Text Available Because of fishes Sparidentex hasta and Pampus argenteus in the southern of Iran are consumed abundant in a particular season and it should be frozen for consumption throughout the year. Therefore, this research was carried out to investigate the effects of freezing on some of the physicochemical properties of fillets the fishes. Factors such as fat with chloroform-methanol method, amount of TBA (Thiobarbituric acid in fish muscle accordance method of Pearson, pH using a pH meter, FFA (Free fatty acids with titration in the presence of phenolphthalein was determined based on the percentage of oleic acid, peroxide value according to AOAC (Association of Official Analytical Chemists, in fresh samples at time zero and after different periods of freezing were tested respectively. Result showd that TBA (Thiobarbituric acid content in fish fillet, found Pampus argenteus and Sparidentex hasta 0.65 and 0.53 respectively. The results showed that the highest percentage of fat found for Pampus argenteus at 95 days 24.2(%2 and for Sparidentex hasta at 35 days (25.19%, free fatty acids contents found highest (0.9% and (0.97% for Sparidentex hasta and Pampus argenteus after 95 days. It can conclude that the TBA (Thiobarbituric acid, FFA (Free fatty acids  contents and pH of both fish species during storage in freezer were increased. Peroxide value in fish Pampus argenteus was reduced but, in Sparidentex hasta showed no significant differently.  The best time of storage of fishes Pampus argenteus and Sparidentex hasta at -18 °C was 35 days freezing, but nutritional value of fillets and fatty acids greatly reduced

  7. Transient transfection of serum-free suspension HEK 293 cell culture for efficient production of human rFVIII

    Science.gov (United States)

    2011-01-01

    Background Hemophilia A is a bleeding disorder caused by deficiency in coagulation factor VIII. Recombinant factor VIII (rFVIII) is an alternative to plasma-derived FVIII for the treatment of hemophilia A. However, commercial manufacturing of rFVIII products is inefficient and costly and is associated to high prices and product shortage, even in economically privileged countries. This situation may be solved by adopting more efficient production methods. Here, we evaluated the potential of transient transfection in producing rFVIII in serum-free suspension HEK 293 cell cultures and investigated the effects of different DNA concentration (0.4, 0.6 and 0.8 μg/106 cells) and repeated transfections done at 34° and 37°C. Results We observed a decrease in cell growth when high DNA concentrations were used, but no significant differences in transfection efficiency and in the biological activity of the rFVIII were noticed. The best condition for rFVIII production was obtained with repeated transfections at 34°C using 0.4 μg DNA/106 cells through which almost 50 IU of active rFVIII was produced six days post-transfection. Conclusion Serum-free suspension transient transfection is thus a viable option for high-yield-rFVIII production. Work is in progress to further optimize the process and validate its scalability. PMID:22115125

  8. Culture medium type affects endocytosis of multi-walled carbon nanotubes in BEAS-2B cells and subsequent biological response.

    Science.gov (United States)

    Haniu, Hisao; Saito, Naoto; Matsuda, Yoshikazu; Tsukahara, Tamotsu; Maruyama, Kayo; Usui, Yuki; Aoki, Kaoru; Takanashi, Seiji; Kobayashi, Shinsuke; Nomura, Hiroki; Okamoto, Masanori; Shimizu, Masayuki; Kato, Hiroyuki

    2013-09-01

    We examined the cytotoxicity of multi-walled carbon nanotubes (MWCNTs) and the resulting cytokine secretion in BEAS-2B cells or normal human bronchial epithelial cells (HBEpCs) in two types of culture media (Ham's F12 containing 10% FBS [Ham's F12] and serum-free growth medium [SFGM]). Cellular uptake of MWCNT was observed by fluorescent microscopy and analyzed using flow cytometry. Moreover, we evaluated whether MWCNT uptake was suppressed by 2 types of endocytosis inhibitors. We found that BEAS-2B cells cultured in Ham's F12 and HBEpCs cultured in SFGM showed similar biological responses, but BEAS-2B cells cultured in SFGM did not internalize MWCNTs, and the 50% inhibitory concentration value, i.e., the cytotoxicity, was increased by more than 10-fold. MWCNT uptake was suppressed by a clathrin-mediated endocytosis inhibitor and a caveolae-mediated endocytosis inhibitor in BEAS-2B cells cultured in Ham's F12 and HBEpCs cultured in SFGM. In conclusion, we suggest that BEAS-2B cells cultured in a medium containing serum should be used for the safety evaluation of nanomaterials as a model of normal human bronchial epithelial cells. However, the culture medium composition may affect the proteins that are expressed on the cytoplasmic membrane, which may influence the biological response to MWCNTs. Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.

  9. Freezing tolerance of conifer seeds and germinants.

    Science.gov (United States)

    Hawkins, B J; Guest, H J; Kolotelo, D

    2003-12-01

    Survival after freezing was measured for seeds and germinants of four seedlots each of interior spruce (Picea glauca x engelmannii complex), lodgepole pine (Pinus contorta Dougl. ex Loud.), Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and western red cedar (Thuja plicata Donn ex D. Donn). Effects of eight seed treatments on post-freezing survival of seeds and germinants were tested: dry, imbibed and stratified seed, and seed placed in a growth chamber for 2, 5, 10, 15, 20 or 30 days in a 16-h photoperiod and a 22/17 degrees C thermoperiod. Survival was related to the water content of seeds and germinants, germination rate and seedlot origin. After freezing for 3 h at -196 degrees C, dry seed of most seedlots of interior spruce, Douglas-fir and western red cedar had 84-96% germination, whereas lodgepole pine seedlots had 53-82% germination. Freezing tolerance declined significantly after imbibition in lodgepole pine, Douglas-fir and interior spruce seed (western red cedar was not tested), and mean LT50 of imbibed seed of these species was -30, -24.5 and -20 degrees C, respectively. Freezing tolerance continued to decline to a minimum LT50 of -4 to -7 degrees C after 10 days in a growth chamber for interior spruce, Douglas-fir and lodgepole pine, or after 15 days for western red cedar. Minimum freezing tolerance was reached at the stage of rapid hypocotyl elongation. In all species, a slight increase in freezing tolerance of germinants was observed once cotyledons emerged from the seed coat. The decrease in freezing tolerance during the transition from dry to germinating seed correlated with increases in seed water content. Changes in freezing tolerance between 10 and 30 days in the growth chamber were not correlated with seedling water content. Within a species, seedlots differed significantly in freezing tolerance after 2 or 5 days in the growth chamber. Because all seedlots of interior spruce and lodgepole pine germinated quickly, there was no correlation

  10. Physical Stability of Freeze-Dried Isomalt Diastereomer Mixtures

    DEFF Research Database (Denmark)

    Koskinen, Anna-Kaisa; Fraser-Miller, Sara J.; Bøtker, Johan P.

    2016-01-01

    Purpose Isomalt is a sugar alcohol used as an excipient in commercially available solid oral dosage forms. The potential of isomalt as a novel freeze-drying excipient was studied in order to increase knowledge of the behavior of isomalt when it is freeze-dried. Methods Isomalt was freeze-dried in......Purpose Isomalt is a sugar alcohol used as an excipient in commercially available solid oral dosage forms. The potential of isomalt as a novel freeze-drying excipient was studied in order to increase knowledge of the behavior of isomalt when it is freeze-dried. Methods Isomalt was freeze......-dried in four different diastereomer compositions and its physical stability was investigated with differential scanning calorimetry, Fourier-transform infrared and Raman spectroscopy, X-ray powder diffraction, Karl-Fischer titration and thermogravimetric analysis in order to verify the solid state form...... of the diastereomer compositions showed signs of physical instability when stored in the highest relative humidity condition. The four different crystalline diastereomer mixtures showed specific identifiable solid state properties. Conclusions Isomalt was shown to be a suitable excipient for freeze-drying. Preferably...

  11. SELDI-TOF MS of quadruplicate urine and serum samples to evaluate changes related to storage conditions.

    Science.gov (United States)

    Traum, Avram Z; Wells, Meghan P; Aivado, Manuel; Libermann, Towia A; Ramoni, Marco F; Schachter, Asher D

    2006-03-01

    Proteomic profiling with SELDI-TOF MS has facilitated the discovery of disease-specific protein profiles. However, multicenter studies are often hindered by the logistics required for prompt deep-freezing of samples in liquid nitrogen or dry ice within the clinic setting prior to shipping. We report high concordance between MS profiles within sets of quadruplicate split urine and serum samples deep-frozen at 0, 2, 6, and 24 h after sample collection. Gage R&R results confirm that deep-freezing times are not a statistically significant source of SELDI-TOF MS variability for either blood or urine.

  12. Freezing the Master Production Schedule Under Rolling Planning Horizons

    OpenAIRE

    V. Sridharan; William L. Berry; V. Udayabhanu

    1987-01-01

    The stability of the Master Production Schedule (MPS) is a critical issue in managing production operations with a Material Requirements Planning System. One method of achieving stability is to freeze some portion or all of the MPS. While freezing the MPS can limit the number of schedule changes, it can also produce an increase in production and inventory costs. This paper examines three decision variables in freezing the MPS: the freezing method, the freeze interval length, and the planning ...

  13. Comparison between human cord blood serum and platelet-rich plasma supplementation for Human Wharton's Jelly Stem Cells and dermal fibroblasts culture

    Directory of Open Access Journals (Sweden)

    Hashemi SS

    2016-08-01

    Full Text Available We carried out a side-by-side comparison of the effects of Human cord blood serum (HcbS versus embryonic PRP on Human Wharton's Jelly Stem Cells(hWMSCand dermal fibroblasts proliferation. Human umbilical cord blood was collected to prepare activated serum (HCS and platelet-rich plasma (CPRP.Wharton's Jelly Stem Cells and dermal fibroblasts were cultured in complete medium with10% CPRP, 10%HCSor 10% fetal bovine serumand control (serum-free media.The efficiency of the protocols was evaluated in terms of the number of adherent cells and their expansion and Cell proliferation. We showed that proliferation of fibroblasts and mesenchymal stem cells in the presence of cord blood serum and platelet-rich plasma significantly more than the control group (p≤0/05. As an alternative to FBS, cord blood serum has been proved as an effective component in cell tissue culture applications and embraced a vast future in clinical applications of regenerative medicine. However, there is still a need to explore the potential of HCS and its safe applications in humanized cell therapy or tissue engineering.

  14. Rapid formation of the 110 K phase in Bi-Pb-Sr-Ca-Cu-O through freeze-drying powder processing

    International Nuclear Information System (INIS)

    Song, K.H.; Liu, H.K.; Dou, S.X.; Sorrell, C.C.

    1990-01-01

    This paper reports three techniques for processing Bi-Pb-Sr-Ca-Cu-O (BPSCCO) powders investigated: dry-mixing, sol-gel formation, and freeze-drying. It was found that sintering for 120 h at 850 degrees C is required to form nearly single-phase (Bi,Pb) 2 Sr 2 Ca 2 Cu 3 O 10-y by dry-mixing, whereas sintering for 30 h at 840 degrees C was sufficient to form the 110 K (2223) phase when freeze-drying was used. The sol-gel route was found to be intermediate in efficiency between these two techniques. Freeze-drying provided highly reactive, intimately mixed, and carbon-free precursors. The presence of carbonates in the uncalcined powders was the major cause of phase segregation and sluggishness of the 110 K phase formation

  15. Experimental determination of the key heat transfer mechanisms in pharmaceutical freeze-drying.

    Science.gov (United States)

    Ganguly, Arnab; Nail, Steven L; Alexeenko, Alina

    2013-05-01

    The study is aimed at quantifying the relative contribution of key heat transfer modes in lyophilization. Measurements of vial heat transfer rates in a laboratory-scale freeze-dryer were performed using pure water, which was partially sublimed under various conditions. The separation distance between the shelf and the vial was systematically varied, and sublimation rates were determined gravimetrically. The heat transfer rates were observed to be independent of separation distance between the vial and the shelf and linearly dependent on pressure in the free molecular flow limit, realized at low pressures (120 mTorr), heat transfer rates were independent of pressure and inversely proportional to separation distance. Previous heat transfer studies in conventional freeze-drying cycles have attributed a dominant portion of the total heat transfer to radiation, the rest to conduction, whereas convection has been found to be insignificant. Although the measurements reported here confirm the significance of the radiative and gas conduction components, the convective component has been found to be comparable to the gas conduction contribution at pressures greater than 100 mTorr. The current investigation supports the conclusion that the convective component of the heat transfer cannot be ignored in typical laboratory-scale freeze-drying conditions. Copyright © 2013 Wiley Periodicals, Inc.

  16. Chloride Transport in OPC Concrete Subjected to the Freeze and Thaw Damage

    Directory of Open Access Journals (Sweden)

    Ki Yong Ann

    2017-01-01

    Full Text Available To predict the durability of a concrete structure under the coupling degradation consisting of the frosting and chloride attack, microstructural analysis of the concrete pore structure should be accompanied. In this study, the correlation between the pore structure and chloride migration for OPC concrete was evaluated at the different cement content in the concrete mix accounting for 300, 350, and 400 kg/m3 at 0.45 of a free water cement ratio. The influence of frosting damage on the rate of chloride transport was assessed by testing with concrete specimens subjected to a rapid freezing and thawing cyclic environment. As a result, it was found that chloride transport was accelerated by frost damage, which was more influential at the lower cement content. The microscopic examination of the pore structure showed that the freezing environment increased the volume of the large capillary pore in the concrete matrix.

  17. Human amniotic fluid stem cells (hAFSCs expressing p21 and cyclin D1 genes retain excellent viability after freezing with (dimethyl sulfoxide DMSO

    Directory of Open Access Journals (Sweden)

    Shiva Gholizadeh-Ghaleh Aziz

    2018-04-01

    Full Text Available Human amniotic fluid stem cells (hAFSCs have features intermediate between embryonic and adult SCs, can differentiate into lineages of all three germ layers, and do not develop into tumors in vivo. Moreover, hAFSCs can be easily obtained in routine procedures and there is no ethical or legal limitations regarding their use for clinical and experimental applications. The aim of this study was to assess the effect of slow freezing/thawing and two different concentrations of DMSO (10% DMSO + 90% fetal bovine serum [FBS] and 5% DMSO + 95% FBS on the survival of hAFSCs. hAFSCs were obtained from 5 pregnant women during amniocentesis at 16–22 weeks of gestation. The expression of pluripotency markers (Octamer-binding transcription factor 4 [Oct4] and NANOG by reverse transcription polymerase chain reaction and cell surface markers (cluster of differentiation [CD31], CD44, CD45, and CD90 by flow cytometry was analyzed before and after the slow-freezing. Cell viability was assessed by trypan blue exclusion or MTT assay. Quantitative mRNA expression of Oct4, NANOG, cyclin D1 and p21 was determined by real-time PCR before and after the slow-freezing. Pluripotency of hAFSCs was confirmed by NANOG and POU5F1 (Oct4 gene expression before and after slow-freezing. All hAFSC cultures were positive for CD44 and CD90. A higher viability of hAFSCs was observed after freezing with 90% FBS + 10% DMSO. There was increased expression of NANOG and decreased expression of POU5F1 gene after freezing, compared to control cells (before freezing. DMSO and the process of freezing did not significantly change the expression of p21 and cyclin D1 genes in hAFSCs. Overall, our results indicate the applicability of slow-freezing and DMSO in cryopreservation of SCs.

  18. OPTIMIZATION OF MEDIUM COMPOSITIONS FOR SACCHAROMYCES BOULARDII BY BOX-BEHNKEN DESIGN

    Directory of Open Access Journals (Sweden)

    ZHANGTENG LEI

    2016-12-01

    Full Text Available Based on the previous experimental results, the Box-Behnken design (BBD was employed to study the individual and interactive effects of sucrose, malt extract, calf serum and sodium citrate on the growth of Saccharomyces boulardii, and the optimum medium compositions were obtained. Meanwhile, the optical density (OD in the fermentation suspension was measured at 560 nm after 36 h of incubation. It is shown that the optimized medium compositions were 36.28 g·L-1 sucrose, 6.38 g·L-1 malt extract, 5.69 g·L-1 calf serum and 5.3 g·L-1 sodium citrate. The result indicated that the growth of S. boulardii could increase significantly in the optimized medium, and the OD560nm value reached 1.397 ± 0.013 after 36 h, which increased 18.59 % compared with that of pre-optimized medium. In addition, the OD560nm value 1.397 ± 0.013 in the optimized medium was very closely to the expected value 1.394. This result suggested that optimization of medium compositions for S. boulardii by BBD in this study was reliable and effective.

  19. Determination of thymidine in serum used for cell culture media

    International Nuclear Information System (INIS)

    Schaer, J.C.; Maurer, U.; Schindler, R.

    1978-01-01

    Thymidine concentrations in serum used for cell culture media were determined with an assay based on isotope dilution. In this assay, incorporation of (3H)-thymidine into DNA of cultured cells was measured in the presence of 5 and 20% serum as a function of the concentration of unlabeled thymidine added to the medium. Thymidine concentrations were measured using horse serum as well as fetal calf serum in the culture media. Dialysis of serum resulted in a reduction of thymidine levels by factors of at least 10

  20. Comparing contact and immersion freezing from continuous flow diffusion chambers

    Directory of Open Access Journals (Sweden)

    B. Nagare

    2016-07-01

    Full Text Available Ice nucleating particles (INPs in the atmosphere are responsible for glaciating cloud droplets between 237 and 273 K. Different mechanisms of heterogeneous ice nucleation can compete under mixed-phase cloud conditions. Contact freezing is considered relevant because higher ice nucleation temperatures than for immersion freezing for the same INPs were observed. It has limitations because its efficiency depends on the number of collisions between cloud droplets and INPs. To date, direct comparisons of contact and immersion freezing with the same INP, for similar residence times and concentrations, are lacking. This study compares immersion and contact freezing efficiencies of three different INPs. The contact freezing data were obtained with the ETH CoLlision Ice Nucleation CHamber (CLINCH using 80 µm diameter droplets, which can interact with INPs for residence times of 2 and 4 s in the chamber. The contact freezing efficiency was calculated by estimating the number of collisions between droplets and particles. Theoretical formulations of collision efficiencies gave too high freezing efficiencies for all investigated INPs, namely AgI particles with 200 nm electrical mobility diameter, 400 and 800 nm diameter Arizona Test Dust (ATD and kaolinite particles. Comparison of freezing efficiencies by contact and immersion freezing is therefore limited by the accuracy of collision efficiencies. The concentration of particles was 1000 cm−3 for ATD and kaolinite and 500, 1000, 2000 and 5000 cm−3 for AgI. For concentrations  <  5000 cm−3, the droplets collect only one particle on average during their time in the chamber. For ATD and kaolinite particles, contact freezing efficiencies at 2 s residence time were smaller than at 4 s, which is in disagreement with a collisional contact freezing process but in accordance with immersion freezing or adhesion freezing. With “adhesion freezing”, we refer to a contact nucleation

  1. Biophysical evaluation of aminoclay as an effective protectant for protein stabilization during freeze-drying and storage

    Directory of Open Access Journals (Sweden)

    Song JG

    2016-12-01

    Full Text Available Jae Geun Song, Sang Hoon Lee, Hyo-Kyung Han College of Pharmacy, Dongguk University, Goyang, South Korea Abstract: This study aimed to evaluate aminoclay (3-aminopropyl-functionalized magnesium phyllosilicate as an effective protectant for the stabilization of protein formulation in freeze-drying. Bovine serum albumin (BSA, as a model protein, was freeze-dried with aminoclay at various concentrations, and the effects of aminoclay on the structural stability of proteins were compared with those of the conventional stabilizers. The structural characteristics of the protein were determined by size exclusion chromatography (SEC, circular dichroism (CD, and Fourier transform infrared (FTIR spectroscopy. Furthermore, physicochemical and morphological characteristics were examined by X-ray powder diffraction (XRPD, differential scanning calorimetry (DSC, and scanning electron microscopy (SEM. XRPD and DSC patterns indicated that the glass transition temperature (Tg of the amorphous formulation of aminoclay mixed with proteins was gradually elevated as the concentration of aminoclay increased. FTIR and CD spectral analysis suggested that the protein structure was well maintained with aminoclay during the freeze-drying process and 3 months of storage at 4°C and 40°C. Furthermore, aminoclay conferred the greatest protection against aggregation and retained the monomer content of BSA even at a high temperature. The morphological characteristics of lyophilized proteins were also well conserved during the storage with aminoclay. These results suggested that aminoclay may be useful as an alternative stabilizer for maintaining the structural stability of protein formulations. Keywords: aminoclay, cryoprotectant, lyoprotectant, freeze-drying, protein, stability

  2. 7 CFR 58.620 - Freezing and packaging rooms.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Freezing and packaging rooms. 58.620 Section 58.620 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards....620 Freezing and packaging rooms. The rooms used for freezing and packaging frozen desserts shall be...

  3. Physiological Mechanisms Only Tell Half Story: Multiple Biological Processes are involved in Regulating Freezing Tolerance of Imbibed Lactuca sativa Seeds.

    Science.gov (United States)

    Jaganathan, Ganesh K; Han, Yingying; Li, Weijie; Song, Danping; Song, Xiaoyan; Shen, Mengqi; Zhou, Qiang; Zhang, Chenxue; Liu, Baolin

    2017-03-13

    The physiological mechanisms by which imbibed seeds survive freezing temperatures in their natural environment have been categorized as freezing avoidance by supercooling and freezing tolerance by extracellular freeze-desiccation, but the biochemical and molecular mechanisms conferring seed freezing tolerance is unexplored. In this study, using imbibed Lactuca sativa seeds we show that fast cooled seeds (60 °C h -1 ) suffered significantly higher membrane damage at temperature between -20 °C and -10 °C than slow cooled (3 °Ch -1 ) seeds (P  0.05). However, both SOD activity and accumulation of free proline were induced significantly after slow cooling to -20 °C compared with fast cooling. RNA-seq demonstrated that multiple pathways were differentially regulated between slow and fast cooling. Real-time verification of some differentially expressed genes (DEGs) revealed that fast cooling caused mRNA level changes of plant hormone and ubiquitionation pathways at higher sub-zero temperature, whilst slow cooling caused mRNA level change of those pathways at lower sub-zero ttemperatures. Thus, we conclude that imbibed seed tolerate low temperature not only by physiological mechanisms but also by biochemical and molecular changes.

  4. Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support

    Directory of Open Access Journals (Sweden)

    Anita Muraglia

    2017-11-01

    Full Text Available Present cell culture medium supplements, in most cases based on animal sera, are not fully satisfactory especially for the in vitro expansion of cells intended for human cell therapy. This paper refers to (i an heparin-free human platelet lysate (PL devoid of serum or plasma components (v-PL and (ii an heparin-free human serum derived from plasma devoid of PL components (Pl-s and to their use as single components or in combination in primary or cell line cultures. Human mesenchymal stem cells (MSC primary cultures were obtained from adipose tissue, bone marrow, and umbilical cord. Human chondrocytes were obtained from articular cartilage biopsies. In general, MSC expanded in the presence of Pl-s alone showed a low or no proliferation in comparison to cells grown with the combination of Pl-s and v-PL. Confluent, growth-arrested cells, either human MSC or human articular chondrocytes, treated with v-PL resumed proliferation, whereas control cultures, not supplemented with v-PL, remained quiescent and did not proliferate. Interestingly, signal transduction pathways distinctive of proliferation were activated also in cells treated with v-PL in the absence of serum, when cell proliferation did not occur, indicating that v-PL could induce the cell re-entry in the cell cycle (cell commitment, but the presence of serum proteins was an absolute requirement for cell proliferation to happen. Indeed, Pl-s alone supported cell growth in constitutively activated cell lines (U-937, HeLa, HaCaT, and V-79 regardless of the co-presence of v-PL. Plasma- and plasma-derived serum were equally able to sustain cell proliferation although, for cells cultured in adhesion, the Pl-s was more efficient than the plasma from which it was derived. In conclusion, the cells expanded in the presence of the new additives maintained their differentiation potential and did not show alterations in their karyotype.

  5. Smoke-free policies and non-smokers’ reactions to SHS exposure in small and medium enterprises

    Directory of Open Access Journals (Sweden)

    Calliopi Sivri

    2013-12-01

    Full Text Available Introduction: Non-smoker employees can significantly improve the existing smoke-free policies in the workplace by asserting their right for smoke-free air and confronting smoker colleagues. The aim of the study was to assess the psychological and social drivers of non-smokers' readiness to assert their right for smoke-free air in the workplace. Materials and Methods: Twenty-six small-and-medium enterprises (SME with diverse background were randomly selected, and 284 employees agreed to participate in the study. Our study focused on the responses of 85 non-smokers (M age = 34 years, SD = 7.98, 84.2% worked in indoor offices. A cross-sectional design was used and participants completed a structured anonymous questionnaire assessing background and demographic characteristics, and psychosocial predictors of assertiveness intentions. Results: Although more than half of non-smokers reported they were often/almost always bothered by exposure to SHS, roughly one third of them reported having asked their colleagues not to smoke at work. Regression analysis showed that the effects of distal predictors (i.e. annoyance due to SHS exposure were mediated by past behaviour, attitudes (protection motivation beliefs, social norms, and self-efficacy. Conclusions: Health beliefs related to SHS exposure, and concerns about workplace health and job performance, social norms and self-efficacy can increase the assertiveness of non-smokers in workplace settings. Related campaigns should focus on communicating normative messages and self-efficacy training to empower non-smoker employees to act assertively towards protecting their smoke-free rights.

  6. Parameter Sensitivity of the Microdroplet Vacuum Freezing Process

    Directory of Open Access Journals (Sweden)

    Zhijun Zhang

    2015-01-01

    Full Text Available The vacuum freezing process of microdroplets (1 mm. Pressure and droplet diameter have an effect on cooling and freezing stages, but initial temperature only affects the cooling stage. The thermal conductivity coefficient kl affected the cooling stage, whereas ki affected the freezing stage. Heat capacity Cl affected the cooling stage, but Ci has virtually no effect on all stages. The actual latent heat of freezing ΔH was also affected. Higher density corresponds to lower cooling rate in the cooling stage.

  7. Isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of ten steroid metabolites in serum

    DEFF Research Database (Denmark)

    Søeborg, Tue; Frederiksen, Hanne; Johannsen, Trine Holm

    2017-01-01

    ), and estrone 3-sulfate (E1-S) in serum was developed and validated. Limits of quantification, variability (inter- and intra-day), analytical range and linearity were all found to be acceptable for clinical use. Furthermore, sample stability was evaluated including the influence of freeze-thaw cycles...

  8. Supersonic free jet, molecular free regime; Fondamenti fisici dei fasci molecolari supersonici. Parte 7. Il getto libero supersonico

    Energy Technology Data Exchange (ETDEWEB)

    Sanna, G. [ENEA, Centro Ricerche Frascati, Rome (Italy). Dipt. Innovazione; Tomassetti, G. [L' Aquila Univ., L' Aquila (Italy). Dipt. di Fisica

    1999-07-01

    The structure of the free jet emitted by a converging nozzle as obtained by the method of characteristics by Ashkenas e Sherman is described in details. In particular the dependence of the field variable by the distance from the nozzle is given. The transition from continuum to molecular free regime is then considered and the sudden freeze approximation is introduced. The processing of monoatomic and polyatomic gasses is also considered. [Italian] Le caratteristiche del campo di flusso in regime continuo di un getto libero supersonico ottenute col metodo delle caratteristiche da Ashekanas e Sherman sono messe in evidenza. In paricolare le relazioni analitiche per i parametri di flusso in funzione della distanza dal nozzle sono riportate. Viene poi considerata la transizione al regime molecolare ed e' introdotta la sudden freeze approximation. Vengono anche considerate le situazioni di non equilibrio tra gradi di liberta' interni ed esterni.

  9. Air-cooled steam condensers non-freeze warranties

    Energy Technology Data Exchange (ETDEWEB)

    Larinoff, M.W.

    1995-09-01

    What this paper is suggesting is the seller quote a condenser package with a LIMITED NON-FREEZE WARRANTY. Relieve the inexperienced buyer of the responsibility for selecting freeze protection design options. The seller cannot afford to over-design because of the added costs and the need for a competitive price. Yet he cannot under-design and allow the condenser tubes to freeze periodically and then pay the repair bills in accordance with the warranty.

  10. The unitary mechanism of infrared freezing in QCD with massive gluons

    International Nuclear Information System (INIS)

    Shirkov, D.V.

    1999-01-01

    A 'natural' model for the QCD invariant (running) coupling, free of the IR singularity, is proposed. It is based upon the hypothesis of finite gluon mass m gl existence and, technically, uses an accurate treating of threshold behaviour of Feynman diagram contribution. The model correlates with the unitarity condition. Quantitative estimate, performed in the one-loop approximation yields a reasonable lower bound for this mass m gl > 150 MeV and a smooth IR freezing at the level α s (Q 2 ) < or ∼ 1

  11. Microencapsulation of Purified Amylase Enzyme from Pitaya (Hylocereus polyrhizus Peel in Arabic Gum-Chitosan using Freeze Drying

    Directory of Open Access Journals (Sweden)

    Mehrnoush Amid

    2014-03-01

    Full Text Available Amylase is one of the most important enzymes in the world due to its wide application in various industries and biotechnological processes. In this study, amylase enzyme from Hylocereus polyrhizus was encapsulated for the first time in an Arabic gum-chitosan matrix using freeze drying. The encapsulated amylase retained complete biocatalytic activity and exhibited a shift in the optimum temperature and considerable increase in the pH and temperature stabilities compared to the free enzyme. Encapsulation of the enzyme protected the activity in the presence of ionic and non-ionic surfactants and oxidizing agents (H2O2 and enhanced the shelf life. The storage stability of amylase is found to markedly increase after immobilization and the freeze dried amylase exhibited maximum encapsulation efficiency value (96.2% after the encapsulation process. Therefore, the present study demonstrated that the encapsulation of the enzyme in a coating agent using freeze drying is an efficient method to keep the enzyme active and stable until required in industry.

  12. Fabrication of Li{sub 2}TiO{sub 3} pebbles by a freeze drying process

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Sang-Jin, E-mail: lee@mokpo.ac.kr [Department of Advanced Materials Science and Engineering, Mokpo National University, Muan 534-729 (Korea, Republic of); Park, Yi-Hyun [National Fusion Research Institute, Daejeon 305-806 (Korea, Republic of); Yu, Min-Woo [Department of Advanced Materials Science and Engineering, Mokpo National University, Muan 534-729 (Korea, Republic of)

    2013-11-15

    Li{sub 2}TiO{sub 3} pebbles were successfully fabricated by using a freeze drying process. The Li{sub 2}TiO{sub 3} slurry was prepared using a commercial powder of particle size 0.5–1.5 μm and the pebble pre-form was prepared by dropping the slurry into liquid nitrogen through a syringe needle. The droplets were rapidly frozen, changing their morphology to spherical pebbles. The frozen pebbles were dried at −10 °C in vacuum. To make crack-free pebbles, some glycerin was employed in the slurry, and long drying time and a low vacuum condition were applied in the freeze drying process. In the process, the solid content in the slurry influenced the spheroidicity of the pebble green body. The dried pebbles were sintered at 1200 °C in an air atmosphere. The sintered pebbles showed almost 40% shrinkage. The sintered pebbles revealed a porous microstructure with a uniform pore distribution and the sintered pebbles were crushed under an average load of 50 N in a compressive strength test. In the present study, a freeze drying process for fabrication of spherical Li{sub 2}TiO{sub 3} pebbles is introduced. The processing parameters, such as solid content in the slurry and the conditions of freeze drying and sintering, are also examined.

  13. Study on dewatering of chemical sludge by freeze-thaw process

    International Nuclear Information System (INIS)

    Xu Shikun; Liu Pin

    1993-01-01

    The treatment of radioactive sludge that is produced from treating radioactive waste water contains radioactively is different from that of non-radioactive sludge. The methods of immersing freeze and simulated two-step freeze have been studied for the elementary properties of simulated low-level radioactive sledge, the effect of freezing temperature, freeze time, and settling time on volume-reduction factor. Some parameters for design of freeze-thaw device are provided

  14. Immobilization of sericin molecules via amorphous carbon plasma modified-polystyrene dish for serum-free culture

    International Nuclear Information System (INIS)

    Tunma, Somruthai; Song, Doo-Hoon; Kim, Si-Eun; Kim, Kyoung-Nam; Han, Jeon-Geon; Boonyawan, Dheerawan

    2013-01-01

    In this study, we focused on sericin hydrolysates, originating from silkworm used in serum-free human bone marrow-derived mesenchymal stem cells (hBM-MSCs) culture. We reported the effect of a covalent linkage between a bioactive protein molecule and polystyrene dish surface via a carbon intermediate layer which can slow down the release rate of protein compounds into the phosphate buffer saline (PBS) solution. Films of amorphous carbon (a-C) and functionalized-carbon were deposited on PS culture dish surfaces by using a DC magnetron sputtering system and RF PECVD system. We found that a-C based-films can increase the hydrophilicity and biocompatibility of polystyrene (PS) dishes, especially a-C films and a-C:N 2 films showed good attachment of hBM-MSCs at 24 h. However, in the case of silica surface (a-C:SiO x films), the cells showed a ragged and unattached boundary resulting from the presence of surface silanol groups. For the UV–vis absorbance, all carbon modified-PS dishes showed a lower release rate of sericin molecules into PBS solution than PS control. This revealed that the functionalized carbon could be enhanced by specific binding properties with given molecules. The carbon-coated PS dishes grafting with sericin protein were used in a serum-free condition. We also found that hBM-MSCs have higher percentage of proliferated cells at day 7 for the modified dishes with carbon films and coated with sericin than the PS control coated with sericin. The physical film properties were measured by atomic force microscopy (AFM), scanning electron microscope (SEM) and contact angle measurement. The presence of -NH 2 groups of sericin compounds on the PS dish was revealed by Fourier transform infrared spectroscopy (FTIR). The stability of covalent bonds of sericin molecules after washing out ungrafted sericin was confirmed by X-ray photoelectron spectroscopy (XPS).

  15. Immobilization of sericin molecules via amorphous carbon plasma modified-polystyrene dish for serum-free culture

    Science.gov (United States)

    Tunma, Somruthai; Song, Doo-Hoon; Kim, Si-Eun; Kim, Kyoung-Nam; Han, Jeon-Geon; Boonyawan, Dheerawan

    2013-10-01

    In this study, we focused on sericin hydrolysates, originating from silkworm used in serum-free human bone marrow-derived mesenchymal stem cells (hBM-MSCs) culture. We reported the effect of a covalent linkage between a bioactive protein molecule and polystyrene dish surface via a carbon intermediate layer which can slow down the release rate of protein compounds into the phosphate buffer saline (PBS) solution. Films of amorphous carbon (a-C) and functionalized-carbon were deposited on PS culture dish surfaces by using a DC magnetron sputtering system and RF PECVD system. We found that a-C based-films can increase the hydrophilicity and biocompatibility of polystyrene (PS) dishes, especially a-C films and a-C:N2 films showed good attachment of hBM-MSCs at 24 h. However, in the case of silica surface (a-C:SiOx films), the cells showed a ragged and unattached boundary resulting from the presence of surface silanol groups. For the UV-vis absorbance, all carbon modified-PS dishes showed a lower release rate of sericin molecules into PBS solution than PS control. This revealed that the functionalized carbon could be enhanced by specific binding properties with given molecules. The carbon-coated PS dishes grafting with sericin protein were used in a serum-free condition. We also found that hBM-MSCs have higher percentage of proliferated cells at day 7 for the modified dishes with carbon films and coated with sericin than the PS control coated with sericin. The physical film properties were measured by atomic force microscopy (AFM), scanning electron microscope (SEM) and contact angle measurement. The presence of sbnd NH2 groups of sericin compounds on the PS dish was revealed by Fourier transform infrared spectroscopy (FTIR). The stability of covalent bonds of sericin molecules after washing out ungrafted sericin was confirmed by X-ray photoelectron spectroscopy (XPS).

  16. Hot big bang or slow freeze?

    Science.gov (United States)

    Wetterich, C.

    2014-09-01

    We confront the big bang for the beginning of the universe with an equivalent picture of a slow freeze - a very cold and slowly evolving universe. In the freeze picture the masses of elementary particles increase and the gravitational constant decreases with cosmic time, while the Newtonian attraction remains unchanged. The freeze and big bang pictures both describe the same observations or physical reality. We present a simple ;crossover model; without a big bang singularity. In the infinite past space-time is flat. Our model is compatible with present observations, describing the generation of primordial density fluctuations during inflation as well as the present transition to a dark energy-dominated universe.

  17. Using CR1aa versus KSOM as the culture medium for in vitro embryo production of cattle

    Directory of Open Access Journals (Sweden)

    Endang Triwulaninngsih

    2002-03-01

    Full Text Available This research has been conducted at the laboratory of in vitro fertilization in the Department of Animal Science University of Wisconsin, USA. These embryos can be used for improving genetic value of Indonesian cattle. Oocytes were matured in TCM- 199 medium (in 5% CO2 incubator and at 390C enriched with follicle stimulating hormone (FSH 10 μl/ml, oestradiol 17 β 1μl/ml and 10% Fetal Calf Serum (FCS. The oocytes were fertilized in vitro with motile sperm and incubation between sperm and oocytes in fertilization medium Tyroide Albumin Lactate Pyruvate (TALP for 20 hours. All zygotes were cultured in CR1aa (n=1549 medium versus modification of protein-free pottasium simplex optimized medium (KSOM (n=675 up to blastocyst stage and were fed FCS 5 μl/50 μl medium on day 6, as treatment A and B respectively. Data were analyzed by completely randomized design with SAS program. Percentages of cleavage, morula, blastocyst, expanded blastocyst, unfertilized and degenerated ova in this study were 91.4% vs 75.6 %; 75.6% vs 58.9%; 61.5% vs 38.5%; 31.2% vs 5.1%, 8.6% vs 24.4%, 15.7% vs 8% which were significantly different (P<0.01 for treatment CR1aa and KSOM respectively. Based on this study, CR1aa medium is better culture medium than KSOM for efficient in vitro production (IVP of bovine embryos.

  18. Freeze dried bone allografts in dental and maxillofacial reconstructive surgery - experience in Malaysia

    International Nuclear Information System (INIS)

    Abd Rani Samsudin; Meor Zaidi Meor Kamal

    1999-01-01

    The utilisation of vascularised and free bone autografts remain the goal standard in maxillofacial reconstructive surgery in Malaysia, but the use of freeze dried bone allograft is still widely practiced in many centres with variable results. This study evaluate the effectiveness and clinical efficacy of using radiation sterilised freeze dried bone allografts in oral and maxillofacial reconstructive surgery. The bone grafts were prepared at the Malaysian National Tissue Bank. Seventy eight patients who had undergone oral and Maxillofacial surgical procedures with reconstruction using bone allografts were included in this study. 50 patients were male and 28 patients were female and their age ranged from 14 to 75 years. Forty two patients underwent enucleation of benign cystic lesions in the jaws, 15 patients underwent repair of orbital floor fractures, 6 patients of jaw fractures with partial loss of bone while 8 patients underwent augmentation of depressed cheek bone. Another 4 patients had partial resection of the mandible because of cancer and 3 patients had facial osteotomies. A follow up period of 12 months up to 4 years was carried out. The patients were assessed both clinically and radiologically throughout their follow up visits. Clinical assessment showed no evidence of rejection of the implanted freeze dried allografts. Bone allografts implanted as inlay grafts demonstrated a better clinical performance than onlay grafts and the poorest results were obtained following bridging bony defects in the jaws. Radiation sterilised freeze dried bone allografts produced at the Malaysian National Tissue Bank are bio-compatible, functional, and provide predictable results when applied to selected areas of the facial skeleton

  19. Recent developments in smart freezing technology applied to fresh foods.

    Science.gov (United States)

    Xu, Ji-Cheng; Zhang, Min; Mujumdar, Arun S; Adhikari, Benu

    2017-09-02

    Due to the increased awareness of consumers in sensorial and nutritional quality of frozen foods, the freezing technology has to seek new and innovative technologies for better retaining the fresh like quality of foods. In this article, we review the recent developments in smart freezing technology applied to fresh foods. The application of these intelligent technologies and the associated underpinning concepts have greatly improved the quality of frozen foods and the freezing efficiency. These technologies are able to automatically collect the information in-line during freezing and help control the freezing process better. Smart freezing technology includes new and intelligent technologies and concepts applied to the pretreatment of the frozen product, freezing processes, cold chain logistics as well as warehouse management. These technologies enable real-time monitoring of quality during the freezing process and help improve product quality and freezing efficiency. We also provide a brief overview of several sensing technologies used to achieve automatic control of individual steps of freezing process. These sensing technologies include computer vision, electronic nose, electronic tongue, digital simulation, confocal laser, near infrared spectroscopy, nuclear magnetic resonance technology and ultrasound. Understanding of the mechanism of these new technologies will be helpful for applying them to improve the quality of frozen foods.

  20. Correction factors for the NMi free-air ionization chamber for medium-energy x-rays calculated with the Monte Carlo method

    International Nuclear Information System (INIS)

    Grimbergen, T.W.M.; Dijk, E. van; Vries, W. de

    1998-01-01

    A new method is described for the determination of x-ray quality dependent correction factors for free-air ionization chambers. The method is based on weighting correction factors for mono-energetic photons, which are calculated using the Monte Carlo method, with measured air kerma spectra. With this method, correction factors for electron loss, scatter inside the chamber and transmission through the diaphragm and front wall have been calculated for the NMi free-air chamber for medium-energy x-rays for a wide range of x-ray qualities in use at NMi. The newly obtained correction factors were compared with the values in use at present, which are based on interpolation of experimental data for a specific set of x-ray qualities. For x-ray qualities which are similar to this specific set, the agreement between the correction factors determined with the new method and those based on the experimental data is better than 0.1%, except for heavily filtered x-rays generated at 250 kV. For x-ray qualities dissimilar to the specific set, differences up to 0.4% exist, which can be explained by uncertainties in the interpolation procedure of the experimental data. Since the new method does not depend on experimental data for a specific set of x-ray qualities, the new method allows for a more flexible use of the free-air chamber as a primary standard for air kerma for any x-ray quality in the medium-energy x-ray range. (author)

  1. Label-free LC-MSe in tissue and serum reveals protein networks underlying differences between benign and malignant serous ovarian tumors.

    Directory of Open Access Journals (Sweden)

    Wouter Wegdam

    Full Text Available PURPOSE: To identify proteins and (molecular/biological pathways associated with differences between benign and malignant epithelial ovarian tumors. EXPERIMENTAL PROCEDURES: Serum of six patients with a serous adenocarcinoma of the ovary was collected before treatment, with a control group consisting of six matched patients with a serous cystadenoma. In addition to the serum, homogeneous regions of cells exhibiting uniform histology were isolated from benign and cancerous tissue by laser microdissection. We subsequently employed label-free liquid chromatography tandem mass spectrometry (LC-MSe to identify proteins in these serum and tissues samples. Analyses of differential expression between samples were performed using Bioconductor packages and in-house scripts in the statistical software package R. Hierarchical clustering and pathway enrichment analyses were performed, as well as network enrichment and interactome analysis using MetaCore. RESULTS: In total, we identified 20 and 71 proteins that were significantly differentially expressed between benign and malignant serum and tissue samples, respectively. The differentially expressed protein sets in serum and tissue largely differed with only 2 proteins in common. MetaCore network analysis, however inferred GCR-alpha and Sp1 as common transcriptional regulators. Interactome analysis highlighted 14-3-3 zeta/delta, 14-3-3 beta/alpha, Alpha-actinin 4, HSP60, and PCBP1 as critical proteins in the tumor proteome signature based on their relative overconnectivity. The data have been deposited to the ProteomeXchange with identifier PXD001084. DISCUSSION: Our analysis identified proteins with both novel and previously known associations to ovarian cancer biology. Despite the small overlap between differentially expressed protein sets in serum and tissue, APOA1 and Serotransferrin were significantly lower expressed in both serum and cancer tissue samples, suggesting a tissue-derived effect in serum

  2. Nanoparticles of Conjugated Methotrexate-Human Serum Albumin: Preparation and Cytotoxicity Evaluations

    Directory of Open Access Journals (Sweden)

    Azade Taheri

    2011-01-01

    Full Text Available Methotrexate-human serum albumin conjugates were developed by a simple carbodiimide reaction. Methotrexate-human serum albumin conjugates were then crosslinked with 1-ethyl-3-(3-dimethylaminopropyl carbodiimide HCl (EDC to form nanoparticles. The size of nanoparticles determined by laser light scattering and TEM was between 90–150 nm. Nanoparticles were very stable at physiologic conditions (PBS pH 7.4, 37∘C and after incubation with serum. The effect of amount of EDC used for crosslinking on the particle size and free amino groups of nanoparticles was examined. The amount of crosslinker showed no significant effect on the size of nanoparticles but free amino groups of nanoparticles were decreased by increasing the crosslinker. The physicochemical interactions between methotrexate and human serum albumin were investigated by differential scanning calorimetry (DSC. Nanoparticles were more cytotoxic on T47D cells compared to free methotrexate. Moreover, methotrexate-human serum albumin nanoparticles decreased the IC50 value of methotrexate on T47D cells in comparison with free methotrexate.

  3. Nanoparticles of Conjugated Methotrexate-Human Serum Albumin: Preparation and Cytotoxicity Evaluations

    International Nuclear Information System (INIS)

    Taheri, A.; Atyabi, F.; Nouri, F.S.; Ahadi, F.; Derakhshan, M.A.; Dinarvand, R.; Atyabi, F.; Ghahremani, M.H.; Ostad, S.N.; Dinarvand, R.; Amini, M.; Ghahremani, M.H.; Ostad, S.N.; Mansoori, P.

    2011-01-01

    Methotrexate-human serum albumin conjugates were developed by a simple carbodiimide reaction. Methotrexate-human serum albumin conjugates were then crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide HCl (EDC) to form nanoparticles. The size of nanoparticles determined by laser light scattering and TEM was between 90 150 nm. Nanoparticles were very stable at physiologic conditions (PBS pH 7.4, 37 degree C) and after incubation with serum. The effect of amount of EDC used for crosslinking on the particle size and free amino groups of nanoparticles was examined. The amount of cross linker showed no significant effect on the size of nanoparticles but free amino groups of nanoparticles were decreased by increasing the cross linker. The physicochemical interactions between methotrexate and human serum albumin were investigated by differential scanning calorimetry (DSC). Nanoparticles were more cytotoxic on T 47 D cells compared to free methotrexate. Moreover, methotrexate-human serum albumin nanoparticles decreased the C50 value of methotrexate on T 47 D cells in comparison with free methotrexate.

  4. Freeze-all cycle for all normal responders?

    Science.gov (United States)

    Roque, Matheus; Valle, Marcello; Guimarães, Fernando; Sampaio, Marcos; Geber, Selmo

    2017-02-01

    The purpose of this study is to evaluate the freeze-all strategy in subgroups of normal responders, to assess whether this strategy is beneficial regardless of ovarian response, and to evaluate the possibility of implementing an individualized embryo transfer (iET) based on ovarian response. This was an observational, cohort study performed in a private IVF center. A total of 938 IVF cycles were included in this study. The patients were submitted to controlled ovarian stimulation (COS) with a gonadotropin-releasing hormone (GnRH) antagonist protocol and a cleavage-stage day 3 embryo transfer. We performed a comparison of outcomes between the fresh embryo transfer (n = 523) and the freeze-all cycles (n = 415). The analysis was performed in two subgroups of patients based on the number of retrieved oocytes: Group 1 (4-9 oocytes) and Group 2 (10-15 oocytes). In Group 1 (4-9 retrieved oocytes), the implantation rates (IR) were 17.9 and 20.5% (P = 0.259) in the fresh and freeze-all group, respectively; the ongoing pregnancy rates (OPR) were 31 and 33% (P = 0.577) in the fresh and freeze-all group, respectively. In Group 2 (10-15 oocytes), the IR were 22.1 and 30.1% (P = 0.028) and the OPR were 34 and 47% (P = 0.021) in the fresh and freeze-all groups, respectively. Although the freeze-all policy may be related to better in vitro fertilization (IVF) outcomes in normal responders, these potential advantages decrease with worsening ovarian response. Patients with poorer ovarian response do not benefit from the freeze-all strategy.

  5. Rational design of an influenza subunit vaccine powder with sugar glass technology : preventing conformational changes of haemagglutinin during freezing and freeze-drying

    NARCIS (Netherlands)

    Amorij, J-P; Meulenaar, J; Hinrichs, W L J; Stegmann, T; Huckriede, A; Coenen, F; Frijlink, H W

    2007-01-01

    The development of a stable influenza subunit vaccine in the dry state was investigated. The influence of various carbohydrates, buffer types and freezing rates on the integrity of haemagglutinin after freeze-thawing or freeze-drying was investigated with a range of analytical and immunological

  6. Efficient odd straight medium chain free fatty acid production by metabolically engineered Escherichia coli.

    Science.gov (United States)

    Wu, Hui; San, Ka-Yiu

    2014-11-01

    Free fatty acids (FFAs) can be used as precursors for the production of biofuels or chemicals. Different composition of FFAs will be useful for further modification of the biofuel/biochemical quality. Microbial biosynthesis of even chain FFAs can be achieved by introducing an acyl-acyl carrier protein thioesterase gene into E. coli. In this study, odd straight medium chain FFAs production was investigated by using metabolic engineered E. coli carrying acyl-ACP thioesterase (TE, Ricinus communis), propionyl-CoA synthase (Salmonella enterica), and β-ketoacyl-acyl carrier protein synthase III (four different sources) with supplement of extracellular propionate. By using these metabolically engineered E. coli, significant quantity of C13 and C15 odd straight-chain FFAs could be produced from glucose and propionate. The highest concentration of total odd straight chain FFAs attained was 1205 mg/L by the strain HWK201 (pXZ18, pBHE2), and 85% of the odd straight chain FFAs was C15. However, the highest percentage of odd straight chain FFAs was achieved by the strain HWK201 (pXZ18, pBHE3) of 83.2% at 48 h. This strategy was also applied successfully in strains carrying different TE, such as the medium length acyl-ACP thioesterase gene from Umbellularia californica. C11 and C13 became the major odd straight-chain FFAs. © 2014 Wiley Periodicals, Inc.

  7. Medical and social egg freezing

    DEFF Research Database (Denmark)

    Lallemant, Camille; Vassard, Ditte; Andersen, Anders Nyboe

    2016-01-01

    INTRODUCTION: Until recently, limited options for preserving fertility in order to delay childbearing were available. Although egg freezing and successful thawing is now possible, it remains unclear to what extent women are aware of the availability of this technique, their attitudes towards its...... use, or the circumstances under which this technique may be considered. MATERIAL AND METHODS: An online cross-sectional survey was designed to investigate knowledge and attitudes of women in Denmark and the UK on egg freezing and their potential intentions regarding the procedure. RESULTS: Data...... was collected from September 2012 to September 2013 and the responses of 973 women were analyzed. In total, 83% of women reported having heard of egg freezing, and nearly all considered it acceptable for medical indications, whilst 89% considered it acceptable for social reasons. Overall, 19% expressed active...

  8. Response of seasonal soil freeze depth to climate change across China

    Science.gov (United States)

    Peng, Xiaoqing; Zhang, Tingjun; Frauenfeld, Oliver W.; Wang, Kang; Cao, Bin; Zhong, Xinyue; Su, Hang; Mu, Cuicui

    2017-05-01

    The response of seasonal soil freeze depth to climate change has repercussions for the surface energy and water balance, ecosystems, the carbon cycle, and soil nutrient exchange. Despite its importance, the response of soil freeze depth to climate change is largely unknown. This study employs the Stefan solution and observations from 845 meteorological stations to investigate the response of variations in soil freeze depth to climate change across China. Observations include daily air temperatures, daily soil temperatures at various depths, mean monthly gridded air temperatures, and the normalized difference vegetation index. Results show that soil freeze depth decreased significantly at a rate of -0.18 ± 0.03 cm yr-1, resulting in a net decrease of 8.05 ± 1.5 cm over 1967-2012 across China. On the regional scale, soil freeze depth decreases varied between 0.0 and 0.4 cm yr-1 in most parts of China during 1950-2009. By investigating potential climatic and environmental driving factors of soil freeze depth variability, we find that mean annual air temperature and ground surface temperature, air thawing index, ground surface thawing index, and vegetation growth are all negatively associated with soil freeze depth. Changes in snow depth are not correlated with soil freeze depth. Air and ground surface freezing indices are positively correlated with soil freeze depth. Comparing these potential driving factors of soil freeze depth, we find that freezing index and vegetation growth are more strongly correlated with soil freeze depth, while snow depth is not significant. We conclude that air temperature increases are responsible for the decrease in seasonal freeze depth. These results are important for understanding the soil freeze-thaw dynamics and the impacts of soil freeze depth on ecosystem and hydrological process.

  9. Effects of freezing conditions on quality changes in blueberries.

    Science.gov (United States)

    Cao, Xuehui; Zhang, Fangfang; Zhao, Dongyu; Zhu, Danshi; Li, Jianrong

    2018-03-12

    Freezing preservation is one of the most effective methods used to maintain the flavour and nutritional value of fruit. This research studied the effects of different freezing conditions, -20 °C, -40 °C, -80 °C, and immersion in liquid nitrogen, on quality changes of freeze-thawed blueberries. The water distribution estimates of blueberries were measured based on low-field nuclear magnetic resonance (LF-NMR) analysis. The pectin content, drip loss, and fruit texture were also detected to evaluate quality changes in samples. The freezing curves of blueberry showed super-cooling points at -20 °C and - 40 °C, whereas super-cooling points were not observed at -80 °C or in liquid nitrogen. After freeze-thaw treatment, the relaxation time of the cell wall water (T 21 ), cytoplasm water and extracellular space (T 22 ), and vacuole water (T 23 ) were significantly shortened compared to fresh samples, which suggested a lower liquidity. Although the freezing speed for samples immersed in liquid nitrogen was faster than other treatments, samples treated at -80 °C showed better quality regarding vacuole water holding, drip loss, and original pectin content retention. This study contributed to understanding how freezing temperature affects the qualities of blueberries. The super-fast freezing rate might injure fruit, and an appropriate freezing rate could better preserve blueberries. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  10. The freezing and supercooling of garlic (Allium sativum L.)

    Energy Technology Data Exchange (ETDEWEB)

    James, Christian; Seignemartin, Violaine; James, Stephen J. [Food Refrigeration and Process Engineering Research Centre (FRPERC), University of Bristol, Churchill Building, Langford, Bristol BS40 5DU (United Kingdom)

    2009-03-15

    This work shows that peeled garlic cloves demonstrate significant supercooling during freezing under standard conditions and can be stored at temperatures well below their freezing point (-2.7 C) without freezing. The nucleation point or 'metastable limit temperature' (the point at which ice crystal nucleation is initiated) of peeled garlic cloves was found to be between -7.7 and -14.6 C. Peeled garlic cloves were stored under static air conditions at temperatures between -6 and -9 C for up to 69 h without freezing, and unpeeled whole garlic bulbs and cloves were stored for 1 week at -6 C without freezing. (author)

  11. Effect of freezing conditions on β-Tricalcium Phosphate /Camphene scaffold with micro sized particles fabricated by freeze casting.

    Science.gov (United States)

    Singh, Gurdev; Soundarapandian, S

    2018-03-01

    The long standing need of the implant manufacturing industries is to fabricate multi-matrix, customized porous scaffold as cost-effectively. In recent years, freeze casting has shown greater opportunity in the fabrication of porous scaffolds (tricalcium phosphate, hydroxyapatite, bioglass, alumina, etc.) such as at ease and good control over pore size, porosity, a range of materials and economic feasibility. In particular, tricalcium phosphate (TCP) has proved as it possesses good biocompatible (osteoinduction, osteoconduction, etc.) and biodegradability hence beta-tricalcium phosphate (β-TCP, particle size of 10µm) was used as base material and camphene was used as a freezing vehicle in this study. Both freezing conditions such as constant freezing temperature (CFT) and constant freezing rate (CFR) were used for six different conditional samples (CFT: 30, 35 and 40vol% solid loading; similarly CFR: 30, 35 and 40vol% solid loading) to study and understand the effect of various properties (pore size, porosity and compressive strength) of the freeze-cast porous scaffold. It was observed that the average size of the pore was varying linearly as from lower to higher when the solid loading was varying higher to lower. With the help of scanning electron micrographs (SEM), it was observed that the average size of pore during CFR (9.7/ 6.5/ 4.9µm) was comparatively higher than the process of CFT (6.0/ 4.8/ 2.6µm) with respect to the same solid loading (30/ 35/ 40vol%) conditions. From the Gas pycnometer analysis, it was found that the porosity in both freezing conditions (CFT, CFR) were almost near values such as 32.8% and 28.5%. Further to be observed that with the increase in solid loading, the total porosity value has decreased due to the reduction in the concentration of the freezing vehicle. Hence, the freezing vehicle was found as responsible for the formation of appropriate size and orientation of pores during freeze casting. The compressive strength (CS

  12. Technological Development of Brewing in Domestic Refrigerator Using Freeze-Dried Raw Materials

    Directory of Open Access Journals (Sweden)

    Angelika-Ioanna Gialleli

    2017-01-01

    Full Text Available Development of a novel directly marketable beer brewed at low temperature in a domestic refrigerator combined with yeast immobilization technology is presented in this study. Separately, freeze-dried wort and immobilized cells of the cryotolerant yeast strain Saccharomyces cerevisiae AXAZ-1 on tubular cellulose were used in low-temperature fermentation (2, 5 and 7 °C. The positive eff ect of tubular cellulose during low-temperature brewing was examined, revealing that freeze-dried immobilized yeast cells on tubular cellulose signifi cantly reduced the fermentation rates in contrast to freeze-dried free cells, although they are recommended for home-made beer production. Immobilization also enhanced the yeast resistance at low-temperature fermentation, reducing the minimum brewing temperature value from 5 to 2 °C. In the case of high-quality beer production, the eff ect of temperature and initial sugar concentration on the fermentation kinetics were assessed. Sensory enrichment of the produced beer was confi rmed by the analysis of the fi nal products, revealing a low diacetyl concentration, together with improved polyphenol content, aroma profi le and clarity. The proposed process for beer production in a domestic refrigerator can easily be commercialized and applied by dissolving the content of two separate packages in tap water; one package containing dried wort and the other dried immobilized cells on tubular cellulose suspended in tap water.

  13. Technological Development of Brewing in Domestic Refrigerator Using Freeze-Dried Raw Materials.

    Science.gov (United States)

    Gialleli, Angelika-Ioanna; Ganatsios, Vassilios; Terpou, Antonia; Kanellaki, Maria; Bekatorou, Argyro; Koutinas, Athanasios A; Dimitrellou, Dimitra

    2017-09-01

    Development of a novel directly marketable beer brewed at low temperature in a domestic refrigerator combined with yeast immobilization technology is presented in this study. Separately, freeze-dried wort and immobilized cells of the cryotolerant yeast strain Saccharomyces cerevisiae AXAZ-1 on tubular cellulose were used in low-temperature fermentation (2, 5 and 7 °C). The positive effect of tubular cellulose during low-temperature brewing was examined, revealing that freeze-dried immobilized yeast cells on tubular cellulose significantly reduced the fermentation rates in contrast to freeze-dried free cells, although they are recommended for home-made beer production. Immobilization also enhanced the yeast resistance at low-temperature fermentation, reducing the minimum brewing temperature value from 5 to 2 °C. In the case of high-quality beer production, the effect of temperature and initial sugar concentration on the fermentation kinetics were assessed. Sensory enrichment of the produced beer was confirmed by the analysis of the final products, revealing a low diacetyl concentration, together with improved polyphenol content, aroma profile and clarity. The proposed process for beer production in a domestic refrigerator can easily be commercialized and applied by dissolving the content of two separate packages in tap water; one package containing dried wort and the other dried immobilized cells on tubular cellulose suspended in tap water.

  14. Simulation of Changes in the Near-Surface Soil Freeze/Thaw Cycle Using CLM4.5 With Four Atmospheric Forcing Data Sets

    Science.gov (United States)

    Guo, Donglin; Wang, Aihui; Li, Duo; Hua, Wei

    2018-03-01

    Change in the near-surface soil freeze/thaw cycle is critical for assessments of hydrological activity, ecosystems, and climate change. Previous studies investigated the near-surface soil freeze/thaw cycle change mostly based on in situ observations and satellite monitoring. Here numerical simulation method is tested to estimate the long-term change in the near-surface soil freeze/thaw cycle in response to recent climate warming for its application to predictions. Four simulations are performed at 0.5° × 0.5° resolution from 1979 to 2009 using the Community Land Model version 4.5, each driven by one of the four atmospheric forcing data sets (i.e., one default Climate Research Unit-National Centers for Environmental Prediction [CRUNCEP] and three newly developed Modern Era Retrospective-Analysis for Research and Applications, Climate Forecast System Reanalysis, and European Centre for Medium-Range Weather Forecasts Reanalysis Interim). The observations from 299 weather stations in both Russia and China are employed to validate the simulated results. The results show that all simulations reasonably reproduce the observed variations in the ground temperature, the freeze start and end dates, and the freeze duration (the correlation coefficients range from 0.47 to 0.99, and the Nash-Sutcliffe efficiencies range from 0.19 to 0.98). Part of the simulations also exactly simulate the trends of the ground temperature, the freeze start and end dates, and the freeze duration. Of the four simulations, the results from the simulation using the CRUNCEP data set show the best overall agreement with the in situ observations, indicating that the CRUNCEP data set could be preferentially considered as the basic atmospheric forcing data set for future prediction. The simulated area-averaged annual freeze duration shortened by 8.03 days on average from 1979 to 2009, with an uncertainty (one standard deviation) of 0.67 days caused by the different atmospheric forcing data sets. These

  15. Evaluation of the Efficiency of Two Different Freezing Media and Two Different Protocols to Preserve Human Spermatozoa from Cryoinjury

    Directory of Open Access Journals (Sweden)

    Gemma Fabozzi

    2016-01-01

    Full Text Available It is universally recognized that cryopreservation impairs sperm quality. In order to improve postthawing sperm survival and motility, media of different composition and different protocols have been proposed. However, no clear evidence is available to understand which are the most efficient protocol and medium for sperm cryopreservation. The present study evaluates the efficiency of two different cryopreservation protocols and two common freezing media (FM containing different cryoprotectants (CPs, TEST Yolk Buffer (TYB and Sperm Freeze (SF, to preserve human sperm quality. Our data suggest that TYB is better than SF both in terms of postthaw viability and in terms of progressive motility, while the direct addition of FM to the sperm sample resulted in the most efficient protocol in terms of postthaw viability but not in terms of progressive motility.

  16. The choice of a suitable oligosaccharide to prevent aggregation of PEGylated nanoparticles during freeze thawing and freeze drying

    NARCIS (Netherlands)

    Hinrichs, Wouter; Manceñido, F A; Sanders, N N; Braeckmans, K; De Smedt, S C; Demeester, J; Frijlink, H W

    2006-01-01

    In a previous Study we have shown that the oligosaccharide inulin can prevent aggregation of poly(ethylene glycol) (PEG) coated plasmid DNA/cationic liposome complexes ('' PEGylated lipoplexes '') during freeze thawing and freeze drying [Hinrichs et al., 2005. J. Control. Release 103, 465]. By

  17. Association of PSA, free-PSA and testosteron levels in serum of patients with benign prostate hyperplasia (BPH) and prostate cancer

    International Nuclear Information System (INIS)

    Wiwin Mailana; Kristina Dwi P; Sri Insani WW; Puji Widayati

    2015-01-01

    Prostate cancer screening can be done by measuring the concentration levels of PSA, free-PSA and testosterone in serum that examined with radioimmunoassay (RIA). A total of 30 patients of 45-81 years old had enrolled in this study and were taken their venous blood. The aim of research is to know the relationship between PSA and testosterone free-PSA with BPH and prostate cancer. Results showed that there was no correlation between age with BPH and prostate cancer (p = 0.06), but there is a relationship between PSA with BPH and prostate cancer (p = 0.002), the relationship between free-PSA with BPH and prostate cancer (p = 0.001). No correlation was found between PSA ratio with BPH and prostate cancer as well as the absence of a relationship between testosterone with BPH and prostate cancer (p = 0.924). (author)

  18. Antimicrobial medium- and long-chain free fatty acids prevent PrfA-dependent activation of virulence genes in Listeria monocytogenes

    DEFF Research Database (Denmark)

    Sternkopf Lillebæk, Eva Maria; Lambert Nielsen, Stine; Scheel Thomasen, Rikke

    2017-01-01

    of virulence factors required for bacterial entry, intracellular replication and cell-to-cell spread. PrfA-dependent activation of virulence genes occurs primarily in the blood and during intracellular infection. In contrast, PrfA does not play a significant role in regulation of virulence gene expression...... antimicrobial free fatty acids act to downregulate transcription of PrfA-activated virulence genes. Interestingly, the inhibitory effect is also evident in cells encoding a constitutively active variant of PrfA. Collectively, our data suggest that antimicrobial medium- and long-chain free fatty acids may act...... as signals to prevent PrfA-mediated activation of virulence genes in environments where PrfA activation is not required, such as in food and the gastrointestinal tract....

  19. Faktor Influencing the Vacuum Freezing Rate of Liquid Food

    OpenAIRE

    Tambunan, Armansyah H

    2000-01-01

    Many,freezing methods, mechanicul as well as cryogenic, have been in wide application in food industries. Each method has its own advantage, but in regard with the food quality, freezing rule can be accomplished by the method is one of the tnost important factors. Nowadays, many researchers are conducting experiment in order to enhance thefi.eezing rate. This paper deals with the advantage of vacuum freezing method in enhancing the freezing rate and its applicability for liquidfood.Experinren...

  20. Increased spring freezing vulnerability for alpine shrubs under early snowmelt.

    Science.gov (United States)

    Wheeler, J A; Hoch, G; Cortés, A J; Sedlacek, J; Wipf, S; Rixen, C

    2014-05-01

    Alpine dwarf shrub communities are phenologically linked with snowmelt timing, so early spring exposure may increase risk of freezing damage during early development, and consequently reduce seasonal growth. We examined whether environmental factors (duration of snow cover, elevation) influenced size and the vulnerability of shrubs to spring freezing along elevational gradients and snow microhabitats by modelling the past frequency of spring freezing events. We sampled biomass and measured the size of Salix herbacea, Vaccinium myrtillus, Vaccinium uliginosum and Loiseleuria procumbens in late spring. Leaves were exposed to freezing temperatures to determine the temperature at which 50% of specimens are killed for each species and sampling site. By linking site snowmelt and temperatures to long-term climate measurements, we extrapolated the frequency of spring freezing events at each elevation, snow microhabitat and per species over 37 years. Snowmelt timing was significantly driven by microhabitat effects, but was independent of elevation. Shrub growth was neither enhanced nor reduced by earlier snowmelt, but decreased with elevation. Freezing resistance was strongly species dependent, and did not differ along the elevation or snowmelt gradient. Microclimate extrapolation suggested that potentially lethal freezing events (in May and June) occurred for three of the four species examined. Freezing events never occurred on late snow beds, and increased in frequency with earlier snowmelt and higher elevation. Extrapolated freezing events showed a slight, non-significant increase over the 37-year record. We suggest that earlier snowmelt does not enhance growth in four dominant alpine shrubs, but increases the risk of lethal spring freezing exposure for less freezing-resistant species.

  1. Hot big bang or slow freeze?

    Energy Technology Data Exchange (ETDEWEB)

    Wetterich, C.

    2014-09-07

    We confront the big bang for the beginning of the universe with an equivalent picture of a slow freeze — a very cold and slowly evolving universe. In the freeze picture the masses of elementary particles increase and the gravitational constant decreases with cosmic time, while the Newtonian attraction remains unchanged. The freeze and big bang pictures both describe the same observations or physical reality. We present a simple “crossover model” without a big bang singularity. In the infinite past space–time is flat. Our model is compatible with present observations, describing the generation of primordial density fluctuations during inflation as well as the present transition to a dark energy-dominated universe.

  2. Hot big bang or slow freeze?

    International Nuclear Information System (INIS)

    Wetterich, C.

    2014-01-01

    We confront the big bang for the beginning of the universe with an equivalent picture of a slow freeze — a very cold and slowly evolving universe. In the freeze picture the masses of elementary particles increase and the gravitational constant decreases with cosmic time, while the Newtonian attraction remains unchanged. The freeze and big bang pictures both describe the same observations or physical reality. We present a simple “crossover model” without a big bang singularity. In the infinite past space–time is flat. Our model is compatible with present observations, describing the generation of primordial density fluctuations during inflation as well as the present transition to a dark energy-dominated universe

  3. Hot big bang or slow freeze?

    Directory of Open Access Journals (Sweden)

    C. Wetterich

    2014-09-01

    Full Text Available We confront the big bang for the beginning of the universe with an equivalent picture of a slow freeze — a very cold and slowly evolving universe. In the freeze picture the masses of elementary particles increase and the gravitational constant decreases with cosmic time, while the Newtonian attraction remains unchanged. The freeze and big bang pictures both describe the same observations or physical reality. We present a simple “crossover model” without a big bang singularity. In the infinite past space–time is flat. Our model is compatible with present observations, describing the generation of primordial density fluctuations during inflation as well as the present transition to a dark energy-dominated universe.

  4. FREEZE DRYING PROCESS: A REVIEW

    OpenAIRE

    Soham Shukla

    2011-01-01

    Among the various methods of drying, this article has mentioned only one most important method, “Freeze drying”. This method is mainly used for the drying of thermo labile materials. This method works on the principle of sublimation. This method is divided into 3 steps for its better understanding; these are Freezing, Primary drying, and secondary drying. There are many advantages and disadvantages of this method, but still this is the most useful drying method nowadays.

  5. Effects of protectant and rehydration conditions on the survival rate and malolactic fermentation efficiency of freeze-dried Lactobacillus plantarum JH287.

    Science.gov (United States)

    Lee, Sae-Byuk; Kim, Dong-Hwan; Park, Heui-Dong

    2016-09-01

    In this study, Lactobacillus plantarum JH287 was used as a malolactic fermentation starter in Campbell Early wine production. L. plantarum JH287 was first lyophilized, and the malolactic fermentation potential of freeze-dried L. plantarum JH287 was investigated. Different protective media and rehydration conditions were tested to improve the survival rate of freeze-dried L. plantarum JH287. Optimal protective medium contained 10 % sorbitol and 10 % skim milk. The optimal rehydration condition was a 1-h rehydration time conducted in the same protective media, and the combination of these two methods produced a survival rate of 86.37 %. In addition, a 77.71 % survival rate was achieved using freeze-dried samples that were stored at 4 °C for 2 months. Freeze-dried L. plantarum JH287 and Saccharomyces cerevisiae Fermivin were used to inoculate the Campbell Early grape must to decrease its malic acid content. Using this mixed-fermentation method, wine showed a decrease in malic acid content after 9 days of fermentation. GC-MS analysis detected 15 volatile ester compounds in the wine. A sensory evaluation showed that the taste and aroma of mix-fermented wine were better than those of the control that had not been inoculated with L. plantarum JH287.

  6. Effects of egg white protein supplementation on muscle strength and serum free amino acid concentrations.

    Science.gov (United States)

    Hida, Azumi; Hasegawa, Yuko; Mekata, Yuko; Usuda, Mika; Masuda, Yasunobu; Kawano, Hitoshi; Kawano, Yukari

    2012-10-19

    The aim of this study was to evaluate the effects of egg white protein compared to carbohydrate intake prior to exercise on fat free mass (FFM), one repetition maximum (1RM) muscle strength and blood biochemistry in female athletes. Thirty healthy female collegiate athletes were recruited for this study and matched by sport type, body fat percentage and 1RM leg curl muscle strength. Participants were randomly divided into two groups: protein group (15.0 g egg white protein; 75 kcal) and carbohydrate group (17.5 g maltodextrin, 78 kcal). Supplements were administered daily at the same time in a double-blind manner prior to training during an 8-week period. Measurements were performed before and after the 8-week regimen. The mean dietary energy intake did not change throughout the study period. FFM and 1RM assessments (i.e., leg curl, leg extension, squat, and bench press) increased in both groups. Furthermore, serum urea and serum citrulline levels after the 8-week regimen increased significantly only in the protein group. Our findings indicated that compared to the carbohydrate supplement, the protein supplement was associated with some changes in protein metabolites but not with changes in body composition or muscle strength.

  7. Effects of Egg White Protein Supplementation on Muscle Strength and Serum Free Amino Acid Concentrations

    Directory of Open Access Journals (Sweden)

    Yukari Kawano

    2012-10-01

    Full Text Available The aim of this study was to evaluate the effects of egg white protein compared to carbohydrate intake prior to exercise on fat free mass (FFM, one repetition maximum (1RM muscle strength and blood biochemistry in female athletes. Thirty healthy female collegiate athletes were recruited for this study and matched by sport type, body fat percentage and 1RM leg curl muscle strength. Participants were randomly divided into two groups: protein group (15.0 g egg white protein; 75 kcal and carbohydrate group (17.5 g maltodextrin, 78 kcal. Supplements were administered daily at the same time in a double-blind manner prior to training during an 8-week period. Measurements were performed before and after the 8-week regimen. The mean dietary energy intake did not change throughout the study period. FFM and 1RM assessments (i.e., leg curl, leg extension, squat, and bench press increased in both groups. Furthermore, serum urea and serum citrulline levels after the 8-week regimen increased significantly only in the protein group. Our findings indicated that compared to the carbohydrate supplement, the protein supplement was associated with some changes in protein metabolites but not with changes in body composition or muscle strength.

  8. Effects of Egg White Protein Supplementation on Muscle Strength and Serum Free Amino Acid Concentrations

    Science.gov (United States)

    Hida, Azumi; Hasegawa, Yuko; Mekata, Yuko; Usuda, Mika; Masuda, Yasunobu; Kawano, Hitoshi; Kawano, Yukari

    2012-01-01

    The aim of this study was to evaluate the effects of egg white protein compared to carbohydrate intake prior to exercise on fat free mass (FFM), one repetition maximum (1RM) muscle strength and blood biochemistry in female athletes. Thirty healthy female collegiate athletes were recruited for this study and matched by sport type, body fat percentage and 1RM leg curl muscle strength. Participants were randomly divided into two groups: protein group (15.0 g egg white protein; 75 kcal) and carbohydrate group (17.5 g maltodextrin, 78 kcal). Supplements were administered daily at the same time in a double-blind manner prior to training during an 8-week period. Measurements were performed before and after the 8-week regimen. The mean dietary energy intake did not change throughout the study period. FFM and 1RM assessments (i.e., leg curl, leg extension, squat, and bench press) increased in both groups. Furthermore, serum urea and serum citrulline levels after the 8-week regimen increased significantly only in the protein group. Our findings indicated that compared to the carbohydrate supplement, the protein supplement was associated with some changes in protein metabolites but not with changes in body composition or muscle strength. PMID:23201768

  9. In vitro maintenance of spermatogenesis in Xenopus laevis testis explants cultured in serum-free media

    International Nuclear Information System (INIS)

    Risley, M.S.; Miller, A.; Bumcrot, D.A.

    1987-01-01

    Spermatogenesis has been maintained for extended periods in Xenopus laevis testis explants cultured in serum-free media supplemented with bovine serum albumin, insulin, transferrin, follicle-stimulating hormone, dihydrotestosterone, testosterone, retinol, ascorbate, and tocopherol. The organization of the testis fragments was maintained for 28 days, and all stages of development were present throughout the culture period. 3 H-Thymidine-labeled secondary (Type B) spermatogonia developed in 28 days into spermatids at the acrosomal vesicle stage whereas labeled zygotene spermatocytes became mature spermatids in 28 days. Spermatogonial proliferation also continued in vitro for 28 days. Germ cell differentiation was not dependent upon exogenous testosterone, ascorbate, or tocopherol since 3 H-labeled spermatogonia became mature spermatids in testes cultured 35 days in media lacking these supplements. Autoradiography demonstrated that 55% of the luminal sperm present in explants cultured 10 days had differentiated in vitro. Sperm from testes cultured 10-35 days were similar to sperm from freshly dissected testes with regard to motility and fecundity, and eggs fertilized with sperm from explant cultures developed normally into swimming tadpoles. The results demonstrate the feasibility of maintaining vertebrate spermatogenesis in culture and suggest that in vitro analysis of Xenopus spermatogenesis using defined media may provide important insights into the evolution of regulatory mechanisms in spermatogenesis

  10. Glucose and phosphate modulation of intracellular 45Ca incorporated into pancreatic islets during culture in the absence and presence of serum

    International Nuclear Information System (INIS)

    Bergsten, P.

    1985-01-01

    The effects of glucose and phosphate on the intracellular 45 Ca content were measured in β cell-rich pancreatic islets cultured in media containing or lacking serum. Irrespective of the glucose and serum concentrations there were no or very small increments of 45 Ca contents when phosphate was raised from 0.8 to 5.8 mM during culture for 1 day. However, after 7 days of culture in serum-free medium there was a massive accumulation of 45 Ca in the islets in response to the higher phosphate concentration. Glucose markedly reduced, and serum eliminated, the extensive accumulation probably due to increased cell viability. In the cells cultured in the presence of serum, raising the glucose concentration from 1.0 to 5.5 mM resulted in an increased incorporation of 45 Ca. This effect was particularly pronounced after culture for 7 days in 5.8 mM phosphate. A further increase of glucose to 20 mM reduced the 45 Ca content. The results are consistent with the concept that glucose both stimulates 45 Ca uptake into different β-cell pools and degranulates the cell with associated loss of intracellular calcium from the granular calcium pool. (author)

  11. Successful long-term preservation of rat sperm by freeze-drying.

    Directory of Open Access Journals (Sweden)

    Takehito Kaneko

    Full Text Available BACKGROUND: Freeze-drying sperm has been developed as a new preservation method where liquid nitrogen is no longer necessary. An advantage of freeze-drying sperm is that it can be stored at 4 °C and transported at room temperature. Although the successful freeze-drying of sperm has been reported in a number of animals, the possibility of long-term preservation using this method has not yet been studied. METHODOLOGY/PRINCIPAL FINDINGS: Offspring were obtained from oocytes fertilized with rat epididymal sperm freeze-dried using a solution containing 10 mM Tris and 1 mM EDTA adjusted to pH 8.0. Tolerance of testicular sperm to freeze-drying was increased by pre-treatment with diamide. Offspring with normal fertility were obtained from oocytes fertilized with freeze-dried epididymal sperm stored at 4 °C for 5 years. CONCLUSIONS AND SIGNIFICANCE: Sperm with -SS- cross-linking in the thiol-disulfide of their protamine were highly tolerant to freeze-drying, and the fertility of freeze-dried sperm was maintained for 5 years without deterioration. This is the first report to demonstrate the successful freeze-drying of sperm using a new and simple method for long-term preservation.

  12. Norepinephrine stimulates progesterone production in highly estrogenic bovine granulosa cells cultured under serum-free, chemically defined conditions.

    Science.gov (United States)

    Piccinato, Carla A; Montrezor, Luis H; Collares, Cristhianna A V; Vireque, Alessandra A; Rosa e Silva, Alzira A M

    2012-11-22

    Since noradrenergic innervation was described in the ovarian follicle, the actions of the intraovarian catecholaminergic system have been the focus of a variety of studies. We aimed to determine the gonadotropin-independent effects of the catecholamine norepinephrine (NE) in the steroid hormone profile of a serum-free granulosa cell (GC) culture system in the context of follicular development and dominance. Primary bovine GCs were cultivated in a serum-free, chemically defined culture system supplemented with 0.1% polyvinyl alcohol. The culture features were assessed by hormone measurements and ultrastructural characteristics of GCs. GCs produced increasing amounts of estradiol and pregnenolone for 144h and maintained ultrastructural features of healthy steroidogenic cells. Progesterone production was also detected, although it significantly increased only after 96h of culture. There was a highly significant positive correlation between estradiol and pregnenolone production in high E2-producing cultures. The effects of NE were further evaluated in a dose-response study. The highest tested concentration of NE (10 (-7) M) resulted in a significant increase in progesterone production, but not in estradiol or pregnenolone production. The specificity of NE effects on progesterone production was further investigated by incubating GCs with propranolol (10 (-8) M), a non-selective beta-adrenergic antagonist. The present culture system represents a robust model to study the impact of intrafollicular factors, such as catecholamines, in ovarian steroidogenesis and follicular development. The results of noradrenergic effects in the steroidogenesis of GC have implications on physiological follicular fate and on certain pathological ovarian conditions such as cyst formation and anovulation.

  13. Optimization Of Freeze-Dried Starter For Yogurt By Full Factorial Experimental Design

    Directory of Open Access Journals (Sweden)

    Chen He

    2015-12-01

    Full Text Available With the rapidly development of fermented milk product, it is significant for enhancing the performance of starter culture. This paper not only investigated the influence of anti-freeze factors and freeze-drying protective agents on viable count, freeze-drying survival rate and yield of Lactobacillus bulgaricus (LB and Streptococcus thermophilus (ST, but also optimized the bacteria proportion of freeze-dried starter culture for yogurt by full factorial experimental design. The results showed as following: the freeze-drying protective agents or anti-freeze factors could enhanced survival rate of LB and ST; the freeze-dried LB and ST powders containing both of anti-freeze factors and freeze-drying protective agents had higher viable count and freeze-drying survival rate that were 84.7% and 79.7% respectively; In terms of fermentation performance, the best group of freeze-dried starter for yogurt was the compound of LB3 and ST2.

  14. Free leptin index and PAPP-A: a first trimester maternal serum screening test for pre-eclampsia

    DEFF Research Database (Denmark)

    Hedley, Paula L; Placing, Sophie; Wøjdemann, Karen

    2010-01-01

    BACKGROUND: Prophylaxis with low-dose aspirin may reduce the risk of pre-eclampsia (PE) if introduced in first trimester. The performance of first trimester maternal serum screening for PE using free leptin index (fLI) and PAPP-A, where fLI = leptin/leptin soluble receptor was studied. METHODS: F......: First trimester serum samples from 126 PE pregnancies and 289 control pregnancies were studied. fLI and PAPP-A were converted into gestational age and maternal weight independent log MoM values of PAPP-A and fLI. The screening performance of markers was studied by receiver......-operator-characteristics curves. The performance of population screening was estimated by Monte Carlo simulation. RESULTS: fLI was significantly (p controls [mean log MoM -0.0368 (SD: 0.3132)] and PAPP-A was significantly (p ....0133 (SD: 0.2661)] compared to controls [mean log MoM 0.0474 (SD: 0.2521)] in PE pregnancies. There was no correlation between fLI and PAPP-A in control or PE pregnancies. Combined fLI and PAPP-A screening for PE had estimated population detection rates of 22% and 35% for false positives rates of 6% and 12...

  15. Serum testosterone concentration in chloroquine- treated rats ...

    African Journals Online (AJOL)

    ONOS

    2010-07-05

    Jul 5, 2010 ... The effects of ascorbic acid (vitamin C) and alpha-tocopherol (vitamin E) were studied on serum testosterone ... chloroquine are probably mediated via the generation of free radicals. ... Effects of ascorbic acid and alpha-tocopherol on serum testosterone concentration in chloroquine-treated rats. Groups.

  16. Ratio of serum free triiodothyronine to free thyroxine in Graves' hyperthyroidism and thyrotoxicosis caused by painless thyroiditis.

    Science.gov (United States)

    Yoshimura Noh, Jaeduk; Momotani, Naoko; Fukada, Shuji; Ito, Koichi; Miyauchi, Akira; Amino, Nobuyuki

    2005-10-01

    The serum T3 to T4 ratio is a useful indicator for differentiating destruction-induced thyrotoxicosis from Graves' thyrotoxicosis. However, the usefulness of the serum free T3 (FT3) to free T4 (FT4) ratio is controversial. We therefore systematically evaluated the usefulness of this ratio, based on measurements made using two widely available commercial kits in two hospitals. Eighty-two untreated patients with thyrotoxicosis (48 patients with Graves' disease and 34 patients with painless thyroiditis) were examined in Kuma Hospital, and 218 patients (126 with Graves' disease and 92 with painless thyroiditis) and 66 normal controls were examined in Ito Hospital. The FT3 and FT4 values, as well as the FT3/FT4 ratios, were significantly higher in the patients with Graves' disease than in those with painless thyroiditis in both hospitals, but considerable overlap between the two disorders was observed. Receiver operating characteristic (ROC) curves for the FT3 and FT4 values and the FT3/FT4 ratios of patients with Graves' disease and those with painless thyroiditis seen in both hospitals were prepared, and the area under the curves (AUC), the cut-off points for discriminating Graves' disease from painless thyroiditis, the sensitivity, and the specificity were calculated. AUC and sensitivity of the FT(3)/FT(4) ratio were smaller than those of FT(3) and FT(4) in both hospitals. The patients treated at Ito hospital were then divided into 4 groups according to their FT4 levels (A: 2.3 approximately 5.4 ng/dl), and the AUC, cut-off points, sensitivity, and specificity of the FT(3)/FT(4) ratios were calculated. The AUC and sensitivity of each group increased with the FT4 levels (AUC: 57.8%, 72.1%, 91.1%, and 93.4%, respectively; sensitivity: 62.6%, 50.0%, 77.8%, and 97.0%, respectively). The means +/- SE of the FT3/FT4 ratio in the Graves' disease groups were 3.1 +/- 0.22, 3.1 +/- 0.09, 3.2 +/- 0.06, and 3.1 +/- 0.07, respectively, versus 2.9 +/- 0.1, 2.6 +/- 0.07, 2.5 +/- 0

  17. A rapidly negotiable first-stage nuclear freeze

    International Nuclear Information System (INIS)

    Anon.

    1988-01-01

    This paper reports on the objective of a nuclear freeze which is to slow down or stop the so-far inexorable development and deployment of more and more (read destructive and deadly) nuclear warheads. The essential notion is not new. The proposed treaty for a comprehensive ban on nuclear tests that was very nearly negotiated in 1959 was perhaps the first serious effort to obtain a nuclear freeze, albeit a partial one. Growing concern about the nuclear arms race has led to greatly increased interest in much broader and more effective freezes. A comprehensive nuclear freeze, one that would stop all stages in the manufacture, testing, and deployment of nuclear warheads, would clearly be very desirable and have a great impact. It would not, however, deal with the other worrisome aspects of nuclear weapons, which is the very large number of such weapons that already exist

  18. Mechanisms of deterioration of nutrients. [improved quality of freeze-dried foods

    Science.gov (United States)

    Karel, M.; Flink, J. M.

    1978-01-01

    Methods for improving the quality of freeze-dried foods were investigated. Areas discussed include: (1) microstructure of freeze-dried systems, (2) structural changes in freeze-dried systems, (3) artificial food matrices, and (4) osmotic preconcentration to yield improved freeze-dried products.

  19. Cost-Effectiveness of the Freeze-All Policy.

    Science.gov (United States)

    Roque, Matheus; Valle, Marcello; Guimarães, Fernando; Sampaio, Marcos; Geber, Selmo

    2015-08-01

    To evaluate the cost-effectiveness of freeze-all cycles when compared to fresh embryo transfer. This was an observational study with a cost-effectiveness analysis. The analysis consisted of 530 intracytoplasmic sperm injection (ICSI) cycles in a private center in Brazil between January 2012 and December 2013. A total of 530 intracytoplasmic sperm injection (ICSI) cycles - 351 fresh embryo transfers and 179 freeze-all cycles - with a gonadotropin-releasing hormone (GnRH) antagonist protocol and day 3 embryo transfers. The pregnancy rate was 31.1% in the fresh group and 39.7% in the freeze-all group. We performed two scenario analyses for costs. In scenario 1, we included those costs associated with the ICSI cycle (monitoring during controlled ovarian stimulation [COS], oocyte retrieval, embryo transfer, IVF laboratory, and medical costs), embryo cryopreservation of supernumerary embryos, hormone measurements during COS and endometrial priming, medication use (during COS, endometrial priming, and luteal phase support), ultrasound scan for frozen- thawed embryo transfer (FET), obstetric ultrasounds, and miscarriage. The total cost (in USD) per pregnancy was statistically lower in the freeze-all cycles (19,156.73 ± 1,732.99) when compared to the fresh cycles (23,059.72 ± 2,347.02). Even in Scenario 2, when charging all of the patients in the freeze-all group for cryopreservation (regardless of supernumerary embryos) and for FET, the fresh cycles had a statistically significant increase in treatment costs per ongoing pregnancy. The results presented in this study suggest that the freeze-all policy is a cost-effective strategy when compared to fresh embryo transfer.

  20. Serum protein electrophoresis values for free-ranging and zoo-based koalas (Phascolarctos cinereus).

    Science.gov (United States)

    Pye, Geoffrey W; Ellis, William; Fitzgibbon, Sean; Opitz, Brian; Keener, Laura; Arheart, Kristopher L; Cray, Carolyn

    2012-03-01

    In a clinical setting, especially with species of special interest, it is important to use all clinical pathology testing options for general health monitoring and diagnosis. Protein electrophoresis (EPH) has previously been shown to be an important adjunct tool in veterinary medicine. Serum samples from 18 free-ranging and 12 zoo-based koalas (Phascolarctos cinereus) were subject to EPH analysis. Significant differences were found between the two groups for the following values: total protein, albumin, beta globulins, and albumin-globulin ratio (P < 0.05). By using the combined data, the minimum-maximum values for the EPH fractions were as follows: total protein 5.0-7.8 g/dl, albumin 2.8-4.7 g/dl, alpha-1 globulins 0.5-1.1 g/dl, alpha-2 globulins 0.3-0.7 g/dl, beta globulins 0.4-1.0 g/dl, gamma globulins 0.2-1.0 g/dl, and albumin-globulin ratio 1.0-2.1.

  1. Metabolic changes in Avena sativa crowns recovering from freezing.

    Directory of Open Access Journals (Sweden)

    Cynthia A Henson

    Full Text Available Extensive research has been conducted on cold acclimation and freezing tolerance of fall-sown cereal plants due to their economic importance; however, little has been reported on the biochemical changes occurring over time after the freezing conditions are replaced by conditions favorable for recovery and growth such as would occur during spring. In this study, GC-MS was used to detect metabolic changes in the overwintering crown tissue of oat (Avena sativa L. during a fourteen day time-course after freezing. Metabolomic analysis revealed increases in most amino acids, particularly proline, 5-oxoproline and arginine, which increased greatly in crowns that were frozen compared to controls and correlated very significantly with days after freezing. In contrast, sugar and sugar related metabolites were little changed by freezing, except sucrose and fructose which decreased dramatically. In frozen tissue all TCA cycle metabolites, especially citrate and malate, decreased in relation to unfrozen tissue. Alterations in some amino acid pools after freezing were similar to those observed in cold acclimation whereas most changes in sugar pools after freezing were not. These similarities and differences suggest that there are common as well as unique genetic mechanisms between these two environmental conditions that are crucial to the winter survival of plants.

  2. Applications of Simulator Freeze to Carrier Glideslope Tracking Instruction.

    Science.gov (United States)

    1982-07-01

    Showing Datum Bars and Meatball . .. .. .. ... .. ... .... 19 4 Freezes Per Trial Averaged Across Freeze Conditions and Across 4-Trial Blocks of Training...algorithm linearly increased the criterion in meatball units from 1.0 at 6000 feet from the ramp to 1.5 at the ramp. "Freezes" did not occur beyond 6000

  3. Effects of sugar alcohol and proteins on the survival of Lactobacillus bulgaricus LB6 during freeze drying.

    Science.gov (United States)

    Chen, He; Chen, Shiwei; Chen, Hongli; Wu, Yanyan; Shu, Guowei

    2015-01-01

    Lactobacillus bulgaricus LB6 is a bacterium which was selected in the commercial yoghurt with high angiotensin converting enzyme (ACE) inhibitory activity. Preparation of concentrated starter cultures via freeze drying is of practical importance to dairy and food industries. We optimized the optimal sugar alcohol and proteins for Lactobacillus bulgaricus LB6 during the process of freeze drying using a Plackett-Burman design. In our initial tests survival rate and the number of viable cells were associated with the type of lyoprotectant used and so our optimization protocol focused on increasing survival rate. Substances that had previously had a protective effect during freeze drying were investigated, for example: mannitol, sorbitol, xylitol, meso-erythritol, lactitol, whey protein isolate 90, bovine serum albumin, and whey protein concentrate 80 and soy protein isolate 70. We found that the optimum sugar alcohol and proteins for survival of Lactobacillus bulgaricus LB6 were whey protein concentrate (p = 0.0040 for survival rate), xylitol (p = 0.0067 for survival rate) and sorbitol (p = 0.0073 for survival rate), they showed positive effect (whey protein concentrate and sorbitol) or negative effect (xylitol). The effectiveness of three chosen sugar alcohols and protein implied that they could be used as lyoprotectant for Lactobacillus bulgaricus LB6 in the further research, the optimal composition of sugar alcohol and protein for the lyoprotectant use must be established.

  4. Mesenchymal stem cell-conditioned medium accelerates skin wound healing: An in vitro study of fibroblast and keratinocyte scratch assays

    International Nuclear Information System (INIS)

    Walter, M.N.M.; Wright, K.T.; Fuller, H.R.; MacNeil, S.; Johnson, W.E.B.

    2010-01-01

    We have used in vitro scratch assays to examine the relative contribution of dermal fibroblasts and keratinocytes in the wound repair process and to test the influence of mesenchymal stem cell (MSC) secreted factors on both skin cell types. Scratch assays were established using single cell and co-cultures of L929 fibroblasts and HaCaT keratinocytes, with wound closure monitored via time-lapse microscopy. Both in serum supplemented and serum free conditions, wound closure was faster in L929 fibroblast than HaCaT keratinocyte scratch assays, and in co-culture the L929 fibroblasts lead the way in closing the scratches. MSC-CM generated under serum free conditions significantly enhanced the wound closure rate of both skin cell types separately and in co-culture, whereas conditioned medium from L929 or HaCaT cultures had no significant effect. This enhancement of wound closure in the presence of MSC-CM was due to accelerated cell migration rather than increased cell proliferation. A number of wound healing mediators were identified in MSC-CM, including TGF-β1, the chemokines IL-6, IL-8, MCP-1 and RANTES, and collagen type I, fibronectin, SPARC and IGFBP-7. This study suggests that the trophic activity of MSC may play a role in skin wound closure by affecting both dermal fibroblast and keratinocyte migration, along with a contribution to the formation of extracellular matrix.

  5. Free convective flow of a stratified fluid through a porous medium bounded by a vertical plane

    Directory of Open Access Journals (Sweden)

    H. K. Mondal

    1994-01-01

    Full Text Available Steady two-dimensional free convection flow of a thermally stratified viscous fluid through a highly porous medium bounded by a vertical plane surface of varying temperature, is considered. Analytical expressions for the velocity, temperature and the rate of heat transfer are obtained by perturbation method. Velocity distribution and rate of heat transfer for different values of parameters are shown in graphs. Velocity distribution is also obtained for certain values of the parameters by integrating the coupled differential equations by Runge-Kutta method and compared with the analytical solution. The chief concern of the paper is to study the effect of equilibrium temperature gradient on the velocity and the rate of heat transfer.

  6. Serum complexed and free prostate specific antigen levels are lower in female elite athletes in comparison to control women [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Emma Eklund

    2017-07-01

    Full Text Available Background: We hypothesize that prostate specific antigen (PSA, a protein that it is under regulation by androgens, may be differentially expressed in female elite athletes in comparison to control women. Methods: We conducted a cross-sectional study of 106 female athletes and 114 sedentary age-matched controls.  Serum from these women was analyzed for complexed prostate specific antigen (cPSA and free prostate specific antigen (fPSA, by fifth generation assays with limits of detection of around 6 and 140 fg/mL, respectively.  A panel of estrogens, androgens and progesterone in the same serum was also quantified by tandem mass spectrometry.  Results: Both components of serum PSA (cPSA and fPSA were lower in the elite athletes vs the control group (P=0.033 and 0.013, respectively.  Furthermore, estrone (p=0.003 and estradiol (p=0.004 were significantly lower, and dehydroepiandrosterone  (p=0.095 and 5-androstene-3β, 17β-diol (p=0.084 tended to be higher in the athletes vs controls. Oral contraceptive use was similar between groups and significantly associated with increased cPSA and fPSA in athletes (p= 0.046 and 0.009, respectively.  PSA fractions were not significantly associated with progesterone changes. The Spearman correlation between cPSA and fPSA in both athletes and controls was 0.75 (P < 0.0001 and 0.64 (P < 0.0001, respectively.  Conclusions: Elite athletes have lower complexed and free PSA, higher levels of androgen precursors and lower levels of estrogen in their serum than sedentary control women. Abbreviations: cPSA, complexed PSA; fPSA, free PSA; PCOS, polycystic ovarian syndrome; E1, estrone; E2, estradiol; DHEA, dehydroepiandrosterone, Testo, testosterone; DHT, dihydrotestosterone; PROG, progesterone; Delta 4, androstenedione; Delta 5, androst-5-ene-3β, 17β-diol; BMD, body mineral density; LLOQ, lower limit of quantification; ULOQ, upper limit of quantification; LOD, limit of detection; ACT, α1-antichymotrypsin

  7. The stability of complement-mediated bactericidal activity in human serum against Salmonella.

    Directory of Open Access Journals (Sweden)

    Colette M O'Shaughnessy

    Full Text Available The complement cascade includes heat-labile proteins and care is required when handling serum in order to preserve its functional integrity. We have previously used a whole human serum bactericidal assay to show that antibody and an intact complement system are required in blood for killing of invasive isolates of Salmonella. The aim of the present study was to evaluate the conditions under which human serum can be stored and manipulated while maintaining complement integrity. Serum bactericidal activity against Salmonella was maintained for a minimum of 35 days when stored at 4°C, eight days at 22°C and 54 hours at 37°C. Up to three freeze-thaw cycles had no effect on the persistence of bactericidal activity and hemolytic complement assays confirmed no effect on complement function. Delay in the separation of serum for up to four days from clotted blood stored at 22°C did not affect bactericidal activity. Dilution of serum resulted in an increased rate of loss of bactericidal activity and so serum should be stored undiluted. These findings indicate that the current guidelines concerning manipulation and storage of human serum to preserve complement integrity and function leave a large margin for safety with regards to bactericidal activity against Salmonella. The study provides a scheme for determining the requirements for serum handling in relation to functional activity of complement in other systems.

  8. Freeze block testing of buried waste lines

    International Nuclear Information System (INIS)

    Robbins, E.D.; Willi, J.C.

    1976-01-01

    An investigation was conducted to demonstrate application of freeze blocking in waste transfer lines such that a hydrostatic pressure test can be applied. A shop test was conducted on a 20-foot length, 3-inch schedule 40, carbon steel pipe using a coolant of dry ice and Freon. The positive results from these tests prompted a similar employment of the freeze block method in hydrostatic pressure testing the feed inlet leading to 241-S-101 Waste Tank. This pipeline is a 3-inch schedule 10, stainless steel pipe approximately 800 feet long. The freeze block was formed near the lower end of the pipe as it entered the 101-S Waste Tank and a pressure hold test was applied to this pipeline. This test proved the integrity of the pipeline in question, and demonstrated the validity of freeze blocking an open-ended pipeline which could not be hydrotested in other conventional ways. The field demonstration facility, costing $30,200 was completed late in 1975

  9. IgD multiple myeloma: Clinical, biological features and prognostic value of the serum free light chain assay.

    Science.gov (United States)

    Djidjik, R; Lounici, Y; Chergeulaïne, K; Berkouk, Y; Mouhoub, S; Chaib, S; Belhani, M; Ghaffor, M

    2015-09-01

    IgD multiple myeloma (MM) is a rare subtype of myeloma, it affects less than 2% of patients with MM. To evaluate the clinical and prognostic attributes of serum free light chains (sFLCs) analysis, we examined 17 cases of IgD MM. From 1998 to 2012, we obtained 1250 monoclonal gammapathies including 590 multiple myeloma and 17 patients had IgD MM. With preponderance of men patients with a mean age at diagnosis of: 59±12years. Patients with IgD MM have a short survival (Median survival=9months). The presenting features included: bone pain (75%), lymphadenopathy (16%), hepatomegaly (25%), splenomegaly (8%), associated AL amyloidosis (6%), renal impairment function (82%), infections (47%), hypercalcemia (37%) and anemia (93%). Serum electrophoresis showed a subtle M-spike (Mean=13.22±10g/L) in all patients associated to a hypogammaglobulinemia. There was an over-representation of Lambda light chain (65%); high serum β2-microglobulin in 91% and Bence Jones proteinuria was identified in 71%. The median rate of sFLCs κ was 19.05mg/L and 296.75mg/L for sFLCs λ. sFLCR was abnormal in 93% of patients and it showed concordance between baseline sFLCR and the survival (P=0.034). The contribution of FLC assay is crucial for the prognosis of patients with IgD MM. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  10. Pathogen-free, plasma-poor platelet lysate and expansion of human mesenchymal stem cells.

    Science.gov (United States)

    Iudicone, Paola; Fioravanti, Daniela; Bonanno, Giuseppina; Miceli, Michelina; Lavorino, Claudio; Totta, Pierangela; Frati, Luigi; Nuti, Marianna; Pierelli, Luca

    2014-01-27

    Supplements to support clinical-grade cultures of mesenchymal stem cells (MSC) are required to promote growth and expansion of these cells. Platelet lysate (PL) is a human blood component which may replace animal serum in MSC cultures being rich in various growth factors. Here, we describe a plasma poor pathogen-free platelet lysate obtained by pooling 12 platelet (PLT) units, to produce a standardized and safe supplement for clinical-grade expansion of MSC. PL lots were obtained by combining 2 6-unit PLT pools in additive solution (AS) following a transfusional-based procedure including pathogen inactivation (PI) by Intercept technology and 3 cycles of freezing/thawing, followed by membrane removal. Three PI-PL and 3 control PL lots were produced to compare their ability to sustain bone marrow derived MSC selection and expansion. Moreover, two further PL, subjected to PI or not, were also produced starting from the same initial PLT pools to evaluate the impact of PI on growth factor concentration and capacity to sustain cell growth. Additional PI-PL lots were used for comparison with fetal bovine serum (FBS) on MSC expansion. Immunoregulatory properties of PI-PL-generated MSC were documented in vitro by mixed lymphocyte culture (MLC) and peripheral blood mononuclear cells (PBMC) mitogen induced proliferation. PI-PL and PL control lots had similar concentrations of 4 well-described growth factors endowed with MSC stimulating ability. Initial growth and MSC expansion by PI-PL and PL controls were comparable either using different MSC populations or in head to head experiments. Moreover, PI-PL and PL control sustained similar MSC growth of frozen/thawed MSC. Multilineage differentiation of PI-derived and PI-PL-derived MSC were maintained in any MSC cultures as well as their immunoregulatory properties. Finally, no direct impact of PI on growth factor concentration and MSC growth support was observed, whereas the capacity of FBS to sustain MSC expansion in basic

  11. Predictive modeling of freezing and thawing of frost-susceptible soils.

    Science.gov (United States)

    2015-09-01

    Frost depth is an essential factor in design of various transportation infrastructures. In frost : susceptible soils, as soils freezes, water migrates through the soil voids below the freezing line : towards the freezing front and causes excessive he...

  12. Multiple Glass Transitions and Freezing Events of Aqueous Citric Acid

    Science.gov (United States)

    2014-01-01

    Calorimetric and optical cryo-microscope measurements of 10–64 wt % citric acid (CA) solutions subjected to moderate (3 K/min) and slow (0.5 and 0.1 K/min) cooling/warming rates and also to quenching/moderate warming between 320 and 133 K are presented. Depending on solution concentration and cooling rate, the obtained thermograms show one freezing event and from one to three liquid–glass transitions upon cooling and from one to six liquid–glass and reverse glass–liquid transitions, one or two freezing events, and one melting event upon warming of frozen/glassy CA/H2O. The multiple freezing events and glass transitions pertain to the mother CA/H2O solution itself and two freeze-concentrated solution regions, FCS1 and FCS2, of different concentrations. The FCS1 and FCS2 (or FCS22) are formed during the freezing of CA/H2O upon cooling and/or during the freezing upon warming of partly glassy or entirely glassy mother CA/H2O. The formation of two FCS1 and FCS22 regions during the freezing upon warming to our best knowledge has never been reported before. Using an optical cryo-microscope, we are able to observe the formation of a continuous ice framework (IF) and its morphology and reciprocal distribution of IF/(FCS1 + FCS2). Our results provide a new look at the freezing and glass transition behavior of aqueous solutions and can be used for the optimization of lyophilization and freezing of foods and biopharmaceutical formulations, among many other applications where freezing plays a crucial role. PMID:25482069

  13. Application of freeze-drying technology in manufacturing orally disintegrating films.

    Science.gov (United States)

    Liew, Kai Bin; Odeniyi, Michael Ayodele; Peh, Kok-Khiang

    2016-01-01

    Freeze drying technology has not been maximized and reported in manufacturing orally disintegrating films. The aim of this study was to explore the freeze drying technology in the formulation of sildenafil orally disintegrating films and compare the physical properties with heat-dried orally disintegrating film. Central composite design was used to investigate the effects of three factors, namely concentration of carbopol, wheat starch and polyethylene glycol 400 on the tensile strength and disintegration time of the film. Heat-dried films had higher tensile strength than films prepared using freeze-dried method. For folding endurance, freeze-dried films showed improved endurance than heat-dried films. Moreover, films prepared using freeze-dried methods were thicker and had faster disintegration time. Formulations with higher amount of carbopol and starch showed higher tensile strength and thickness whereas formulations with higher PEG 400 content showed better flexibility. Scanning electron microscopy showed that the freeze-dried films had more porous structure compared to the heat-dried film as a result of the release of water molecule from the frozen structure when it was subjected to freeze drying process. The sildenafil film was palatable. The dissolution profiles of freeze-dried and heat-dried films were similar to Viagra® with f2 of 51.04 and 65.98, respectively.

  14. Three-Dimensional Microstructure of Biological Tissues during Freezing and Thawing

    Science.gov (United States)

    Ishiguro, Hiroshi; Horimizu, Takashi; Kataori, Akinobu; Kajigaya, Hiroshi

    Three-dimensional behavior of ice crystals and cells during the freezing and thawing of biological tissues was investigated microscopically in real time by using a confocal laser scanning microscope(CLSM) and a fluorescent dye, acridine orange (AO). Fresh tender meat (2nd pectoral muscles) of chicken was stained with the AO in physiological saline to distinguish ice crystals and cells by their different colors, and then frozen and thawed under two different thermal protocols: a) slow-cooling and rapid-warming and b) rapid-cooling and rapid-warming. The CLSM noninvasively produced optical tomograms of the tissues to clarify the pattern of freezing, morphology of ice crystals in the tissues, and the interaction between ice crystals and cells. Also, the tissues were morphologically investigated by pathological means after the freezing and thawing. Typical freezing pattern during the slow-cooling was extracellular-freezing, and those during the rapid-cooling were extracellular-freezing and intracellular freezing with a lot of fine ice crystals in the cells. Cracks caused by the extracellular and intracellular ice crystals remained in the muscle tissues after the thawing. The results obtained by using the CLSM/dye method were consistent with pathologically morphological changes in the tissues through freezing and thawing.

  15. Low cycle fatigue behavior in a medium-carbon carbide-free bainitic steel

    Energy Technology Data Exchange (ETDEWEB)

    Kang, J. [State Key Laboratory of Metastable Materials Science and Technology, Yanshan University, Qinhuangdao 066004 (China); Zhang, F.C., E-mail: zfc@ysu.edu.cn [State Key Laboratory of Metastable Materials Science and Technology, Yanshan University, Qinhuangdao 066004 (China); Long, X.Y. [State Key Laboratory of Metastable Materials Science and Technology, Yanshan University, Qinhuangdao 066004 (China); Lv, B. [College of Environmental and Chemical Engineering, Yanshan University, Qinhuangdao 066004 (China)

    2016-06-01

    In the paper, different morphologies of bainite were obtained through isothermal quenching at 320 °C and 395 °C in a medium-carbon carbide-free bainitic steel. The cyclic deformation mechanism was explored by using low cycle fatigue testing. The volume fraction of retained austenite was measured by X-ray diffraction and the space partitioning of the solute atoms was constructed by three-dimensional atom probe. Results showed that the fatigue life at 320 °C was always higher than that at 395 °C under low and high total strain amplitude. The cyclic softening at the early fatigue stage increased the plastic strain of the sample which was responsible for the reduction of the fatigue life at 395 °C. Strain-induced retained austenite to martensite contributed to initial cyclic hardening, but almost having no effect on the subsequent cyclic stable/softening behaviors. The finer bainitic ferrite sheaves obtained at 320 °C changed the small fatigue crack propagation direction and delayed the crack propagation rate, which was beneficial for the fatigue properties. In addition, the substitutional atoms did not redistribute between the retained austenite and bainitic ferrite before and after cyclic deformation.

  16. Immobilization of sericin molecules via amorphous carbon plasma modified-polystyrene dish for serum-free culture

    Energy Technology Data Exchange (ETDEWEB)

    Tunma, Somruthai [The Graduate School, Chiang Mai University, 239 Huay Kaew Road, Muang District, Chiang Mai 50200 (Thailand); Thailand Center of Excellence in Physics (ThEP), 239 Huay Kaew Road, Muang District, Chiang Mai 50200 (Thailand); Song, Doo-Hoon [Research Center for Orofacial Hard Tissue Regeneration, College of Dentistry, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Kim, Si-Eun; Kim, Kyoung-Nam [Research Center for Orofacial Hard Tissue Regeneration, College of Dentistry, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Department and Research Institute of Dental Biomaterials and Bioengineering, College of Dentistry, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Han, Jeon-Geon [Center for Advanced Plasma Surface Technology, Sungkyunkwan University, 300 Chunchun-dong, Jangan-gu, Suwon 440-746 (Korea, Republic of); Boonyawan, Dheerawan [Thailand Center of Excellence in Physics (ThEP), 239 Huay Kaew Road, Muang District, Chiang Mai 50200 (Thailand); Department of Physics and Materials Science, Faculty of Science, Chiang Mai University, 239 Huay Kaew Road, Muang District, Chiang Mai 50200 (Thailand)

    2013-10-15

    In this study, we focused on sericin hydrolysates, originating from silkworm used in serum-free human bone marrow-derived mesenchymal stem cells (hBM-MSCs) culture. We reported the effect of a covalent linkage between a bioactive protein molecule and polystyrene dish surface via a carbon intermediate layer which can slow down the release rate of protein compounds into the phosphate buffer saline (PBS) solution. Films of amorphous carbon (a-C) and functionalized-carbon were deposited on PS culture dish surfaces by using a DC magnetron sputtering system and RF PECVD system. We found that a-C based-films can increase the hydrophilicity and biocompatibility of polystyrene (PS) dishes, especially a-C films and a-C:N{sub 2} films showed good attachment of hBM-MSCs at 24 h. However, in the case of silica surface (a-C:SiO{sub x} films), the cells showed a ragged and unattached boundary resulting from the presence of surface silanol groups. For the UV–vis absorbance, all carbon modified-PS dishes showed a lower release rate of sericin molecules into PBS solution than PS control. This revealed that the functionalized carbon could be enhanced by specific binding properties with given molecules. The carbon-coated PS dishes grafting with sericin protein were used in a serum-free condition. We also found that hBM-MSCs have higher percentage of proliferated cells at day 7 for the modified dishes with carbon films and coated with sericin than the PS control coated with sericin. The physical film properties were measured by atomic force microscopy (AFM), scanning electron microscope (SEM) and contact angle measurement. The presence of -NH{sub 2} groups of sericin compounds on the PS dish was revealed by Fourier transform infrared spectroscopy (FTIR). The stability of covalent bonds of sericin molecules after washing out ungrafted sericin was confirmed by X-ray photoelectron spectroscopy (XPS).

  17. Protein brownian rotation at the glass transition temperature of a freeze-concentrated buffer probed by superparamagnetic nanoparticles.

    Science.gov (United States)

    Eloi, J-C; Okuda, M; Jones, S E Ward; Schwarzacher, W

    2013-06-18

    For applications from food science to the freeze-thawing of proteins it is important to understand the often complex freezing behavior of solutions of biomolecules. Here we use a magnetic method to monitor the Brownian rotation of a quasi-spherical cage-shaped protein, apoferritin, approaching the glass transition Tg in a freeze-concentrated buffer (Tris-HCl). The protein incorporates a synthetic magnetic nanoparticle (Co-doped Fe3O4 (magnetite)). We use the magnetic signal from the nanoparticles to monitor the protein orientation. As T decreases toward Tg of the buffer solution the protein's rotational relaxation time increases exponentially, taking values in the range from a few seconds up to thousands of seconds, i.e., orders of magnitude greater than usually accessed, e.g., by NMR. The longest relaxation times measured correspond to estimated viscosities >2 MPa s. As well as being a means to study low-temperature, high-viscosity environments, our method provides evidence that, for the cooling protocol used, the following applies: 1), the concentration of the freeze-concentrated buffer at Tg is independent of its initial concentration; 2), little protein adsorption takes place at the interface between ice and buffer; and 3), the protein is free to rotate even at temperatures as low as 207 K. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  18. Norepinephrine stimulates progesterone production in highly estrogenic bovine granulosa cells cultured under serum-free, chemically defined conditions

    Directory of Open Access Journals (Sweden)

    Piccinato Carla A

    2012-11-01

    Full Text Available Abstract Background Since noradrenergic innervation was described in the ovarian follicle, the actions of the intraovarian catecholaminergic system have been the focus of a variety of studies. We aimed to determine the gonadotropin-independent effects of the catecholamine norepinephrine (NE in the steroid hormone profile of a serum-free granulosa cell (GC culture system in the context of follicular development and dominance. Methods Primary bovine GCs were cultivated in a serum-free, chemically defined culture system supplemented with 0.1% polyvinyl alcohol. The culture features were assessed by hormone measurements and ultrastructural characteristics of GCs. Results GCs produced increasing amounts of estradiol and pregnenolone for 144h and maintained ultrastructural features of healthy steroidogenic cells. Progesterone production was also detected, although it significantly increased only after 96h of culture. There was a highly significant positive correlation between estradiol and pregnenolone production in high E2-producing cultures. The effects of NE were further evaluated in a dose–response study. The highest tested concentration of NE (10 (−7 M resulted in a significant increase in progesterone production, but not in estradiol or pregnenolone production. The specificity of NE effects on progesterone productio n was further investigated by incubating GCs with propranolol (10 (−8 M, a non-selective beta-adrenergic antagonist. Conclusions The present culture system represents a robust model to study the impact of intrafollicular factors, such as catecholamines, in ovarian steroidogenesis and follicular development. The results of noradrenergic effects in the steroidogenesis of GC have implications on physiological follicular fate and on certain pathological ovarian conditions such as cyst formation and anovulation.

  19. Effect of Antioxidants and Apoptosis Inhibitors on Cryopreservation of Murine Germ Cells Enriched for Spermatogonial Stem Cells.

    Directory of Open Access Journals (Sweden)

    Seung-Jung Ha

    Full Text Available Spermatogonial stem cells (SSCs are germline stem cells that serve as the foundation of spermatogenesis to maintain fertility throughout a male's lifetime. To treat male infertility using stem cell banking systems and transplantation, it is important to be able to preserve SSCs for long periods of time. Therefore, this study was conducted to develop an optimal cryopreservation protocol for SSCs using antioxidants and apoptosis inhibitors in freezing medium. No differences were observed compared to controls when SSCs were cryopreserved in the presence of apoptosis inhibitors by themselves. However, mouse germ cells cryopreserved in basal medium containing the antioxidant hypotaurine (14 mM resulted in significantly greater proliferation potential and mitochondrial activity. Furthermore, treatment groups with combinations containing 200 mM trehalose and 14 mM hypotaurine showed higher proliferation rates compared to controls. In addition, several serum free conditions were evaluated for SSC cryopreservation. Treatment media containing 10% or 20% knockout serum replacement resulted in similar cryopreservation results compared to media containing FBS. SSC transplantation was also performed to confirm the functionality of SSCs frozen in 14 mM hypotaurine. Donor SSCs formed normal spermatogenic colonies and sperm in the recipient testis. These data indicate that inclusion of 14 mM hypotaurine in cryopreservation media is an effective way to efficiently cryopreserve germ cells enriched for SSCs and that knockout serum replacement can replace FBS in germ cell cryopreservation media.

  20. SCREENING OF MEDIUM COMPOUNDS USING A TWO-LEVEL FACTORIAL DESIGN FOR SACCHAROMYCES BOULARDII

    Directory of Open Access Journals (Sweden)

    GUOWEI SHU

    2016-04-01

    Full Text Available Even if the probiotic effect of Saccharomyces boulardii is has been reported, this yeast is rarely used in medium composition. Based on single factor experiment, two-level factorial design was employed to evaluate the effect of carbon sources (sucrose, glucose, nitrogen sources (soy peptone, beef extract, yeast extract, calf serum, malt extract and salts (K2HPO4, KH2PO4, MgSO4, Na2HPO4, NaH2PO4, CaCl2, sodium citrate, sodium glutamate on the growth of S. boulardii. At the same time, the optical density (OD in the medium was measured at 560 nm after 36 h of incubation. The result of two-level factorial design experiment showed that calf serum (p = 0.0214 and sodium citrate (p = 0.0045 are the significant growth factors of S. boulardii, sucrose (p = 0.0861 and malt extract (p = 0.0763 are important factors. In addition, sucrose and sodium citrate showed positive effect on the growth of S. boulardii. However, calf serum and malt extract showed negative effect on the growth. And we determined that the optimum medium composition for S. boulardii was as follow: 37.5 g·L-1 sucrose, 6 g·L-1 calf serum, 6 g·L-1 malt extract, 5 g·L-1 sodium citrate.

  1. Static delectric behavior of charged fluids near freezing

    International Nuclear Information System (INIS)

    Fasolino, A.; Parrinello, M.; Tosi, M.P.

    1978-01-01

    The wavenumber-dependent, static dielectric function of classical charged fluids near freezing is obtained from structural data based on computer simulation or neutron diffraction, and its behavior is connected with the freezing process. (Auth.)

  2. Cryogenic freezing of fresh date fruits for quality preservation during frozen storage

    Directory of Open Access Journals (Sweden)

    Abdullah Alhamdan

    2018-01-01

    Full Text Available Fresh date fruits, especially Barhi cultivar, are favored and widely consumed at the Khalal maturity stage (first color edible stage. These fruits are seasonal and perishable and there is a need for extending their shelf life. This study evaluates two different freezing methods, namely cryogenic freezing using liquid nitrogen and conventional deep freezing on preserving the quality and stability of date fruits (cv. Barhi at Khalal maturity stage. Fresh date fruits (cv. Barhi at Khalal stage were frozen utilizing the two methods. The produced frozen dates were stored under frozen storage conditions for nine months (at −20 °C and −40 °C for the conventional and cryogenic freezing, respectively. Color values, textural properties (hardness, elasticity, chewiness and resilience, and nutrition attributes (enzymes and sugars for fresh dates before freezing and for the frozen dates were measured every three months during the frozen storage. Color values of the frozen dates were affected by the freezing method and the frozen storage period. There are substantial differences in the quality of the frozen fruits in favor of cryogenic freezing compared to the conventional slow freezing. The results revealed a large disparity between the times of freezing of the two methods. The freezing time accounted to 10 min in the cryogenic freezing method, whereas it was 1800 min for the conventional slow freezing system.

  3. Medium effects in direct reactions

    International Nuclear Information System (INIS)

    Karakoc, M; Bertulani, C

    2013-01-01

    We discuss medium corrections of the nucleon-nucleon (NN) cross sections and their influence on direct reactions at intermediate energies ≳50 MeV/nucleon. The results obtained with free NN cross sections are compared with those obtained with a geometrical treatment of Pauli-blocking and Dirac-Bruecker methods. We show that medium corrections may lead to sizable modifications for collisions at intermediate energies and that they are more pronounced in reactions involving weakly bound nuclei.

  4. Experimental analysis and modeling of ultrasound assisted freezing of potato spheres.

    Science.gov (United States)

    Kiani, Hossein; Zhang, Zhihang; Sun, Da-Wen

    2015-09-01

    In recent years, innovative methods such as ultrasound assisted freezing have been developed in order to improve the freezing process. During freezing of foods, accurate prediction of the temperature distribution, phase ratios, and process time is very important. In the present study, ultrasound assisted immersion freezing process (in 1:1 ethylene glycol-water solution at 253.15K) of potato spheres (0.02 m diameter) was evaluated using experimental, numerical and analytical approaches. Ultrasound (25 kHz, 890 W m(-2)) was irradiated for different duty cycles (DCs=0-100%). A finite volume based enthalpy method was used in the numerical model, based on which temperature and liquid fraction profiles were simulated by a program developed using OpenFOAM® CFD software. An analytical technique was also employed to calculate freezing times. The results showed that ultrasound irradiation could decrease the characteristic freezing time of potatoes. Since ultrasound irradiation increased the heat transfer coefficient but simultaneously generated heat at the surface of the samples, an optimum DC was needed for the shortest freezing time which occurred in the range of 30-70% DC. DCs higher than 70% increased the freezing time. DCs lower than 30% did not provide significant effects on the freezing time compared to the control sample. The numerical model predicted the characteristic freezing time in accordance with the experimental results. In addition, analytical calculation of characteristic freezing time exhibited qualitative agreement with the experimental results. As the numerical simulations provided profiles of temperature and water fraction within potatoes frozen with or without ultrasound, the models can be used to study and control different operation situations, and to improve the understanding of the freezing process. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Freeze-drying and related preparation techniques for biological microprobe analysis

    International Nuclear Information System (INIS)

    Wroblewski, R.; Wroblewski, J.; Anniko, M.; Edstroem, L.P.

    1985-01-01

    An X-ray microanalytical and morphological investigation has been carried out on rapidly frozen, freeze-dried or freeze-substituted tissues. A comparison was made between different embedding and polymerization procedures following freeze-substitution and freeze-drying. The investigation also included an analysis of specimens infiltrated, embedded and polymerized by ultraviolet irradiation at low temperatures with Lowicryl HM20. The morphological preservation of Lowicryl embedded tissue was adequate for the identification of different cell structures like nuclei, mitochondria, lysosomes and different types of endoplasmic reticulum. X-ray microanalytical investigation of low temperature embedded material displayed an elemental composition of cells and organelles similar to that found in freeze-dried cyosections. Compared with freeze-dried cryosections, low temperature embedded material could be sectioned for light microscopy and area of interest chosen for further thin sectioning. This is of great importance in work with tissues with complicated morphology and heterogenous cell populations

  6. Oral application of freeze-dried yeast particles expressing the PCV2b Cap protein on their surface induce protection to subsequent PCV2b challenge in vivo.

    Science.gov (United States)

    Patterson, Robert; Eley, Thomas; Browne, Christopher; Martineau, Henny M; Werling, Dirk

    2015-11-17

    Porcine circovirus type 2 (PCV2) is now endemic in every major pig producing country, causing PCV-associated disease (PCVAD), linked with large scale economic losses. Current vaccination strategies are based on the capsid protein of the virus and are reasonably successful in preventing PCVAD but fail to induce sterile immunity. Additionally, vaccinating whole herds is expensive and time consuming. In the present study a "proof of concept" vaccine trial was employed to test the effectiveness of powdered freeze-dried recombinant Saccharomyces cerevisiae yeast stably expressing the capsid protein of PCV2b on its surface as an orally applied vaccine. PCV2-free pigs were given 3 doses of vaccine or left un-vaccinated before challenge with a defined PCV2b strain. Rectal temperatures were measured and serum and faeces samples were collected weekly. At the end of the study, pigs were euthanized, tissue samples taken and tested for PCV2b load by qPCR and immunohistochemistry. The peak of viraemia in sera and faeces of unvaccinated pigs was higher than that of vaccinated pigs. Additionally more sIgA was found in faeces of vaccinated pigs than unvaccinated. Vaccination was associated with lower serum concentrations of TNFα and IL-1β but higher concentrations of IFNα and IFNγ in comparison to the unvaccinated animals. At the end of the trial, a higher viral load was found in several lymphatic tissues and the ileum of unvaccinated pigs in comparison to vaccinated pigs. The difference between groups was especially apparent in the ileum. The results presented here demonstrate a possible use for recombinant S. cerevisiae expressing viral proteins as an oral vaccine against PCV2. A powdered freeze-dried recombinant S. cerevisiae used as an oral vaccine could be mixed with feed and may offer a cheap and less labour intensive alternative to inoculation with the additional advantage that no cooling chain would be required for vaccine transport and storage. Copyright © 2015 The

  7. Heterogeneous freezing of super cooled water droplets in micrometre range- freezing on a chip

    Science.gov (United States)

    Häusler, Thomas; Witek, Lorenz; Felgitsch, Laura; Hitzenberger, Regina; Grothe, Hinrich

    2017-04-01

    A new setup to analyse the freezing behaviour of ice nucleation particles (INPs) dispersed in aqueous droplets has been developed with the aim to analyse ensembles of droplets with sizes in the micrometre range, in which INPs are immersed. Major disadvantages of conventional drop-freezing experiments like varying drop sizes or interactions between the water- oil mixture and the INP, were solved by introducing a unique freezing- chip consisting of an etched and sputtered 15x15x1 mm gold-plated silicon or pure gold film (Pummer et al., 2012; Zolles et al., 2015). Using this chip, isolated micrometre-sized droplets can be generated with sizes similar to droplets in real world clouds. The experimental set-up for drop-freezing experiments was revised and improved by establishing automated process control and image evaluation. We were able to show the efficiency and accuracy of our setup by comparing measured freezing temperatures of different INPs (Snomax®, K- feldspar, birch pollen (Betula pendula) washing water, juniper pollen suspension (Juniperus communis) and ultrapure water) with already published results (Atkinson et al., 2013; Augustin et al., 2013; Pruppacher and Klett, 1997; Pummer et al., 2012; Wex et al., 2015; Zolles et al., 2015). Comparison of our measurements with literature data show the important impact of droplet size, INP concentration and number of active sites on the T50 values. Here, the new set-up exhibits its strength in reproducibility and accuracy which is due to the defined and isolated droplets. Finally, it opens a temperature window down to -37˚ C for freezing experiments which was not accessible with former traditional approaches .Atkinson, J. D., Murray, B. J., Woodhouse, M. T., Whale, T. F., Baustian, K. J., Carslaw, K. S., Dobbie, S., O'Sullivan, D., and Malkin, T. L.: The importance of feldspar for ice nucleation by mineral dust in mixed-phase clouds (vol 498, pg 355, 2013), Nature, 500, 491-491, 2013. Augustin, S., Wex, H

  8. Cutoff Values of Serum Carcinoembryonic Antigen (CEA) in Normal Korean Adults and Factors Influencing Serum CEA Level

    International Nuclear Information System (INIS)

    Kim, Jong Soon; Kim, Sun Wook; Chung, June Key; Lee, Dong Soo

    1994-01-01

    Carcinoembryonic Antigen is one of most frequently checked tumor markers in cancer management. We performed statistical analysis with serum CEA data of 2626 persons who received regular health examination and were thought to be free of active disease to determine the cutoff values of serum CEA level in normal Korean adults and to study the factors influencing serum CEA levels in normal subjects. 1) The cutoff values of serum CEA in normal Korean adults in general were 9.28 ng/ml for men, 5.90 ng/ml for women. 2) Serum CEA level was influenced by age, present smoking history, sex, and abnormal findings in chest X ray. 3) Serum CEA level had no correlation with the history of amount of alcohol consumption or obesity. 4) Cutoff values of serum CEA in normal Korean adults were tabulated according to age, sex, and smoking history. Serum CEA level was influenced by age, sex, present smoking history and abnormal findings in chest X ray and cutoff values of serum CEA were tabulated according to age, sex, and smoking history.

  9. Free Convection over a Permeable Horizontal Flat Plate Embedded in a Porous Medium with Radiation Effects and Mixed Thermal Boundary Conditions

    OpenAIRE

    Najiyah S. Khasi'ie; Roziena Khairuddin; Najihah Mohamed; Mohd Zuki Salleh; Roslinda Nazar; Ioan Pop

    2012-01-01

    Problem statement: In this study, the mathematical modeling of free convection boundary layer flow over a permeable horizontal flat plate embedded in a porous medium under mixed thermal boundary conditions and radiation effects is considered. Approach: The transformed boundary layer equations are solved numerically using the shooting method. Results: Numerical solutions are obtained for the wall temperature, the heat transfer coefficient, as well as the velocity and temperature profiles. The ...

  10. Preparation of freezing quantum state for quantum coherence

    Science.gov (United States)

    Yang, Lian-Wu; Man, Zhong-Xiao; Zhang, Ying-Jie; Han, Feng; Du, Shao-jiang; Xia, Yun-Jie

    2018-06-01

    We provide a method to prepare the freezing quantum state for quantum coherence via unitary operations. The initial product state consists of the control qubit and target qubit; when it satisfies certain conditions, the initial product state converts into the particular Bell diagonal state under the unitary operations, which have the property of freezing of quantum coherence under quantum channels. We calculate the frozen quantum coherence and corresponding quantum correlations, and find that the quantities are determined by the control qubit only when the freezing phenomena occur.

  11. Evaluation in Cellulolytic Activity of Stenotrophomonas sp. in Cellulose Nitrogen Free Mineral Medium

    International Nuclear Information System (INIS)

    Honey Thet Paing Htway; San San Yu; Zaw Ko Latt

    2011-12-01

    Three bacterial strains were isolated from rice rhizospheric soil and their nitrogen fixing activity was determined in nitrogen free mineral medium and broth with glucose and cellulose as carbon sources and they produced ammonium concentration (above 3ppm) in G-NFFMM and (2-3ppm) in C-NFMM. Moreover, their cellulolytic activity was determined by DNS mothod and strain H3 having the cellulolytic activity was selected. Then, cellulose, carboxymethyl cellulose, baggasse, pea haulm, corn stem, rice straw were used as substrates and determined its reducing sugar concentration. After detection of the cellulolytic activity, the bacteria produced the highest concentration of reducing sugar on cellulose substrate at 12 day incubation period with the reducing sugar amount of 0.12mg/ml and 0.298mg/ml on CMC substrates. In the study of argicultral wastes as substrates, the selected strain, H3, produced in the reducing sugar concentration with 0.12, 0.116,0.103 and 0.098mg/ml respectively. The selected strain was identified by biochemical characterists and 16s ribosomal DNA analysis and it was Stenotrophomonas sp.

  12. Inositol and hepatic lipidosis. II. Effect of inositol supplementation and time from parturition on serum insulin, thyroxine and triiodothyronine and their relationship to serum and liver lipids in dairy cows.

    Science.gov (United States)

    Gerloff, B J; Herdt, T H; Wells, W W; Nachreiner, R F; Emery, R S

    1986-06-01

    Percutaneous liver biopsies and blood samples were obtained from 80 dairy cows in nine Michigan herds over the peripartum period. Thirty-nine cows were fed 17 g of supplemental inositol and 41 were fed a placebo. Liver biopsies were assayed for total myoinositol and triglyceride (TG) concentrations. Blood samples were assayed for serum dextran precipitable cholesterol, nonesterified fatty acids (NEFA), insulin, thyroxine (T4), free (FT4), triiodothyronine (T3) and free T3 (FT3) concentrations. Serum concentrations of insulin and the thyroid hormones decreased near parturition, with lowest concentrations occurring in the immediate postpartum period. Concentrations of T3 correlated well with T4, and the concentrations of free thyroid hormones reflected concentrations of total thyroid hormones. The percentage of hormone in the free fraction remained constant over time. Serum insulin, T3 and T4 were negatively correlated with serum NEFA and liver TG concentrations. Thyroid hormone concentrations were positively correlated with serum dextran precipitable cholesterol concentrations. Inositol supplementation was associated with reduced circulating T3 and FT3 concentrations, but not T4 and FT4 concentrations. Changes in hormone concentrations at parturition and their relationship to liver TG and serum NEFA concentrations were consistent with a metabolic adaptation by the dairy cow to the negative energy balance of early lactation.

  13. Amniocar as a proliferative medium for mesenchymal cells

    Directory of Open Access Journals (Sweden)

    V. V. Chestkov

    2014-01-01

    Full Text Available Objectives. To develop the Amniocar nutrient medium that contains fetal calf serum (FCS and growth factors cocktail for mass cultivation of human fibroblasts. To study proliferative activity of the medium on cultures of HUVEC cells of mesenchymal origin and mesenchymal stromal cells, as well as on cell culture of human amniotic fluid.Materials and methods. Determination of the rate of accumulation of the cellular mass and cell morphology in the course of cultivation of cells of various histogenesis in the Amniocar medium and nutrient medium that contains 10 % of FCS.Results. It has been demonstrated that the Amniocar medium is prevalent as compared to the standard DMEM medium with 10 % of FCS by 2 to 5 times for cultivation of skin fibroblasts, HUVEC, and mesenchymal stem cells. The Amniocar medium increased the quantity of endothelial cells that enter mitosis and maintained the culture of HUVEC cells with prolonged passaging in vitro. Clonal cultivation of human amniotic fluid cells in the Amniocar medium secured development of colonies of both fibroblast and epithelial type.Conclusions. Proliferative Amniocar medium is efficient for mass cultivation of various cells of mesenchymal origin and can be used for diagnostic purposes in medical genetics, oncology, etc.

  14. Freezing temperature protection admixture for Portland cement concrete

    Science.gov (United States)

    1996-10-01

    A number of experimental admixtures were compared to Pozzutec 20 admixture for their ability to protect fresh concrete from freezing and for increasing the rate of cement hydration at below-freezing temperatures. The commercial accelerator and low-te...

  15. Research on strength attenuation law of concrete in freezing - thawing environment

    Science.gov (United States)

    Xiao, qianhui; Cao, zhiyuan; Li, qiang

    2018-03-01

    By rapid freezing and thawing method, the experiments of concrete have been 300 freeze-thaw cycles specimens in the water. The cubic compression strength value under different freeze-thaw cycles was measured. By analyzing the test results, the water-binder ratio of the concrete under freeze-thaw environments, fly ash and air entraining agent is selected dosage recommendations. The exponential attenuation prediction model and life prediction model of compression strength of concrete under freezing-thawing cycles considering the factors of water-binder ratio, fly ash content and air-entraining agent dosage were established. The model provides the basis for predicting the durability life of concrete under freezing-thawing environment. It also provides experimental basis and references for further research on concrete structures with antifreeze requirements.

  16. Water vapor movement in freezing aggregate base materials.

    Science.gov (United States)

    2014-06-01

    The objectives of this research were to 1) measure the extent to which water vapor movement results in : water accumulation in freezing base materials; 2) evaluate the effect of soil stabilization on water vapor movement : in freezing base materials;...

  17. Clinical evaluation of free to total prostate specific antigen ratio in serum

    International Nuclear Information System (INIS)

    Cheng Wei; Deng Shouzhen; Lin Xiangtong

    1999-01-01

    Free and total prostate specific antigen (F-PSA and T-PSA) in serum were measured with immunoradiometric assay and the F/T-PSA ratio was calculated in 175 patients with T-PSA levels in the range of 4-20 μg/L. Among them 141 patients were benign prostatic hyperplasia (BPH), 23 were untreated prostate cancer (Pca untreated) and 11 were treated prostate cancer (Pca treated). The results showed that difference in F-PSA and F/T-PSA ratio for BPH group and Pca untreated group were statistically significant (P<0.01). The effectiveness of F/T-PSA ratio for Pca (89.9%) was higher than F-PSA (54.8%). The receiver-operating characteristic (ROC) curve showed an improved diagnostic efficacy of F/T-PSA ratio compared with T-PSA for discrimination between BPH and Pca. If mean F/T-PSA ratio value + 1 SE (13.2%) was used in BPH group as discrimination limits of Pca patients, the diagnostic accuracy of BPH group and Pca untreated group were 90.8% and 82.6% respectively. Thereby F/T-PSA ratio may be useful for the differentiation between BPH and prostate cancer

  18. Association of thyroid function with human serum ghrelin and leptin levels

    International Nuclear Information System (INIS)

    Wang Jinping; Xu Hao; Wu Qiulian

    2008-01-01

    Objective: To investigate the effect of different status of thyroid function (hypothyroidism and hyperthyroidism as well as euthyroid status) on serum ghrelin and leptin levels. Methods: The levels of serum ghrelin and leptin were determined by radio immunoassay in 46 untreated subjects with hyperthyroidism, 15 hyperthyroid patients achieved a euthyroid status after radioiodine 131 I therapy, 21 cases of hypothyroidism and 18 cases of normal controls, respectively. Meanwhile, the serum levels of free triiodothyronine (FT 3 ), free thyroxine (FT 4 ) and thyroid-stimulating hormone (TSH) were measured by chemiluminescence immune assay. Results: (1) The levels of serum ghrelin in untreated hyperthyroidism were significantly lower than those in hyperthyroid patients achieved a euthyroid status (t=3.21, P 3 (r=-0.29, P 4 (r=-0.26, P< 0.05), positively correlated with serum TSH (r=0.36, P<0.05); serum leptin levels did not correlate with thyroid hormone. Conclusion: The levels of serum ghrelin were differently under different thyroid functional status and correlated with thyroid hormone, while serum leptin were not. (authors)

  19. A freeze-stable formulation for DTwP and DTaP vaccines.

    Science.gov (United States)

    Xue, Honggang; Yang, Bangling; Kristensen, Debra D; Chen, Dexiang

    2014-01-01

    Inadvertent vaccine freezing often occurs in the cold chain and may cause damage to freeze‑sensitive vaccines. Liquid vaccines that contain aluminum salt adjuvants are particularly vulnerable. Polyol cryoprotective excipients have been shown to prevent freeze damage to hepatitis B vaccine. In this study, we examined the freeze-protective effect of propylene glycol on diphtheria-tetanus-pertussis-whole-cell (DTwP) and acellular (DTaP) vaccines. Pilot lots of DTwP and DTaP formulated with 7.5% propylene glycol underwent 3 freeze-thaw treatments. The addition of propylene glycol had no impact on pH, particle size distribution, or potency of the vaccines prior to freeze-thaw treatment; the only change noted was an increase in osmolality. The potencies and the physical properties of the vaccines containing cryoprotectant were maintained after freeze-thawing and for 3 months in accelerated stability studies. The results from this study indicate that formulating vaccines with propylene glycol can protect diphtheria-tetanus-pertussis vaccines against freeze damages.

  20. Freeze-drying-induced changes in the properties of graphene oxides

    International Nuclear Information System (INIS)

    Ham, Heon; Van Khai, Tran; Gil Na, Han; Jung Kwon, Yong; Yeon Cho, Hong; Woo Kim, Hyoun; Park, No-Hyung; So, Dae Sup; Lee, Joon-Woo

    2014-01-01

    We have characterized and evaluated changes in graphene oxide (GO) induced by means of freeze-drying. In order to evaluate these changes, we investigated the effects of freeze-drying and chemical reduction processes on the structure, morphology, chemical composition, and Raman properties of GO and reduced GO. The freeze-dried GO had a pore structure, maintaining a pored morphology even after thermal annealing. The freeze-dried samples were composed of a single folded nanosheet or a few nanosheets stacked and folded. The oxygen-containing functional groups were removed not only during the freeze-drying but also during the reduction processes, with an accompanying decrease in the average size of the sp 2 carbon domain (i.e. an increase in the I D /I G value). (papers)

  1. Nanocrystals-based Macroporous Materials Synthesized by Freeze-drying Combustion

    International Nuclear Information System (INIS)

    Yan, Ruiqiang; Chen, Yu; Lin, Ye; Chen, Fanglin

    2016-01-01

    We present a novel freeze-drying combustion method for synthesis of macroporous powders with nano-network, using Sm 0.2 Ce 0.8 O 1.9 (SDC) as an example. The metal nitrate salt solution mixed with glycine is frozen to form homogeneous nitrate/glycine mixture and then freeze-dried through sublimation of ice crystals. Upon combustion of the freeze-dried mixture, SDC powders with macroporous microstructure consisting of 10–20 nm nanocrystals, high surface area and excellent sinterability are achieved. High resolution transmission electron microscopy (HRTEM) analysis indicates that nanodomains due to aggregation/segregation of dopants in the SDC powders obtained from freeze-drying combustion are much smaller than those in the SDC powders synthesized by the conventional nitrate solution combustion approach, demonstrating better elemental homogeneity and improved conductivity. Using low cost precursors and simple processing conditions, freeze-drying combustion can be a versatile method to synthesize nanocrystalline powders with excellent composition homogeneity for broad applications.

  2. The impact of hypoxemia on serum total and free prostate-specific antigen levels in patients with chronic obstructive pulmonary disease.

    Science.gov (United States)

    Ozge, Cengiz; Bozlu, Murat; Ozgur, Eylem Sercan; Tek, Mesut; Tunckiran, Ahmet; Muslu, Necati; Ilvan, Ahmet

    2015-05-01

    Prostate-specific antigen (PSA) is the most important biochemical marker in the diagnosis and follow-up of patients with prostate cancer. In recent years, a relationship between PSA levels and hypoxic conditions has been described. However, no study has investigated the PSA levels in patients with chronic obstructive pulmonary disease (COPD). The aim of the present study was to investigate the impact of hypoxemia on serum total (tPSA) and free PSA (fPSA) levels in patients with COPD. Between January 2010 and January 2014, 95 male patients who hospitalized for acute exacerbations of COPD and 80 control subjects were enrolled in the study. Serum tPSA and fPSA levels and f/tPSA ratios were determined in all patients on the first day of hospitalization (exacerbation) and 7 days after the treatment (stable state). Statistical analysis included paired t test and Mann-Whitney U test. No statistically significant differences were found between COPD and control groups with regard to the baseline characteristics, except for smoking status. The levels of serum tPSA and fPSA during exacerbation of COPD were significantly higher than the levels of the stable period (p 0.05). Hypoxemia during acute exacerbation of COPD can cause a rise in serum tPSA and fPSA levels, but f/tPSA ratio is not affected. Acute exacerbation of COPD may be added to list of the events in which PSA measurements must be interpreted with caution.

  3. RESEARCH OF MOISTURE MIGRATION DURING PARTIAL FREEZING OF GROUND BEEF

    Directory of Open Access Journals (Sweden)

    V. M. Stefanovskiy

    2016-01-01

    Full Text Available The concept of «ideal product» is proposed for the study of mass transfer during partial freezing of food products by freezing plate. The ideal product is a product, in which number of factors affecting the «real product» (meat are excluded. These factors include chemical composition of meat, quality grade of raw material (NOR, DFD, PSE, cryoscopic temperature that determines the degree of water transformation into ice, the phenomenon of osmosis, rate of freezing, etc. By using the concept of «ideal product» and its implementation in a physical experiment, it is proved that the “piston effect” causing the migration of moisture is due to frozen crust formation during partial freezing of the body. During partial freezing of the product by freezing plate, «ideal» and «real» food environment is transformed from closed system into open one with inflow of moisture to unfrozen part of the body. In the «ideal product», there is an expulsion of unfrozen moisture from freezing front, so the water appears on the body surface. Thus, the displacement of moisture increases by the same law, according to which the thickness (weight of frozen layer increases. During partial freezing of ground meat, moisture does not appear on the surface of the product, but hydrates the unfrozen part of meat. The reason of this phenomenon is the expulsion of water during formation of frozen crust and water-binding capacity of meat.

  4. Spectroscopic and molecular docking studies on the charge transfer complex of bovine serum albumin with quinone in aqueous medium and its influence on the ligand binding property of the protein

    Science.gov (United States)

    Satheshkumar, Angupillai; Elango, Kuppanagounder P.

    2014-09-01

    The spectral techniques such as UV-Vis, 1H NMR and fluorescence and electrochemical experiments have been employed to investigate the interaction between 2-methoxy-3,5,6-trichloro-1,4-benzoquinone (MQ; a water soluble quinone) and bovine serum albumin (BSA) in aqueous medium. The fluorescence of BSA was quenched by MQ via formation of a 1:1 BSA-MQ charge transfer adduct with a formation constant of 3.3 × 108 L mol-1. Based on the Forster’s theory the binding distance between them is calculated as 2.65 nm indicating high probability of binding. For the first time, influence of quinone on the binding property of various types of ligands such as aspirin, ascorbic acid, nicotinimide and sodium stearate has also been investigated. The results indicated that the strong and spontaneous binding existing between BSA and MQ, decreased the intensity of binding of these ligands with BSA. Since Tryptophan (Trp) is the basic residue present in BSA, a comparison between binding property of Trp-MQ adduct with that of BSA-MQ with these ligands has also been attempted. 1H NMR titration study indicated that the Trp forms a charge transfer complex with MQ, which reduces the interaction of Trp with the ligands. Molecular docking study supported the fact that the quinone interacts with the Trp212 unit of the BSA and the free energy change of binding (ΔG) for the BSA-MQ complex was found to be -46 kJ mol-1, which is comparable to our experimental free energy of binding (-49 kJ mol-1) obtained from fluorescence study.

  5. Temporal effects of maternal and pregnancy characteristics on serum pregnancy-associated plasma protein-A and free β-human chorionic gonadotropin at 7-14 weeks' gestation

    DEFF Research Database (Denmark)

    Ball, S; Ekelund, C; Wright, D

    2013-01-01

    OBJECTIVE: The aim of this study was to investigate gestational age-dependent effects of racial origin, smoking status and mode of conception on maternal serum levels of free β-human chorionic gonadotropin (β-hCG) and pregnancy-associated plasma protein-A (PAPP-A) at 7-14 weeks' gestation. METHODS...

  6. Levels of some trace elements in serum of esophageal cancer patients

    Energy Technology Data Exchange (ETDEWEB)

    Saeed, D M. A. [Sudan Academy of Sciences, Atomic Energy Council, Khartoum (Sudan)

    2011-01-15

    The aim of this study is to measure the concentrations of six trace elements (Zn, Se, Fe, Co, Rb, Hg) in the serum of patients with esophageal cancer who were newly diagnosed before undergoing any type of treatment. Biopsy was taken from diseased esophagus and examined histologically. A total of 43 samples of patients and 36 samples from healthy control group was collected, which were then dried by freeze drying. Then analyzed by neutron activation analysis technology, using research reactor in Syria (the miniature neutron source, MNSR) with neutron flux of 1x10''1''2 n.cm ''-''2 s''-''1 was performed. Certified reference sample from the International Agency were obtained for quality control analysis to ensure the reliability of the device. All six elements were statistically analyzed, the study proved statistical significant difference concentration of following five elements, Zn, Se, Fe, Co, Rb in the serum of patients compared to the serum samples of healthy people. The concentration of Zn and Se in patients serum were less compared to serum of healthy individuals, and the concentrations of Fe, Co, Rb in serum of patients were higher than its that the serum of healthy individuals. While the concentrations of Hg was found to be higher in serum of healthy individuals compared to patients serum, but without statistical significant. The concentration presented by means with standard deviation. The findings were compared to results obtained from literature. All five elements compatible with literature. Hg has not been investigated in such studies before. Selenium and zinc supplementation are need by cancer patient in order to enhance and increase the chances for natural immunological of cancer.(Author)

  7. Levels of some trace elements in serum of esophageal cancer patients

    International Nuclear Information System (INIS)

    Saeed, D. M. A.

    2011-01-01

    The aim of this study is to measure the concentrations of six trace elements (Zn, Se, Fe, Co, Rb, Hg) in the serum of patients with esophageal cancer who were newly diagnosed before undergoing any type of treatment. Biopsy was taken from diseased esophagus and examined histologically. A total of 43 samples of patients and 36 samples from healthy control group was collected, which were then dried by freeze drying. Then analyzed by neutron activation analysis technology, using research reactor in Syria (the miniature neutron source, MNSR) with neutron flux of 1x10''1''2 n.cm ''-''2 s''-''1 was performed. Certified reference sample from the International Agency were obtained for quality control analysis to ensure the reliability of the device. All six elements were statistically analyzed, the study proved statistical significant difference concentration of following five elements, Zn, Se, Fe, Co, Rb in the serum of patients compared to the serum samples of healthy people. The concentration of Zn and Se in patients serum were less compared to serum of healthy individuals, and the concentrations of Fe, Co, Rb in serum of patients were higher than its that the serum of healthy individuals. While the concentrations of Hg was found to be higher in serum of healthy individuals compared to patients serum, but without statistical significant. The concentration presented by means with standard deviation. The findings were compared to results obtained from literature. All five elements compatible with literature. Hg has not been investigated in such studies before. Selenium and zinc supplementation are need by cancer patient in order to enhance and increase the chances for natural immunological of cancer.(Author)

  8. Dissipative corrections to particle spectra and anisotropic flow from a saddle-point approximation to kinetic freeze out

    International Nuclear Information System (INIS)

    Lang, Christian; Borghini, Nicolas

    2014-01-01

    A significant fraction of the changes in momentum distributions induced by dissipative phenomena in the description of the fluid fireball created in ultrarelativistic heavy-ion collisions actually take place when the fluid turns into individual particles. We study these corrections in the limit of a low freeze-out temperature of the flowing medium, and we show that they mostly affect particles with a higher velocity than the fluid. For these, we derive relations between different flow harmonics, from which the functional form of the dissipative corrections could ultimately be reconstructed from experimental data. (orig.)

  9. Serum deprivation induces glucose response and intercellular coupling in human pancreatic adenocarcinoma PANC-1 cells.

    Science.gov (United States)

    Hiram-Bab, Sahar; Shapira, Yuval; Gershengorn, Marvin C; Oron, Yoram

    2012-03-01

    This study aimed to investigate whether the previously described differentiating islet-like aggregates of human pancreatic adenocarcinoma cells (PANC-1) develop glucose response and exhibit intercellular communication. Fura 2-loaded PANC-1 cells in serum-free medium were assayed for changes in cytosolic free calcium ([Ca]i) induced by depolarization, tolbutamide inhibition of K(ATP) channels, or glucose. Dye transfer, assayed by confocal microscopy or by FACS, was used to detect intercellular communication. Changes in messenger RNA (mRNA) expression of genes of interest were assessed by quantitative real-time polymerase chain reaction. Proliferation was assayed by the MTT method. Serum-deprived PANC-1 cell aggregates developed [Ca]i response to KCl, tolbutamide, or glucose. These responses were accompanied by 5-fold increase in glucokinase mRNA level and, to a lesser extent, of mRNAs for K(ATP) and L-type calcium channels, as well as increase in mRNA levels of glucagon and somatostatin. Trypsin, a proteinase-activated receptor 2 agonist previously shown to enhance aggregation, modestly improved [Ca]i response to glucose. Glucose-induced coordinated [Ca]i oscillations and dye transfer demonstrated the emergence of intercellular communication. These findings suggest that PANC-1 cells, a pancreatic adenocarcinoma cell line, can be induced to express a differentiated phenotype in which cells exhibit response to glucose and form a functional syncytium similar to those observed in pancreatic islets.

  10. Non-Toxic, Low-Freezing, Drop-In Replacement Heat Transfer Fluids

    Science.gov (United States)

    Cutbirth, J. Michael

    2012-01-01

    A non-toxic, non-flammable, low-freezing heat transfer fluid is being developed for drop-in replacement within current and future heat transfer loops currently using water or alcohol-based coolants. Numerous water-soluble compounds were down-selected and screened for toxicological, physical, chemical, compatibility, thermodynamic, and heat transfer properties. Two fluids were developed, one with a freezing point near 0 C, and one with a suppressed freezing point. Both fluids contain an additive package to improve material compatibility and microbial resistance. The optimized sub-zero solution had a freezing point of 30 C, and a freezing volume expansion of 10-percent of water. The toxicity of the solutions was experimentally determined as LD(50) greater than 5g/kg. The solutions were found to produce minimal corrosion with materials identified by NASA as potentially existing in secondary cooling loops. Thermal/hydrodynamic performance exceeded that of glycol-based fluids with comparable freezing points for temperatures Tf greater than 20 C. The additive package was demonstrated as a buffering agent to compensate for CO2 absorption, and to prevent microbial growth. The optimized solutions were determined to have physically/chemically stable shelf lives for freeze/thaw cycles and longterm test loop tests.

  11. Investigations of oocyte in vitro maturation within a mouse model.

    Science.gov (United States)

    Chin, Alexis Heng Boon; Chye, Ng Soon

    2004-02-01

    This study attempted to develop a 'less meiotically competent' murine model for oocyte in vitro maturation (IVM), which could more readily be extrapolated to human clinical assisted reproduction. Oocyte meiotic competence was drastically reduced upon shortening the standard duration of in vivo gonadotrophin stimulation from 48 h to 24 h, and by selecting only naked or partially naked germinal vesicle oocytes, instead of fully cumulus enclosed oocyte complexes. With such a less meiotically competent model, only porcine granulosa coculture significantly enhanced the oocyte maturation rate in vitro, whereas no significant enhancement was observed with macaque and murine granulosa coculture. Increased serum concentrations and the supplementation of gonadotrophins, follicular fluid and extracellular matrix gel within the culture medium did not enhance IVM under either cell-free or coculture conditions. Culture medium conditioned by porcine granulosa also enhanced the maturation rate, and this beneficial effect was not diminished upon freeze-thawing. Enhanced IVM in the presence of porcine granulosa coculture did not, however, translate into improved developmental competence, as assessed by in vitro fertilization and embryo culture to the blastocyst stage.

  12. Freeze injury to roots of southern pine seedlings in the USA | South ...

    African Journals Online (AJOL)

    ... and therefore root injury was often overlooked. Many freeze-injured seedlings died within two months of the freeze event. Since freeze injury symptoms to roots were overlooked, foresters offered various reasons (other than the freeze) for the poor seedling performance. Keywords: acclimation, frost, nursery, Pinus elliottii, ...

  13. Brown bear sperm double freezing: Effect of elapsed time and use of PureSperm(®) gradient between freeze-thaw cycles.

    Science.gov (United States)

    Alvarez-Rodríguez, Manuel; Alvarez, Mercedes; López-Urueña, Elena; Martínez-Rodriguez, Carmen; Borragan, Santiago; Anel-López, Luis; de Paz, Paulino; Anel, Luis

    2013-12-01

    The use of sexed spermatozoa has great potential to captive population management in endangered wildlife. The problem is that the sex-sorting facility is a long distance from the semen collection place and to overcome this difficulty two freeze-thaw cycles may be necessary. In this study, effects of refreezing on brown bear electroejaculated spermatozoa were analyzed. We carried out two experiments: (1) to assess the effects of the two freezing-thawing cycles on sperm quality and to analyze three different elapsed times between freezing-thawing cycles (30, 90 and 180 min), and (2) to analyze the use of PureSperm between freezing-thawing cycles to select a more motile and viable sperm subpopulation which better survived first freezing. The motility, viability and undamaged acrosomes were significantly reduced after the second thawing respect to first thawing into each elapsed time group, but the elapsed times did not significantly affect the viability and acrosome status although motility was damaged. Our results with the PureSperm gradient showed higher values of viability in freezability of select sample (pellet) respect to the rest of the groups and it also showed a significant decrease in the number of acrosome damaged. In summary, the double freezing of bear semen selected by gradient centrifugation is qualitatively efficient, and thus could be useful to carry out a sex-sorting of frozen-thawed bear spermatozoa before to send the cryopreserved sample to a biobank. Given the low recovery of spermatozoa after applying a selection gradient, further studies will be needed to increase the recovery rate without damaging of the cell quality. Copyright © 2013 Elsevier Inc. All rights reserved.

  14. Free neutron-proton analyzing power at medium energies

    International Nuclear Information System (INIS)

    Newsom, C.R.

    1980-01-01

    In recent years, increasing efforts have been made to measure the nucleon-nucleon polarization parameters. To date, no free neutron-proton spin correlated parameters have been published in the energy range 500 to 800 MeV. Existing analyzing power data is of low precision and in most cases was obtained by quasi-free proton scattering. As a first step in determining the neutron-proton scattering matrix, the free neutron-proton analyzing power has been measured at the Los Alamos Physics Facility as a function of energy and angle. The experiment was performed by scattering a neutron beam from a polarized proton target. The neutron beam was generated by scattering 800 MeV protons from a Beryllium target and using the neutrons produced at 0 degrees. The incident energy ranged from 300 MeV to 800 MeV. The energy spread of the neutron beam made it possible to measure the analyzing power at different energies simultaneously. Angular distributions were taken from 60 to 170 degrees in the center of mass system (c.m.)

  15. Polyvinyl alcohol/chitosan/montmorillonite nanocomposites preparation by freeze/thaw cycles and characterization

    Directory of Open Access Journals (Sweden)

    Părpăriţă Elena

    2014-12-01

    Full Text Available Polyvinyl alcohol (PVA and chitosan (CS based hydrogels are often chosen to obtain hydrogels as being considered non-toxic for human body. The present study aims the preparation and physical chemical characterisation of hydrogels based PVA and CS by using an environmental friendly method i.e. freeze/thaw. In this method the only parameters affecting the hydrogels’ properties is the PVA concentration in solution, time and number of cycles of freezing / thawing. Repeated freezing and thawing cycles resulted in production of a highly elastic polyvinyl alcohol hydrogel with higher degree of crystallization. Adding chitosan in polyvinyl alcohol hydrogel is giving to the newly formed material, biocompatibility and antibacterial properties due to the free amino groups of chitosan. Higher mechanical and thermal characteristics of PVA/CS based hydrogels were obtained by addition of a small amount of inorganic nanoparticles (montmorillonite clay, C30B into the matrix (i.e. 1%. Scanning electron microscopy (SEM, Fourier transform infrared spectroscopy (FT-IR, near-infrared chemical imaging spectroscopy (NI-CI, X-ray diffraction (XRD, thermogravimetric analysis (TG, swelling and rheological measurements were used to characterize the polyvinyl alcohol/chitosan/montmorillonite properties. The swelling degree increased with decreasing chitosan content in hydrogels and the variation is opposite in nanocomposites, decreasing after introducing the nanoclay. The swelling behaviour was influenced by the presence of the nanoparticles. The plasticizer effect of the nanoparticles was reflected by obtaining a more compact hydrogel network with higher mechanical and thermal properties. The proposed materials can be a promising alternative in biomedical applications

  16. DSC and TMA studies on freezing and thawing gelation of galactomannan polysaccharide

    International Nuclear Information System (INIS)

    Iijima, Mika; Hatakeyama, Tatsuko; Hatakeyama, Hyoe

    2012-01-01

    Research highlights: ► Locust bean gum forms hydrogels by freezing and thawing. ► Syneresis was observed when freezing and thawing cycle (n) increased. ► Dynamic Young's modulus increased with increasing n. ► Non-freezing water content restrained by hydrogels decreased with increasing n. ► Strong gel with densely packed network structure formed with increasing n. - Abstract: Among various kinds of polysaccharides known to form hydrogels, locust bean gum (LBG) consisting of a mannose backbone and galactose side chains has unique characteristics, since LBG forms hydrogels by freezing and thawing. In this study, effect of thermal history on gelation was investigated by differential scanning calorimetry (DSC) and thermomechanical analysis (TMA). Gel/sol ratio calculated by weighing method was found to be affected by sol concentration, freezing rate and the number of freezing and thawing cycle (n). Once LBG hydrogels are formed, they are thermally stable, although syneresis was observed when n increased. Dynamic Young's modulus (E′) of hydrogels measured by TMA in water increased with increasing n and decreasing freezing rate. Non-freezing water calculated from DSC melting peak of ice in the gel decreased with increasing n and decreasing freezing rate. Morphological observation of freeze-dried gels was carried out by scanning electron microscopy (SEM). The above results indicate that weak hydrogel having large molecular network structure transformed into strong gel with densely packed network structure by increasing n and decreasing freezing rate.

  17. Freezing and low temperature photoinhibition tolerance in cultivated potato and potato hybrids

    Directory of Open Access Journals (Sweden)

    M.M. SEPPÄNEN

    2008-12-01

    Full Text Available Four Solanum tuberosum L. cultivars (Nicola, Pito, Puikula, Timo and somatic hybrids between freezing tolerant S. commersonii and freezing sensitive S. tuberosum were evaluated for their tolerance to freezing and low temperature photoinhibition. Cellular freezing tolerance was studied using ion leakage tests and the sensitivity of the photosynthetic apparatus to freezing and high light intensity stress by measuring changes in chlorophyll fluorescence (FV/FM and oxygen evolution. Exposure to high light intensities after freezing stress increased frost injury significantly in all genotypes studied. Compared with S. tuberosum cultivars, the hybrids were more tolerant both of freezing and intense light stresses. In field experiments the mechanism of frost injury varied according to the severity of night frosts. During night frosts in 1999, the temperature inside the potato canopy was significantly higher than at ground level, and did not fall below the lethal temperature for potato cultivars (from -2.5 to -3.0°C. As a result, frost injury developed slowly, indicating that damage occurred to the photosynthetic apparatus. However, as the temperature at ground level and inside the canopy fell below -4°C, cellular freezing occurred and the canopy was rapidly destroyed. This suggests that in the field visual frost damage can follow from freezing or non-freezing temperatures accompanied with high light intensity. Therefore, in an attempt to improve low temperature tolerance in potato, it is important to increase tolerance to both freezing and chilling stresses.

  18. Freezing tolerance of ectomycorrhizal fungi in pure culture.

    Science.gov (United States)

    Lehto, Tarja; Brosinsky, Arlena; Heinonen-Tanski, Helvi; Repo, Tapani

    2008-10-01

    The ability to survive freezing and thawing is a key factor for the existence of life forms in large parts of the world. However, little is known about the freezing tolerance of mycorrhizal fungi and their role in the freezing tolerance of mycorrhizas. Threshold temperatures for the survival of these fungi have not been assessed experimentally. We grew isolates of Suillus luteus, Suillus variegatus, Laccaria laccata, and Hebeloma sp. in liquid culture at room temperature. Subsequently, we exposed samples to a series of temperatures between +5 degrees C and -48 degrees C. Relative electrolyte leakage (REL) and re-growth measurements were used to assess the damage. The REL test indicated that the lethal temperature for 50% of samples (LT(50)) was between -8.3 degrees C and -13.5 degrees C. However, in the re-growth experiment, all isolates resumed growth after exposure to -8 degrees C and higher temperatures. As many as 64% of L. laccata samples but only 11% in S. variegatus survived -48 degrees C. There was no growth of Hebeloma and S. luteus after exposure to -48 degrees C, but part of their samples survived -30 degrees C. The fungi tolerated lower temperatures than was expected on the basis of earlier studies on fine roots of ectomycorrhizal trees. The most likely freezing tolerance mechanism here is tolerance to apoplastic freezing and the concomitant intracellular dehydration with consequent concentrating of cryoprotectant substances in cells. Studying the properties of fungi in isolation promotes the understanding of the role of the different partners of the mycorrhizal symbiosis in the freezing tolerance.

  19. The bone mass density in men aged over 50 and its relation to the concentration of free and total testosterone in the blood serum

    International Nuclear Information System (INIS)

    Purzycka-Jazdon, A.; Lasek, W.; Serafin, Z.; Manysiak, S.

    2003-01-01

    As the mean length of life increases, osteoporosis affects a growing number of men and women, thus becoming an important medical and socioeconomic problem in many countries. Pathogenesis and the prevalence of the osteoporosis in women are well established, however, in men, they are still controversial. In this study, the bone mass density (BMD) of the lumbar spine was determined in 100 healthy men age 50-83, using quantitative computed tomography (QCT). Also, the total serum and free testosterone was measured. The mean BMD was 123.1I39.3 mg/cm 3 , and the values below a fracture threshold were noted in 39% of subjects. The mean concentration of total and free serum testosterone was 4.3I1.7 ng/ml and 6.2I3.7 pg/ml, respectively. There was a significant (p 3 , respectively). There was no correlation found between total testosterone and BMD. Results indicate that reduced bone mass density in males over 50 is as frequent as recently reported in females. Moreover, sex hormones seem to be related to osteoporosis development in men as well. (author)

  20. CONSIDERA TIONS OF ICE MORPHOLOGY AND DRIVING FORCES IN FREEZE CONCENTRATION

    OpenAIRE

    PETZOLD MALDONADO, GUILLERMO

    2013-01-01

    Ice rnorphology (size and shape) influence decisively in sensory appreciation, texture and quality of rnany frozen foods. Ice rnorphology is also irnportant in sorne technological processes such as freeze drying and freeze concentration, which influences the efficiency ofthese processes. The overall objective of this thesis was to increase our knowledge about the control on rnorphology of the ice phase in freezing food and related processes such as freeze concentration. Freezin...

  1. Correlation of Serum Androgens and Pituitary Hormone Levels with Serum PSA Less Than 2.5 NG/ML

    Directory of Open Access Journals (Sweden)

    Mustafa Sofikerim

    2007-01-01

    Full Text Available The aim of this clinical study was to determine whether there is a relationship between total serum testosterone, free testosterone, FSH (Follicle-Stimulating Hormone, LH (Luteinizing Hormone and serum prostate specific antigen (PSA levels. We postulated that such a correlation existed then the use of hormone specific reference ranges might enhance the usefullness of PSA concentrations <2.5 ng/mL as a marker for prostate cancer.

  2. Application of Spaceborne Scatterometer for Mapping Freeze-Thaw State in Northern Landscapes as a Measure of Ecological and Hydrological Processes

    Science.gov (United States)

    McDonald, Kyle; Kimball, John; Zimmermann, Reiner; Way, JoBea; Frolking, Steve; Running, Steve

    1999-01-01

    Landscape freeze/thaw transitions coincide with marked shifts in albedo, surface energy and mass exchange, and associated snow dynamics. Monitoring landscape freeze/thaw dynamics would improve our ability to quantify the interannual variability of boreal hydrology and river runoff/flood dynamics. The annual duration of frost-free period also bounds the period of photosynthetic activity in boreal and arctic regions thus affecting the annual carbon budget and the interannual variability of regional carbon fluxes. In this study, we use the NASA scatterometer (NSCAT) to monitor the temporal change in the radar backscatter signature across selected ecoregions of the boreal zone. We have measured vegetation tissue temperatures, soil temperature profiles, and micrometeorological parameters in situ at selected sites along a north-south transect extending across Alaska from Prudhoe Bay to the Kenai Peninsula and in Siberia near the Yenisey River. Data from these stations have been used to quantify the scatterometer's sensitivity to freeze/thaw state under a variety of terrain and landcover conditions. Analysis of the NSCAT temporal response over the 1997 spring thaw cycle shows a 3 to 5 dB change in measured backscatter that is well correlated with the landscape springtime thaw process. Having verified the instrument's capability to monitor freeze/thaw transitions, regional scale mosaicked data are applied to derive temporal series of freeze/thaw transition maps for selected circumpolar high latitude regions. These maps are applied to derive areal extent of frozen and thawed landscape and demonstrate the utility of spaceborne radar for operational monitoring of seasonal freeze-thaw dynamics and associated biophysical processes for the circumpolar high latitudes.

  3. Anti-freezing of air-cooled heat exchanger by switching off sectors

    International Nuclear Information System (INIS)

    Wang, Weijia; Kong, Yanqiang; Huang, Xianwei; Yang, Lijun; Du, Xiaoze; Yang, Yongping

    2017-01-01

    Highlights: • The anti-freezing of air-cooled heat exchanger by switching off sectors is studied. • The water side heat loads of various sectors are compared for different cases. • Anti-freezing turbine back pressure is proposed and obtained for various cases. • As wind speed increases, the energy efficiency can be clearly improved by sector off. • By switching frontal sector off, anti-freezing operation is most energy efficient. - Abstract: With the air side huge heat transfer surface, the air-cooled heat exchanger will take a serious freezing risk in cold winter. Therefore, it is of benefit to the safe operation of natural draft dry cooling system to propose the anti-freezing measures. In this work, the flow and heat transfer models of the cooling air coupling with the circulating water, are developed and numerically simulated for the anti-freezing by switching various sectors off. The local thermo-flow fields of cooling air are presented, and the water side heat loads of various sectors are compared for various cases. The anti-freezing turbine back pressure is proposed and obtained for the energy efficiency analysis. The results show that the sector switching off approach can effectively prevent the air-cooled heat exchanger from freezing and improve the energy efficiency of the cooling system, especially at high wind speeds. Moreover, with the frontal sector switching off, the most energy efficient anti-freezing operation of natural draft dry cooling system can be achieved.

  4. The elevated serum urea:creatinine ratio in canine babesiosis in South Africa is not of renal origin.

    Science.gov (United States)

    de Scally, M P; Leisewitz, A L; Lobetti, R G; Thompson, P N

    2006-12-01

    Pigmented serum, usually due to free haemoglobin and/or bilirubin, is a common finding in dogs with babesiosis, resulting in interference with all biochemical tests that rely on photochemistry. This is particularly true of urea and creatinine determinations, complicating the diagnosis of acute renal failure, which is a serious complication of babesiosis. A disproportionately raised serum urea concentration of unknown origin occurs in severely anaemic canine babesiosis patients and gives rise to an increased serum urea:creatinine ratio. The assay for cystatin-C, an excellent measure of glomerular filtration rate, is unaffected by free serum haemoglobin, and due to its different intrinsic origins, is free of influence by the metabolic derangements and organ pathology, other than renal disease, encountered in canine babesiosis. Serum cystatin-C was used to compare the concentrations of serum urea and serum creatinine in dogs with the severely anaemic form of canine babesiosis as well as a canine babesiosis-free reference group. Mean serum urea and mean serum urea:creatinine ratio were significantly elevated in the babesia-infected group relative to the reference population in this study. Mean serum creatinine and mean serum cystatin-C were within the reference ranges. Therefore an elevated urea:creatinine ratio in canine babesiosis in the presence of a normal serum creatinine concentration is considered to be caused by an elevated serum urea concentration and is most likely of non-renal origin. Serum creatinine was therefore as specific a measure of renal function as serum cystatin-C in canine babesiosis in this study. The sensitivity of serum creatinine as a measure of renal function was not established by this study. Serum urea, however, proved to be of little use compared to serum cystatin-C and serum creatinine. Serum urea should therefore not be used to diagnose renal failure in canine babesiosis.

  5. Radioimmunoassay of progesterone in unextracted serum

    International Nuclear Information System (INIS)

    Haynes, S.P.; Corcoran, J.M.; Eastman, C.J.; Doy, F.A.

    1980-01-01

    A rapid, precise radioimmunoassay for progesterone in 25 μL of unextracted serum is described. Progesterone is released from its binding protein by adding an optimal amount of cortisol, which binds to the same protein (cortisol binding globulin) as progesterone. The amount of cortisol required does not cross react with the specific progesterone antibody used. This approach considerably shortens assay time and removes a tedious and imprecise stage in the conventional assay of serum progesterone. Results correlated well (r = 0.97) with a method involving organic solvent extraction of progesterone from serum. During the two years we have used this mehod in a busy diagnostic endocrine laboratory, the between-assay precision (CV) for low-, medium-, and high-concentration quality control sera was 12, 7, and 9%, respectively. Data from participation in an independent external quality-control program verified the adequacies of the method

  6. Investigation of the process of vacuum freeze drying of bacterial concentrates for the meat industry with cryogenic freezing

    Directory of Open Access Journals (Sweden)

    V. V. Poymanov

    2016-01-01

    Full Text Available The research results of the nutritional value of the products manufactured are presented in the article. The main directions of bacterial concentrates application in the meat industry were determined. The analysis of starter cultures was given. The range of products manu-factured with bacterial concentrates was analyzed. It was shown that the introduction of innovative technologies will enable dynamic development of both large and small enterprises, which will create prerequisites for the growth of the Russian market of meat products. Economic efficiency of the studied substances treatment methods was proved. The relevance of the development of technology of pro-duction of dry bacterial concentrates with cryogenic freezing was proved. An integrated approach to the development of competitive domestic technologies and equipment for cryofreezing and sublimation dehydration by the use of granulation for the intensification of the internal heat and mass transfer, reducing specific energy consumption through the use of a combined cold supply system was pro-posed. Results of the study of the kinetics of the freezing process with the traditional method and cryofreezing are given in the paper. Rational parameters of the cryofreezing process were proposed. The optimum composition of cryoprotective medium was recommended. The research of the process of bacterial concentrate vacuum sublimation dehydration in the layer and granular form were conducted. The research confirmed that the use of the cryofreezing and granulation can increase the number of viable microorganisms in the bacterial concentrate and reduce the drying time. Rational vacuum sublimation dehydration modes were proposed. Methods of reduc-ing the defects of the processed products and improvement of the efficiency of production facilities were specified. Quality indicators of dried bacterial concentrates were given. The results obtained allow to carry out engineering calculations

  7. ASSESSMENT OF THE RESIDUAL TUMOR IN PATIENTS WITH MULTIPLE MYELOMA BASED ON THE ANALYSIS OF THE FREE LIGHT CHAINS OF IMMUNOGLOBULINS IN BLOOD SERUM

    Directory of Open Access Journals (Sweden)

    T. A. Мitina

    2013-01-01

    Full Text Available Efficiency of the multiple myeloma treatment with chemotherapy including bortezomib was assessed based on determination of the level of immunoglobulin free light chains in blood serum. The method enables estimation of changes in kinetic parameters of the residual tumor, detection of the disease course prognosis, and the choice of the optimal approach to the disease therapy.

  8. Prognostic value of the serum free light chain ratio in newly diagnosed myeloma: proposed incorporation into the international staging system.

    Science.gov (United States)

    Snozek, C L H; Katzmann, J A; Kyle, R A; Dispenzieri, A; Larson, D R; Therneau, T M; Melton, L J; Kumar, S; Greipp, P R; Clark, R J; Rajkumar, S V

    2008-10-01

    To determine if the serum free light chain (FLC) ratio has prognostic value in patients with symptomatic multiple myeloma (MM), baseline serum samples from a well-characterized cohort of 790 newly diagnosed MM patients were tested with the FLC assay. FLC ratio was calculated as kappa/lambda (reference range 0.26-1.65). On the basis of the distribution of values, a cutpoint kappa/lambda FLC ratio of 32 was chosen for further analysis. Overall survival was significantly inferior in patients with an abnormal FLC ratio of 32 (n=479) compared with those with an FLC ratio between 0.03 and 32 (n=311), with median survival of 30 versus 39 months, respectively. We incorporated abnormal FLC ratio with the International Staging System (ISS) risk factors (that is, albumin or=3.5 g/l), to create a risk stratification model with improved prognostic capabilities. Patients with 0, 1, 2 or 3 adverse risk factors had significantly different overall survival, with median survival times of 51, 39, 30 and 22 months, respectively (P<0.001). These findings suggest that the serum FLC ratio at initial diagnosis is an important predictor of prognosis in myeloma, and can be incorporated into the ISS for improved risk stratification.

  9. Influence of vascular endothelial growth factor stimulation and serum deprivation on gene activation patterns of human adipose tissue-derived stromal cells

    DEFF Research Database (Denmark)

    Tratwal, Josefine; Mathiasen, Anders Bruun; Juhl, Morten

    2015-01-01

    INTRODUCTION: Stimulation of mesenchymal stromal cells and adipose tissue-derived stromal cells (ASCs) with vascular endothelial growth factor (VEGF) has been used in multiple animal studies and clinical trials for regenerative purposes. VEGF stimulation is believed to promote angiogenesis and VEGF...... stimulation is usually performed under serum deprivation. Potential regenerative molecular mechanisms are numerous and the role of contributing factors is uncertain. The aim of the current study was to investigate the effect of in vitro serum deprivation and VEGF stimulation on gene expression patterns...... of ASCs. METHODS: Gene expressions of ASCs cultured in complete medium, ASCs cultured in serum-deprived medium and ASCs stimulated with VEGF in serum-deprived medium were compared. ASC characteristics according to criteria set by the International Society of Cellular Therapy were confirmed by flow...

  10. Reduced freezing in posttraumatic stress disorder patients while watching affective pictures

    NARCIS (Netherlands)

    Fragkaki, I.; Roelofs, K.; Stins, J.F.; Jongedijk, R.A.; Hagenaars, M.A.

    2017-01-01

    Besides fight and flight responses, animals and humans may respond to threat with freezing, an adaptive response characterized by bradycardia and physical immobility. Risk assessment is enhanced during freezing to promote optimal decision-making. Indeed, healthy participants showed freezing-like

  11. Apparatus for freeze drying of biologic and sediment samples

    International Nuclear Information System (INIS)

    Anon.

    1978-01-01

    Freeze drying to obtain water from individual samples, though not complicated, usually requires considerable effort to maintain the cold traps on a 24-hr basis. In addition, the transfer of a sample from sample containers to freeze-dry flasks is usually made with some risk of contamination to the sample. If samples are large, 300 g to 600 g, usually several days are required to dry the samples. The use of an unattended system greatly improves personnel and drying efficiency. Commercial freeze dryers are not readily applicable to the problems of collecting water from individual samples, and lab-designed collectors required sample transfer and continual replenishment of the dry ice. A freeze-dry apparatus for collecting water from individual sediment and/or biological samples was constructed to determine the tritium concentrations in fish for dose calcaluations and the tritium distribution in sediment cores for water movement studies. The freeze, dry apparatus, which can handle eight samples simultaneously and conveniently, is set up for unattended 24-hr operation and is designed to avoid sample transfer problems

  12. Lung Cancer Serum Biomarker Discovery Using Label Free LC-MS/MS

    Science.gov (United States)

    Zeng, Xuemei; Hood, Brian L.; Zhao, Ting; Conrads, Thomas P.; Sun, Mai; Gopalakrishnan, Vanathi; Grover, Himanshu; Day, Roger S.; Weissfeld, Joel L.; Wilson, David O.; Siegfried, Jill M.; Bigbee, William L.

    2011-01-01

    Introduction Lung cancer remains the leading cause of cancer-related death with poor survival due to the late stage at which lung cancer is typically diagnosed. Given the clinical burden from lung cancer, and the relatively favorable survival associated with early stage lung cancer, biomarkers for early detection of lung cancer are of important potential clinical benefit. Methods We performed a global lung cancer serum biomarker discovery study using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in a set of pooled non-small cell lung cancer (NSCLC) case sera and matched controls. Immunoaffinity subtraction was used to deplete the top most abundant serum proteins; the remaining serum proteins were subjected to trypsin digestion and analyzed in triplicate by LC-MS/MS. The tandem mass spectrum data were searched against the human proteome database and the resultant spectral counting data were used to estimate the relative abundance of proteins across the case/control serum pools. The spectral counting derived abundances of some candidate biomarker proteins were confirmed with multiple reaction monitoring MS assays. Results A list of 49 differentially abundant candidate proteins was compiled by applying a negative binomial regression model to the spectral counting data (pbiomarkers with statistically significant differential abundance across the lung cancer case/control pools which, when validated, could improve lung cancer early detection. PMID:21304412

  13. Comparison Study of Three Common Technologies for Freezing-Thawing Measurement

    Directory of Open Access Journals (Sweden)

    Xinbao Yu

    2010-01-01

    Full Text Available This paper describes a comparison study on three different technologies (i.e., thermocouple, electrical resistivity probe and Time Domain Reflectometry (TDR that are commonly used for frost measurement. Specially, the paper developed an analyses procedure to estimate the freezing-thawing status based on the dielectric properties of freezing soil. Experiments were conducted where the data of temperature, electrical resistivity, and dielectric constant were simultaneously monitored during the freezing/thawing process. The comparison uncovered the advantages and limitations of these technologies for frost measurement. The experimental results indicated that TDR measured soil dielectric constant clearly indicates the different stages of the freezing/thawing process. Analyses method was developed to determine not only the onset of freezing or thawing, but also the extent of their development. This is a major advantage of TDR over other technologies.

  14. Adaptive Control of Freeze-Form Extrusion Fabrication Processes (Preprint)

    National Research Council Canada - National Science Library

    Zhao, Xiyue; Landers, Robert G; Leu, Ming C

    2008-01-01

    Freeze-form Extrusion Fabrication (FEF) is an additive manufacturing process that extrudes high solids loading aqueous ceramic pastes in a layer-by-layer fashion below the paste freezing temperature for component fabrication...

  15. 125I Radioimmunoassay of serum ursodeoxycholyl conjugates

    International Nuclear Information System (INIS)

    Hill, A.; Ross, P.E.; Bouchier, I.A.D.

    1983-01-01

    A radioimmunoassay for serum ursodeoxycholic conjugates using an iodine-125 ligand has been developed. The bile acid was present in normal fasting serum (0.19 +- SD 0.19 μmol/l, n=24) and 2-hour post-prandial serum (0.8 +- SD 0.8 μmol/l, n=16). Gallstone patients undergoing oral ursodeoxycholic acid therapy had significantly higher post-prandial serum levels (21.5 +- SD 14.0 μmol/l, n=15) by radioimmunoassay. Gas liquid chromatography analysis indicated that in normal serum ursodeoxycholic acid was totally conjugated, whereas sera from gallstone patients contained a proportion as the free bile acid (10.2 +- SD 8.1 μmol/l, n=15). Following an oral dose of ursodeoxycholic acid, both unconjugated and conjugated forms of the bile acid appeared in the serum of healthy individuals. (Auth.)

  16. Medium effects on spin observables of proton knockout reactions

    International Nuclear Information System (INIS)

    Krein, G.; Maris, T.A.J.; Rodrigues, B.B.; Veit, E.A.

    1994-07-01

    Medium modifications of the properties of bound nucleons and mesons are investigated by means of medium energy quasi free proton knockout reactions with polarized incident protons. The sensitivity of the spin observables of these reactions to modifications of the nucleon and meson properties is studied using the Bonn one-boson exchange model of the nucleon-nucleon interaction. A method proposed to extract the pp analysing power in medium from the (p, 2 p) asymmetries indicates a reduction of this quantity compared to its free space value. This reduction is linked to modifications of masses and coupling constants of the nucleons and mesons in the nucleus. The implications of these modifications for another spin observable to be measured in the future are discussed. (author). 39 refs, 9 figs

  17. Medium effects on spin observables of proton knockout reactions

    Energy Technology Data Exchange (ETDEWEB)

    Krein, G [Instituto de Fisica Teorica (IFT), Sao Paulo, SP (Brazil); Maris, T A.J.; Rodrigues, B B; Veit, E A [Rio Grande do Sul Univ., Porto Alegre, RS (Brazil). Inst. de Fisica

    1994-07-01

    Medium modifications of the properties of bound nucleons and mesons are investigated by means of medium energy quasi free proton knockout reactions with polarized incident protons. The sensitivity of the spin observables of these reactions to modifications of the nucleon and meson properties is studied using the Bonn one-boson exchange model of the nucleon-nucleon interaction. A method proposed to extract the pp analysing power in medium from the (p, 2 p) asymmetries indicates a reduction of this quantity compared to its free space value. This reduction is linked to modifications of masses and coupling constants of the nucleons and mesons in the nucleus. The implications of these modifications for another spin observable to be measured in the future are discussed. (author). 39 refs, 9 figs.

  18. Okara: A Nutritionally Valuable By-product Able to Stabilize Lactobacillus plantarum during Freeze-drying, Spray-drying, and Storage

    Science.gov (United States)

    Quintana, Gabriel; Gerbino, Esteban; Gómez-Zavaglia, Andrea

    2017-01-01

    Okara is a nutritionally valuable by-product produced in large quantities as result of soymilk elaboration. This work proposes its use as both culture and dehydration medium during freeze-drying, spray-drying, and storage of Lactobacillus plantarum CIDCA 83114. Whole and defatted okara were employed as culture media for L. plantarum CIDCA 83114. The growth kinetics were followed by plate counting and compared with those of bacteria grown in MRS broth (control). No significant differences in plate counting were observed in the three media. The fatty acid composition of bacteria grown in whole and defatted okara showed a noticeable increase in the unsaturated/saturated (U/S) fatty acid ratio, with regard to bacteria grown in MRS. This change was mainly due to the increase in polyunsaturated fatty acids, namely C18:2. For dehydration assays, cultures in the stationary phase were neutralized and freeze-dried (with or without the addition of 250 mM sucrose) or spray-dried. Bacteria were plate counted immediately after freeze-drying or spray-drying and during storage at 4°C for 90 days. Freeze-drying in whole okara conducted to the highest bacterial recovery. Regarding storage, spray-dried bacteria previously grown in whole and defatted okara showed higher plate counts than those grown in MRS. On the contrary, freeze-dried bacteria previously grown in all the three culture media were those with the lowest plate counts. The addition of sucrose to the dehydration media improved their recovery. The higher recovery of microorganisms grown in okara after freeze-drying and spray-drying processes and during storage was ascribed to both the presence of fiber and proteins in the dehydration media, and the increase in U/S fatty acids ratio in bacterial membranes. The obtained results support for the first time the use of okara as an innovative matrix to deliver L. plantarum. Considering that okara is an agro-waste obtained in large quantities, these results represent an

  19. Serum biochemical reference intervals for free-living Sand Tiger sharks (Carcharias taurus) from east Australian waters.

    Science.gov (United States)

    Otway, Nicholas M

    2015-06-01

    Sharks are top-order predators in ocean food chains and the star attractions in aquaria worldwide. Unfortunately, blood biochemistry reference intervals (RI) have been determined for few species. The study aims to establish serum biochemical RI for free-living Sand Tiger sharks (Carcharias taurus) off eastern Australia. Thirty-seven sharks were captured and their sex, length, weight, reproductive maturity, and health status were recorded. After blood collection, serum analytes were quantified using standard analytical and statistical methods. Reference intervals, means, medians, and 90% confidence intervals were generated. Physiologic data from live and necropsied sharks were used to enhance the study results. Thirty healthy sharks were included in the study. Albumin could not be detected. With the exception of ALP activity, values were unaffected by sex, length, weight, age, and life-history stage. The means (RI) were: sodium 258 (249-267) mmol/L, potassium 5.0 (4.3-5.7) mmol/L, chloride 242 (227-257) mmol/L, inorganic phosphate 1.8 (1.7-2.0) mmol/L, total calcium 3.9 (3.3-4.4) mmol/L, magnesium 1.9 (1.6-2.2) mmol/L, glucose 2.7 (2.2-3.2) mmol/L, urea 377 (360-394) mmol/L, ALP 20 (8-31) U/L, ALT 3 U/L (no RI), AST 29 (13-45) U/L, CK 42 (5-79) U/L, total protein 30 (24-36) g/L, triglyceride 0.3 (0.1-0.6) mmol/L, cholesterol 1.4 (0.9-2.1) mmol/L, creatinine 32 μmol/L (no RI), total bilirubin 1.5 μmol/L (no RI), and osmolarity 1082 (1027-1136) mmol/L. These preliminary RI will assist with the clinical evaluation and treatment of captive and free-living Sand Tiger sharks worldwide. Studies with more animals will increase the precision of upper and lower reference limits. © 2015 American Society for Veterinary Clinical Pathology.

  20. Serum homocysteine levels in cerebrovascular accidents

    OpenAIRE

    Zongte, Zolianthanga; Shaini, L.; Debbarma, Asis; Singh, Th Bhimo; Devi, S. Bilasini; Singh, W. Gyaneshwar

    2008-01-01

    Hyperhomocysteinemia has been considered an independent risk factor in the development of stroke. The present study was undertaken to evaluate serum homocysteine levels in patients with cerebrovascular accidents among the Manipuri population and to compare with the normal cases. Ninety-three cerebrovascular accident cases admitted in the hospital were enrolled for the study and twenty-seven age and sex matched individuals free from cerebrovascular diseases were taken as control group. Serum h...

  1. "On" freezing in Parkinson's disease: resistance to visual cue walking devices.

    Science.gov (United States)

    Kompoliti, K; Goetz, C G; Leurgans, S; Morrissey, M; Siegel, I M

    2000-03-01

    To measure "on" freezing during unassisted walking (UW) and test if two devices, a modified inverted stick (MIS) and a visual laser beam stick (LBS) improved walking speed and number of "on" freezing episodes in patients with Parkinson's disease (PD). Multiple visual cues can overcome "off' freezing episodes and can be useful in improving gait function in parkinsonian patients. These devices have not been specifically tested in "on" freezing, which is unresponsive to pharmacologic manipulations. Patients with PD, motor fluctuations and freezing while "on," attempted walking on a 60-ft track with each of three walking conditions in a randomized order: UW, MIS, and LBS. Total time to complete a trial, number of freezes, and the ratio of walking time to the number of freezes were compared using Friedman's test. Twenty-eight patients with PD, mean age 67.81 years (standard deviation [SD] 7.54), mean disease duration 13.04 years (SD 7.49), and mean motor Unified Parkinson's Disease Rating Scale score "on" 32.59 (SD 10.93), participated in the study. There was a statistically significant correlation of time needed to complete a trial and number of freezes for all three conditions (Spearman correlations: UW 0.973, LBS 0.0.930, and MIS 0.842). The median number of freezes, median time to walk in each condition, and median walking time per freeze were not significantly different in pairwise comparisons of the three conditions (Friedman's test). Of the 28 subjects, six showed improvement with the MIS and six with the LBS in at least one outcome measure. Assisting devices, specifically based on visual cues, are not consistently beneficial in overcoming "on" freezing in most patients with PD. Because this is an otherwise untreatable clinical problem and because occasional subjects do respond, cautious trials of such devices under the supervision of a health professional should be conducted to identify those patients who might benefit from their long-term use.

  2. Frost heave susceptibility of saturated soil under constant rate of freezing

    Science.gov (United States)

    Ryokai, K.; Iguro, M.; Yoneyama, K.

    Introduced are the results of experiments carried out to quantitatively obtain the frost heave pressure and displacement of soil subjected to artificial freezing or freezing around in-ground liquefied natural gas storage tanks. This experiment is conducted to evaluate the frost heave susceptibility of saturated soil under overconsolidation. In other words, this experiment was carried out to obtain the relation of the over-burden pressure and freezing rate to the frost heave ratio by observing the frost heave displacement and freezing time of specimens by freezing the specimens at a constant freezing rate under a constant overburden pressure, while letting water freely flow in and out of the system. Introduced are the procedures for frost heave test required to quantitatively obtain the frost heave displacement and pressure of soil. Furthermore, the relation between the frost heave susceptibility and physical properties of soil obtained by this test is reported.

  3. Effects of Dietary Coconut Oil as a Medium-chain Fatty Acid Source on Performance, Carcass Composition and Serum Lipids in Male Broilers.

    Science.gov (United States)

    Wang, Jianhong; Wang, Xiaoxiao; Li, Juntao; Chen, Yiqiang; Yang, Wenjun; Zhang, Liying

    2015-02-01

    This study was conducted to investigate the effects of dietary coconut oil as a medium-chain fatty acid (MCFA) source on performance, carcass composition and serum lipids in male broilers. A total of 540, one-day-old, male Arbor Acres broilers were randomly allotted to 1 of 5 treatments with each treatment being applied to 6 replicates of 18 chicks. The basal diet (i.e., R0) was based on corn and soybean meal and was supplemented with 1.5% soybean oil during the starter phase (d 0 to 21) and 3.0% soybean oil during the grower phase (d 22 to 42). Four experimental diets were formulated by replacing 25%, 50%, 75%, or 100% of the soybean oil with coconut oil (i.e., R25, R50, R75, and R100). Soybean oil and coconut oil were used as sources of long-chain fatty acid and MCFA, respectively. The feeding trial showed that dietary coconut oil had no effect on weight gain, feed intake or feed conversion. On d 42, serum levels of total cholesterol, low-density lipoprotein cholesterol, and low-density lipoprotein/high-density lipoprotein cholesterol were linearly decreased as the coconut oil level increased (pcoconut oil level increased (poil in diets with coconut oil is the optimum level to reduce fat deposition and favorably affect lipid profiles without impairing performance in broilers.

  4. Gradient porous hydroxyapatite ceramics fabricated by freeze casting method

    International Nuclear Information System (INIS)

    Zuo Kaihui; Zhang Yuan; Jiang Dongliang; Zeng Yuping

    2011-01-01

    By controlling the cooling rates and the composition of slurries, the gradient porous hydroxyapatite ceramics are fabricated by the freeze casting method. According to the different cooling rate, the pores of HAP ceramics fabricated by gradient freeze casting are divided into three parts: one is lamellar pores, another is column pore and the last one is fine round pores. The laminated freeze casting is in favour of obtaining the gradient porous ceramics composed of different materials and the ceramics have unclear interfaces.

  5. Elevated Concentrations of Serum Immunoglobulin Free Light Chains in Systemic Lupus Erythematosus Patients in Relation to Disease Activity, Inflammatory Status, B Cell Activity and Epstein-Barr Virus Antibodies

    DEFF Research Database (Denmark)

    Draborg, Anette H; Lydolph, Magnus; Westergaard, Marie

    2015-01-01

    , FLCs' association with Epstein-Barr virus (EBV) antibodies was examined. METHODS: Using a nephelometric assay, κFLC and λFLC concentrations were quantified in sera from 45 SLE patients and 40 healthy controls. SLE patients with renal insufficiency were excluded in order to preclude high concentrations......OBJECTIVE: In this study, we examined the concentration of serum immunoglobulin free light chains (FLCs) in systemic lupus erythematosus (SLE) patients and investigated its association with various disease parameters in order to evaluate the role of FLCs as a potential biomarker in SLE. Furthermore...... of serum FLCs due to decreased clearance. RESULTS: Serum FLC concentrations were significantly elevated in SLE patients compared to healthy controls (pdisease activity (SLE disease activity...

  6. Use of freeze-casting in advanced burner reactor fuel design

    Energy Technology Data Exchange (ETDEWEB)

    Lang, A. L.; Yablinsky, C. A.; Allen, T. R. [Dept. of Engineering Physics, Univ. of Wisconsin Madison, 1500 Engineering Drive, Madison, WI 53711 (United States); Burger, J.; Hunger, P. M.; Wegst, U. G. K. [Thayer School of Engineering, Dartmouth College, 8000 Cummings Hall, Hanover, NH 03755 (United States)

    2012-07-01

    This paper will detail the modeling of a fast reactor with fuel pins created using a freeze-casting process. Freeze-casting is a method of creating an inert scaffold within a fuel pin. The scaffold is created using a directional solidification process and results in open porosity for emplacement of fuel, with pores ranging in size from 300 microns to 500 microns in diameter. These pores allow multiple fuel types and enrichments to be loaded into one fuel pin. Also, each pore could be filled with varying amounts of fuel to allow for the specific volume of fission gases created by that fuel type. Currently fast reactors, including advanced burner reactors (ABR's), are not economically feasible due to the high cost of operating the reactors and of reprocessing the fuel. However, if the fuel could be very precisely placed, such as within a freeze-cast scaffold, this could increase fuel performance and result in a valid design with a much lower cost per megawatt. In addition to competitive costs, freeze-cast fuel would also allow for selective breeding or burning of actinides within specific locations in fast reactors. For example, fast flux peak locations could be utilized on a minute scale to target specific actinides for transmutation. Freeze-cast fuel is extremely flexible and has great potential in a variety of applications. This paper performs initial modeling of freeze-cast fuel, with the generic fast reactor parameters for this model based on EBR-II. The core has an assumed power of 62.5 MWt. The neutronics code used was Monte Carlo N-Particle (MCNP5) transport code. Uniform pore sizes were used in increments of 100 microns. Two different freeze-cast scaffold materials were used: ceramic (MgO-ZrO{sub 2}) and steel (SS316L). Separate models were needed for each material because the freeze-cast ceramic and metal scaffolds have different structural characteristics and overall porosities. Basic criticality results were compiled for the various models

  7. [Screening serum response special antibodies of U251 cell line from surface display phage antibody library].

    Science.gov (United States)

    Yu, Min; Tan, De-Yong; Qian, Wei; Lai, Jian-Hua; Sun, Gui-Lin

    2004-05-01

    U251 cell is a sensitive cell line to serum, which stops at G0 phase of cell cycle in no-serum medium, and recovers growth when the serum is added into no-serum medium. The cell can express corresponding proteins in different phase of cell cycle. Therefore it is very signification for the study of cell cycle regulation mechanism that explores these proteins. In this paper, the mouse antibody phage display library was added into the bottle in which the serum starvation U251 cells had been cultured, and the special antibody phages were absorbed. Then the absorbed antibody phages were amplified by adding E. coli TG1 and helper phage M13K07. Amplified antibody phages were added into bottle in which the serum cultured cell after serum starvation (follow named as serum recovered cells) were incubated, so that the cell absorbed the no-special antibody phages for the serum starvation cell and the special antibody phages were in supernatant. The remaining no-special antibody phages in the supernatant were discarded by repeating above program 3-4 times. The pure special antibody phages were gotten, and amplified by adding the host cell E. coli TG1 and helper phage M13K07. Then the host bacterium infected special antibody phage was spread on the plate medium with ampicillin, and the monoclonal antibody phages were gotten. Using same as above program, the monoclonal antibody phages absorbed specially for serum recovered U251 cells were obtained when the serum recovered cells instead of serum starvation cells and serum starvation cells instead of serum recovered cells. In this study, ninety-six positive monoclonal antibody phages that absorbed specially the serum starvation cells and eighty-two positive monoclonal antibody phages that absorbed specially the serum recovered cells were obtained. By using cell immunochemistry assay, two special signification antibodies were obtained. one (No.11) was the strong response in serum starvation cells, the other (No.2) was the strong

  8. Immunomodulatory effects of supercritical fluid CO2 extracts from freeze-dried powder of Tenebrio molitor larvae (yellow mealworm

    Directory of Open Access Journals (Sweden)

    QingFeng TANG

    2016-01-01

    Full Text Available Abstract In order to take full advantage of Tenebrio molitor larvae (yellow mealworm resources, the supercritical CO2 fluid freeze-dried powder of T. molitor larvae (fdTML extraction on the immune systems of mice was carried out. The results about the effects of supercritical CO2 fluid fdTML extraction on carbon expurgation and phagocytosis of peritoneal macrophages experiments of mice indicated that the fdTML extraction enhanced observably carbon expurgatory index, phagocytic rate and phagocytic index. The fdTML extraction could stimulate response of delayed hypersensitivity. The proliferation of ConA-induced mitogenic reponse for spleen lymphocyte was also increased. The amount of hemolytic antibody in mice serum increased compared with those of the control group mice. The half of hemolysis values in serum of treated mice increased compared to the control group. Furthermore, serum NO content in all treatment groups was higher than that of the control group whereas acid phosphatase and alkaline phosphatase activity was only significantly higher relative to the control group. Our findings suggest that supercritical CO2 fluid the fdTML extraction has potential as a health food supplement.

  9. CRISPR/Cas9-Mediated Mutagenesis of Human Pluripotent Stem Cells in Defined Xeno-Free E8 Medium.

    Science.gov (United States)

    Soh, Chew-Li; Huangfu, Danwei

    2017-01-01

    The recent advent of engineered nucleases including the CRISPR/Cas9 system has greatly facilitated genome manipulation in human pluripotent stem cells (hPSCs). In addition to facilitating hPSC-based disease studies, the application of genome engineering in hPSCs has also opened up new avenues for cell replacement therapy. To improve consistency and reproducibility of hPSC-based studies, and to meet the safety and regulatory requirements for clinical translation, it is necessary to use a defined, xeno-free cell culture system. This chapter describes protocols for CRISPR/Cas9 genome editing in an inducible Cas9 hPSC-based system, using cells cultured in chemically defined, xeno-free E8 Medium on a recombinant human vitronectin substrate. We detail procedures for the design and transfection of CRISPR guide RNAs, colony selection, and the expansion and validation of clonal mutant lines, all within this fully defined culture condition. These methods may be applied to a wide range of genome-engineering applications in hPSCs, including those that utilize different types of site-specific nucleases such as zinc finger nucleases (ZFNs) and TALENs, and form a closer step towards clinical utility of these cells.

  10. Experimental research on durability of recycled aggregate concrete under freeze- thaw cycles

    Science.gov (United States)

    Cheng, Yanqiu; Shang, Xiaoyu; Zhang, Youjia

    2017-07-01

    The freeze-thaw durability of recycled aggregate concrete has significance for the concrete buildings in the cold region. In this paper, the rapid freezing and thawing cycles experience on recycle aggregate concrete was conducted to study on the effects of recycle aggregate amount, water-binder ratio and fly ash on freeze-thaw durability of recycle aggregate concrete. The results indicates that recycle aggregate amount makes the significant influence on the freeze-thaw durability. With the increase of recycled aggregates amount, the freeze-thaw resistance for recycled aggregate concrete decreases. Recycled aggregate concrete with lower water cement ratio demonstrates better performance of freeze-thaw durability. It is advised that the amount of fly ash is less than 30% for admixture of recycled aggregates in the cold region.

  11. Recent developments in novel freezing and thawing technologies applied to foods.

    Science.gov (United States)

    Wu, Xiao-Fei; Zhang, Min; Adhikari, Benu; Sun, Jincai

    2017-11-22

    This article reviews the recent developments in novel freezing and thawing technologies applied to foods. These novel technologies improve the quality of frozen and thawed foods and are energy efficient. The novel technologies applied to freezing include pulsed electric field pre-treatment, ultra-low temperature, ultra-rapid freezing, ultra-high pressure and ultrasound. The novel technologies applied to thawing include ultra-high pressure, ultrasound, high voltage electrostatic field (HVEF), and radio frequency. Ultra-low temperature and ultra-rapid freezing promote the formation and uniform distribution of small ice crystals throughout frozen foods. Ultra-high pressure and ultrasound assisted freezing are non-thermal methods and shorten the freezing time and improve product quality. Ultra-high pressure and HVEF thawing generate high heat transfer rates and accelerate the thawing process. Ultrasound and radio frequency thawing can facilitate thawing process by volumetrically generating heat within frozen foods. It is anticipated that these novel technologies will be increasingly used in food industries in the future.

  12. Measurement of free thyroxine and free triiodothyronine concentrations by DPC RIA kits

    International Nuclear Information System (INIS)

    Kasagi, Kanji; Kousaka, Tadako; Hatabu, Hiroto; Tokuda, Yasutaka; Iida, Yasuhiro; Konishi, Junji

    1988-01-01

    'DPC RIA' kits for measuring free triiodothyronine (T3) and free thyroxine (T4) were fundamentally and clinically studied. Standard curves for incubation temperature and time were favorable. The present kits yielded satisfactory precision and reproducibility. The upper and lower limits of the normal range for T4 were 0.88 ng/dl and 2.08 ng/dl, respectively. The serum concentration of T4 was 3.91 ng/dl or more for hyperthyroidism and 0.57 ng/dl or less for hypothyroidism, allowing discrimination between the two diseases. For T3, the upper and lower limits of the normal range were 1.45 ng/dl and 3.45 ng/dl, respectively. In pregnant women and patients with decreased or increased levels of thyroxine binding globulin (TBG), the serum T3 concentration lay within the normal range. Low free T4 and T3 levels were found in 22 % and 67 %, respectively, of patients with serious non-thyroidal illness (NTI). There was a good correlation between the present kits and the other commercially available kits. Measurements of free T3 and T4 obtained by ''Amerlex RIA'' kits tended to be lower than those by the present kits in pregnant women and NTI patients with hypoalbuminemia. In view of the simplified procedure, satisfactory precision and reproducibility, and the lack of influences of serum albumin and TBG, the present kits have a potential in routine use. (Namekawa, K.)

  13. Effect of freezing temperature on the color of frozen salmon.

    Science.gov (United States)

    Ottestad, Silje; Enersen, Grethe; Wold, Jens Petter

    2011-09-01

    New freezing methods developed with the purpose of improved product quality after thawing can sometimes be difficult to get accepted in the market. The reason for this is the formation of ice crystals that can give the product a temporary color loss and make it less appealing. We have here used microscopy to study ice crystal size as a function of freezing temperature by investigating the voids in the cell tissue left by the ice crystals. We have also investigated how freezing temperature affects the color and the visible absorption spectra of frozen salmon. Freezing temperatures previously determined to be the best for quality after thawing (-40 to -60 °C) were found to cause a substantial loss in perceived color intensity during frozen state. This illustrated the conflict between optimal freezing temperatures with respect to quality after thawing against visual appearance during frozen state. Low freezing temperatures gave many small ice crystals, increased light scattering and an increased absorption level for all wavelengths in the visible region. Increased astaxanthin concentration on the other hand would give higher absorption at 490 nm. The results showed a clear potential of using visible interactance spectroscopy to differentiate between poor product coloration due to lack of pigmentation and temporary color loss due to light scattering by ice crystal. This type of measurements could be a useful tool in the development of new freezing methods and to monitor ice crystal growth during frozen storage. It could also potentially be used by the industry to prove good product quality. In this article we have shown that freezing food products at intermediate to low temperatures (-40 to -80 °C) can result in paler color during frozen state, which could affect consumer acceptance. We have also presented a spectroscopic method that can separate between poor product color and temporary color loss due to freezing. © 2011 Institute of Food Technologists®

  14. Serum selenium level and risk of lung cancer mortality

    DEFF Research Database (Denmark)

    Suadicani, P; Hein, H O; Gyntelberg, F

    2011-01-01

    Serum selenium has been implicated as a risk factor for lung cancer, but the issue remains unsettled. We tested in a cohort of 3,333 males aged 53 to 74 years the hypothesis that a low serum selenium would be associated with an increased risk of lung cancer mortality.During 16 years, 167 subjects(5.......1%) died from lung cancer; 48 males (5.0%) among males with low serum selenium, 0.4-1.0 μmol·l(-1), n=965, 57 males (5.1%) among males with medium serum selenium, 1.1-1.2 μmol·l(-1), n=1,141, and 62 males (5.1%) among males with high serum selenium, 1.3-3.0 μmol·l(-1), n=1,227. After adjustment for age...... (chronic bronchitis and peak flow), referencing the lowest level of serum selenium HRs were 1.17(0.79-1.75), and 1.43(0.96-2.14), respectively. Among heavy smokers a high serum selenium was associated with a significantly increased risk of lung cancer mortality after taking into account all potential...

  15. Using a medium of free amino acids to produce penicillin g acylase in fed-batch cultivations of Bacillus megaterium ATCC 14945

    Directory of Open Access Journals (Sweden)

    R. G. Silva

    2006-03-01

    Full Text Available The production of penicillin G acylase (PGA, an important industrial enzyme from a wild strain of Bacillus megaterium using a pool of free amino acids as substrate was studied in a bench-scale bioreactor. Experiments carried out in shakers showed that the substitution of casein for free amino acids in the presence of cheese whey was the culture medium that provided the highest productivity. Several cultivations were carried out in a bioreactor operated in either batch or fed-batch mode. Batch runs showed that enzyme production is associated with microorganism growth. The following set of amino acids was preferentially consumed: Ala, Arg, Asp, Gly, Lys, Ser, Thr and Trp. On the other hand, the rates of consumption of His, Ile, Leu, Met, Phe, Pro, Tyr and Val were lower.

  16. DNA comet assay to identify different freezing temperatures of irradiated liver chicken

    International Nuclear Information System (INIS)

    Duarte, Renato C.; Mozeika, Michel A.; Fanaro, Gustavo B.; Villavicencio, Anna L.C.H.; Marchioni, Eric

    2009-01-01

    The cold chain is a succession of steps which maintain the food at low temperature. The thawed food never be frozen again and the best solution being to consume it quickly to avoid the microorganism growth which causes decay and nutrients damage. One of most important point is that freezing process, unlike irradiation, do not destroy microorganisms, only inactive them as long as they remain in a frozen state. The Comet Assay is an original test used to detect irradiated foods that's recognize the DNA damage and can then be used to control the overall degradation of the food and in a certain extend to evaluate the damage caused by irradiation, different forms of freeze and storage time on liver chicken cells. Different freezing temperatures were used, deep freeze -196 deg C and slow freeze -10 deg C. Samples were irradiated in a 60 Co irradiator with 1.5, 3.0 and 4.5 kGy radiation doses. Fast freezing technique induces a low percent of DNA degradation comparing to slow freezing technique. This procedure could be a good choose to chicken freezing processing. (author)

  17. DNA comet assay to identify different freezing temperatures of irradiated liver chicken

    Energy Technology Data Exchange (ETDEWEB)

    Duarte, Renato C.; Mozeika, Michel A.; Fanaro, Gustavo B.; Villavicencio, Anna L.C.H. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)], e-mail: renatocduarte@yahoo.com.br; Marchioni, Eric [Universite de Strasbourg, Illkirch (France). Faculte de Pharmacie. Lab. de Chimie Analytique et Sciences de l' Aliment

    2009-07-01

    The cold chain is a succession of steps which maintain the food at low temperature. The thawed food never be frozen again and the best solution being to consume it quickly to avoid the microorganism growth which causes decay and nutrients damage. One of most important point is that freezing process, unlike irradiation, do not destroy microorganisms, only inactive them as long as they remain in a frozen state. The Comet Assay is an original test used to detect irradiated foods that's recognize the DNA damage and can then be used to control the overall degradation of the food and in a certain extend to evaluate the damage caused by irradiation, different forms of freeze and storage time on liver chicken cells. Different freezing temperatures were used, deep freeze -196 deg C and slow freeze -10 deg C. Samples were irradiated in a {sup 60}Co irradiator with 1.5, 3.0 and 4.5 kGy radiation doses. Fast freezing technique induces a low percent of DNA degradation comparing to slow freezing technique. This procedure could be a good choose to chicken freezing processing. (author)

  18. Does anxiety cause freezing of gait in Parkinson's disease?

    Directory of Open Access Journals (Sweden)

    Kaylena A Ehgoetz Martens

    Full Text Available Individuals with Parkinson's disease (PD commonly experience freezing of gait under time constraints, in narrow spaces, and in the dark. One commonality between these different situations is that they may all provoke anxiety, yet anxiety has never been directly examined as a cause of FOG. In this study, virtual reality was used to induce anxiety and evaluate whether it directly causes FOG. Fourteen patients with PD and freezing of gait (Freezers and 17 PD without freezing of gait (Non-Freezers were instructed to walk in two virtual environments: (i across a plank that was located on the ground (LOW, (ii across a plank above a deep pit (HIGH. Multiple synchronized motion capture cameras updated participants' movement through the virtual environment in real-time, while their gait was recorded. Anxiety levels were evaluated after each trial using self-assessment manikins. Freezers performed the experiment on two separate occasions (in their ON and OFF state. Freezers reported higher levels of anxiety compared to Non-Freezers (p < 0.001 and all patients reported greater levels of anxiety when walking across the HIGH plank compared to the LOW (p < 0.001. Freezers experienced significantly more freezing of gait episodes (p = 0.013 and spent a significantly greater percentage of each trial frozen (p = 0.005 when crossing the HIGH plank. This finding was even more pronounced when comparing Freezers in their OFF state. Freezers also had greater step length variability in the HIGH compared to the LOW condition, while the step length variability in Non-Freezers did not change. In conclusion, this was the first study to directly compare freezing of gait in anxious and non-anxious situations. These results present strong evidence that anxiety is an important mechanism underlying freezing of gait and supports the notion that the limbic system may have a profound contribution to freezing in PD.

  19. Freeze-drying wet digital prints: An option for salvage?

    International Nuclear Information System (INIS)

    Juergens, M C; Schempp, N

    2010-01-01

    On the occasion of the collapse of the Historical Archive of the City of Cologne in March 2009 and the ensuing salvage effort, questions were raised about the use of freeze-drying for soaked digital prints, a technique that has not yet been evaluated for these materials. This study examines the effects of immersion, air-drying, drying in a blotter stack, freezing and freeze-drying on 35 samples of major digital printing processes. The samples were examined visually before, during and after testing; evaluation of the results was qualitative. Results show that some prints were already damaged by immersion alone (e.g. bleeding inks and soluble coatings) to the extent that the subsequent choice of drying method made no significant difference any more. For those samples that did survive immersion, air-drying proved to be crucial for water-sensitive prints, since any contact with the wet surface caused serious damage. Less water-sensitive prints showed no damage throughout the entire procedure, regardless of drying method. Some prints on coated media suffered from minor surface disruption up to total delamination of the surface coating due to the formation of ice crystals during shock-freezing. With few exceptions, freeze-drying did not cause additional damage to any of the prints that hadn't already been damaged by freezing. It became clear that an understanding of the process and materials is important for choosing an appropriate drying method.

  20. Intact preservation of environmental samples by freezing under an alternating magnetic field.

    Science.gov (United States)

    Morono, Yuki; Terada, Takeshi; Yamamoto, Yuhji; Xiao, Nan; Hirose, Takehiro; Sugeno, Masaya; Ohwada, Norio; Inagaki, Fumio

    2015-04-01

    The study of environmental samples requires a preservation system that stabilizes the sample structure, including cells and biomolecules. To address this fundamental issue, we tested the cell alive system (CAS)-freezing technique for subseafloor sediment core samples. In the CAS-freezing technique, an alternating magnetic field is applied during the freezing process to produce vibration of water molecules and achieve a stable, super-cooled liquid phase. Upon further cooling, the temperature decreases further, achieving a uniform freezing of sample with minimal ice crystal formation. In this study, samples were preserved using the CAS and conventional freezing techniques at 4, -20, -80 and -196 (liquid nitrogen) °C. After 6 months of storage, microbial cell counts by conventional freezing significantly decreased (down to 10.7% of initial), whereas that by CAS-freezing resulted in minimal. When Escherichia coli cells were tested under the same freezing conditions and storage for 2.5 months, CAS-frozen E. coli cells showed higher viability than the other conditions. In addition, an alternating magnetic field does not impact on the direction of remanent magnetization in sediment core samples, although slight partial demagnetization in intensity due to freezing was observed. Consequently, our data indicate that the CAS technique is highly useful for the preservation of environmental samples. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.